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WO2024069486A1 - Systèmes et appareil de bio-imprimante et procédés d'utilisation - Google Patents

Systèmes et appareil de bio-imprimante et procédés d'utilisation Download PDF

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Publication number
WO2024069486A1
WO2024069486A1 PCT/IB2023/059630 IB2023059630W WO2024069486A1 WO 2024069486 A1 WO2024069486 A1 WO 2024069486A1 IB 2023059630 W IB2023059630 W IB 2023059630W WO 2024069486 A1 WO2024069486 A1 WO 2024069486A1
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WO
WIPO (PCT)
Prior art keywords
bioink
bioprinter
cartridges
printhead
examples
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/IB2023/059630
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English (en)
Inventor
Erik PAGAN
Mohsen AKBARI
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UVic Industry Partnerships Inc
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UVic Industry Partnerships Inc
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Filing date
Publication date
Application filed by UVic Industry Partnerships Inc filed Critical UVic Industry Partnerships Inc
Publication of WO2024069486A1 publication Critical patent/WO2024069486A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A45—HAND OR TRAVELLING ARTICLES
    • A45D—HAIRDRESSING OR SHAVING EQUIPMENT; EQUIPMENT FOR COSMETICS OR COSMETIC TREATMENTS, e.g. FOR MANICURING OR PEDICURING
    • A45D34/00—Containers or accessories specially adapted for handling liquid toiletry or cosmetic substances, e.g. perfumes
    • A—HUMAN NECESSITIES
    • A45—HAND OR TRAVELLING ARTICLES
    • A45D—HAIRDRESSING OR SHAVING EQUIPMENT; EQUIPMENT FOR COSMETICS OR COSMETIC TREATMENTS, e.g. FOR MANICURING OR PEDICURING
    • A45D44/00—Other cosmetic or toiletry articles, e.g. for hairdressers' rooms
    • A45D44/005—Other cosmetic or toiletry articles, e.g. for hairdressers' rooms for selecting or displaying personal cosmetic colours or hairstyle
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/20—Apparatus for additive manufacturing; Details thereof or accessories therefor
    • B29C64/205—Means for applying layers
    • B29C64/209—Heads; Nozzles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y30/00—Apparatus for additive manufacturing; Details thereof or accessories therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y40/00—Auxiliary operations or equipment, e.g. for material handling
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y50/00—Data acquisition or data processing for additive manufacturing
    • B33Y50/02—Data acquisition or data processing for additive manufacturing for controlling or regulating additive manufacturing processes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M35/00—Devices for applying media, e.g. remedies, on the human body
    • A61M35/003—Portable hand-held applicators having means for dispensing or spreading integral media
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus

Definitions

  • the present disclosure relates to bioprinters and bioprinter systems, for example, multimaterial handheld bioprinters and automated 3D bioprinting systems.
  • In situ bioprinting the process of depositing bioinks at a defect area or site of injury, has recently emerged as a technology for tissue repair and restoration via site-specific delivery of prohealing constructs. Since its inception in the mid-1980s, three-dimensional (3D) printing has emerged as a transformative tool that can benefit and advance medicine by rapid prototyping of medical devices, implants, drug delivery systems, and complex tissues.
  • Engineering artificial tissues from cells, biomaterials, and bioactive molecules has many applications in biology and medicine. For example, engineered tissues can be used to understand disease formation and progression or to develop biological substitutes to repair or replace damaged organs, and/or to develop humanized in vitro models to validate drug safety and efficacy.
  • Bioinks are typically accomplished by a variety of techniques that include micro-extrusion-, droplet-, inkjet-, microfluidic(-assisted)-, and light-based 3D printing.
  • the structural, physical, and biological properties of bioprinted constructs can be controlled through selection of bioinks and printing method(s) to mimic the native tissue function. Further, bioprinted constructs can be used in various other applications.
  • a bioprinter device can include (i) one or more printheads (e.g., made with a high-resolution resin selective laser sintering (SLA) 3D printer), (ii) syringe cartridges adapted for hydraulic or pneumatic extrusion, (iii) a cartridge enclosure or housing, and/or (iv) a mountable photocrosslinking system comprising a case with an array of light sources that emit a selected wavelength (e.g., 3mm 405nm LEDs (light emitting diodes)). Further, the bioprinter device can be coupled to (e.g., in fluid communication with) an array of external syringe pumps for hydraulic or pneumatic material extrusion.
  • SLA high-resolution resin selective laser sintering
  • the bioprinter device and/or the array of external syringe pumps can be in data communication with and controlled by a computerized controller, which may receive feedback data or signals from a detection apparatus and adjust operation of the bioprinter device and/or the array of external syringe pumps in response thereto or based at least in part thereon.
  • the bioprinter device is a component of a 3D bioprinting system.
  • handheld multi-material 3D printers and systems can include (i) one or more printheads (e.g., made with a high-resolution resin SLA 3D printer), (ii) syringe cartridges adapted for hydraulic or pneumatic extrusion, and/or (iii) a cartridge enclosure or housing, which can be used for depositing multiple materials in various applications (such as, for example, artistic and/or esthetic applications).
  • printheads e.g., made with a high-resolution resin SLA 3D printer
  • syringe cartridges adapted for hydraulic or pneumatic extrusion e.g., a high-resolution resin SLA 3D printer
  • a cartridge enclosure or housing which can be used for depositing multiple materials in various applications (such as, for example, artistic and/or esthetic applications).
  • an assembly for adapting a 3D printer to a bioprinter or other multi-material 3D printer can include (i) one or more printheads (e.g., made with a high-resolution resin SLA 3D printer), (ii) syringe cartridges adapted for hydraulic or pneumatic extrusion, (iii) a cartridge enclosure or housing, (iv) a mountable photocrosslinking system comprising a case with an array of light sources that emit a selected wavelength (e.g., 3mm 405nm LEDs (light emitting diodes)), (v) a controller (e.g., a touch sensitive GUI, a processor, and/or a memory), and/or (vi) a computer readable storage media having computer-readable instructions (e.g., programs) stored thereon for controlling operation of the 3D bioprinter and/or other firmware and applications for controlling operation of the 3D bioprinter.
  • one or more printheads e.g., made with a high-resolution resin SLA
  • a bioprinting assembly can include a multi-axis robotic arm and a bioprinter attachable thereto.
  • the bioprinting assembly can be utilized in surgical procedures.
  • the surgical assembly can include an imaging system for providing real-time feedback to a computerized controller, which can control motions of the robotic arm based thereon.
  • the handheld bioprinters and bioprinting systems described herein have advantages over known bioprinting devices.
  • combined stereolithography (SLA) 3D printing and microfluidic technologies can enable the handheld bioprinters to have relatively low manufacturing and operating costs relative to known bioprinters.
  • the ergonomic design of the handheld bioprinters can facilitate the shape-controlled biofabrication of multi-component fibers with different cross-sectional shapes and material compositions.
  • the handheld bioprinters and bioprinting systems can be used in applications where the printed fibers are utilized for on-demand, temporal, and dosage-control drug delivery platforms.
  • the handheld bioprinters and bioprinting systems can be used in applications for generating biosensors and wearable electronics via incorporating conductive materials and integrating pH responsive dyes.
  • the handheld bioprinters and bioprinting systems can be used in applications to generate cell-laden fibers with high cell viability for site-specific cell delivery by, for example, producing single component and multi-component cell-laden fibers.
  • the handheld bioprinters and bioprinting systems can be used to generate multicomponent fibers that model the invasion of cancer cells into the adjacent tissue.
  • FIGS. 1A-1C are respectively elevation, top perspective, and bottom perspective views of an exemplary handheld bioprinter, in accordance with the present disclosure.
  • FIGS. 2 A and 2B are respectively isometric and elevation views of the handheld bioprinter of FIGS. 1 A-1C with a housing and a cooling or temperature control module of the bioprinter removed.
  • FIG. 2C is an elevation view of the handheld bioprinter of FIGS. 1 A-1C with a cooling or temperature control module of the bioprinter removed.
  • FIGS. 3 and 4 are respectively perspective views of first and second cooling or temperature control module for use with a handheld bioprinter.
  • FIG. 5 is a bottom perspective view of a light curing module for use a handheld bioprinter.
  • FIGS. 6 A and 6B are respectively top and bottom perspective views of the light curing module of FIG. 5 mounted on a handheld bioprinter.
  • FIG. 7 is an exploded view of a bioprinter assembly including the handheld bioprinter of FIGS. 1 A-1C, the cooling or temperature control module of FIG. 4, and the light curing module of FIG. 5.
  • FIG. 8 is a cross-section of the bioprinter assembly of FIG. 7.
  • FIG. 9 is a perspective view illustrating exemplary operation of the bioprinter assembly of FIG. 7.
  • FIG. 10 is a perspective view of an exemplary handheld bioprinter system.
  • FIGS. 11 A-l II are transparent perspective and detailed views of exemplary printheads that can be utilized with a handheld bioprinter and their corresponding print output configurations.
  • FIG. 12A is transparent perspective and detailed view of an exemplary single channel printhead and exemplary channel shape and its corresponding print output configuration.
  • FIGS. 12B-12I are transparent perspective views of additional exemplary printheads that can be utilized with a handheld bioprinter and their corresponding print output configurations.
  • FIG. 13Ai-xii are cross-sectional microscopic images of fibers produced with the various ones of the exemplary printheads of FIGS. 11 A-l II and 12A-12I.
  • FIGS. 13Axiii-xv are photographic images of exemplary fibers that can be produced with the handheld bioprinter system.
  • FIG. 13Bi-v are photographic images of bioprinted meshes on ex vivo pig skins with various wound shapes utilizing the handheld bioprinter system.
  • FIG. 13C includes photographic images of handheld bioprinter printing a multi-material construct directly on an artificially made pig skin wound utilizing the handheld bioprinter system.
  • FIGS. 14A-14D are graphs showing time-release properties of fibers generated with the handheld bioprinter system.
  • FIGS. 15A is a cross-sectional view and a detailed transparent perspective view of a dualchannel printhead for use with a bioprinting system disclosed herein.
  • FIGS. 15B-15C are schematic illustrations and microscopic images of fibers generated with the dual-channel printhead of FIG. 15 A.
  • FIGS. 16A-16B are photographic images of exemplary fibers that can be produced with a bioprinting system disclosed herein.
  • FIGS. 16C-16H are microscopic images of exemplary fibers that can be produced with a bioprinting system disclosed herein.
  • FIG. 161 is a graph illustrating relative fluorescence intensity of the cell- loaded GelMA meshes after 1 and 2 days of culturing.
  • FIGS. 17A-17B are transparent perspective views of exemplary printheads and bioink materials for use with a bioprinting system disclosed herein.
  • FIGS. 17C-17E are microscopic images of exemplary fibers that can be produced with the printhead of FIG. 17A.
  • FIGS. 17F-17H are microscopic images of exemplary fibers that can be produced with the printhead of FIG. 17B.
  • FIG. 18 A is a cross-sectional view and a detailed transparent perspective view of a dualcore printhead for use with a bioprinting system disclosed herein.
  • FIGS. 18B-18C are schematic illustrations and microscopic images of fibers generated with the dual-channel printhead of FIG. 18 A.
  • FIG. 18D is a photographic image of exemplary fibers that can be generated with the dualchannel printhead of FIG. 18 A.
  • FIGS. 18E-18F are a graph of relative fluorescence intensity and a graph of invasion length of mono-cultured and co-cultured GelMA-alginate in dual-core fibers.
  • FIG. 19A is a cross-sectional view and a detailed transparent perspective view of a coaxial printhead for use with a bioprinting system disclosed herein.
  • FIGS. 19B-19D are each a schematic view and a corresponding photographic image of exemplary fibers that can be generated with the coaxial printhead of FIG. 19 A.
  • FIGS. 19E-19F are photographic images of exemplary fibers that can be generated with the coaxial printhead of FIG. 19A.
  • FIG. 19G is a microscopic image of an exemplary fiber that can be generated with the coaxial printhead of FIG. 19A.
  • FIGS. 20A-20D are photographic images of exemplary fibers that can be produced with the handheld bioprinter system.
  • FIGS. 21 and 22 are functional block diagrams of first and second exemplary computerized bioprinter systems, respectively.
  • FIG. 23 is a table of features of the bioprinters and bioprinter systems disclosed herein and features of existing bioprinters.
  • FIG. 24 is an elevation view of an exemplary hydraulic cartridge, in accordance with the present disclosure.
  • FIGS. 25A-25B are transparent perspective view of an exemplary mixing printhead for use with the bioprinter and/or bioprinting systems disclosed herein.
  • FIG. 25C is a transparent elevation view of the mixing printhead of FIGS. 25A-25B.
  • FIG. 26 is an elevation view of a bioprinter system including the mixing printhead of FIGS. 25A-25C.
  • FIG. 27A-27B are perspective views of an exemplary 3D -printer bioprinting system.
  • FIG. 27C is a perspective detail view of an exemplary printhead and its output for the 3D- printer bioprinting system of FIGS. 27A-27B.
  • FIG. 28A is an elevation view of an exemplary carriage for a bioprinter for use with the 3D-printer bioprinting system of FIGS. 27A-27B.
  • FIG. 28B is an elevation view of the exemplary carriage of FIG. 28A with an outer housing thereof removed for illustrative purposes.
  • FIG. 29A is a perspective view of a carriage assembly including the carriage of FIGS. 28A- 28B.
  • FIGS. 29B-29C are exploded views of the carriage assembly of FIG. 29A.
  • FIG. 30 is a schematic illustration of a robotic bioprinter system including a functional block diagram of computerized systems in communication with one or more components of the robotic bioprinter system.
  • values, procedures, or apparatus may be referred to as “lowest,” “best,” “minimum,” or the like. It will be appreciated that such descriptions are intended to indicate that a selection among many alternatives can be made, and such selections need not be better, smaller, or otherwise preferable to other selections.
  • 3D printing has emerged as a transformative tool that can benefit and advance medicine by rapid prototyping of medical devices, implants, drug delivery systems, and complex tissues.
  • Engineering artificial tissues from cells, biomaterials, and bioactive molecules has many applications in biology and medicine. These engineered tissues are used to understand disease formation and progression or to develop biological substitutes to repair or replace damaged organs. Lab-grown tissues can also be utilized to develop humanized in vitro models to validate drug safety and efficacy.
  • bioinks are materials that can be used to produce engineered/artificial live tissue using 3D printing, and can include cells suspended in a matrix such as any of various biopolymer gels.
  • cell and bioink deposition can typically be achieved via robotic arms or handheld printers that deliver materials onto the surface.
  • the body itself acts as a bioreactor to mature the tissue/organ.
  • no further manipulation of the final construct is needed.
  • bioprinting technologies have been evolving rapidly due to their inherent advantages over both conventional and in situ bioprinting.
  • This approach enables direct control over biomaterial deposition, eliminating the requirement for medical imaging to generate a 3D model of the implant. This significantly reduces the amount of equipment required during use and removes the limitations associated with computer-aided control for printing constructs directly on the wound site.
  • Accurate deposition of bioinks is typically accomplished by a variety of techniques that include micro-extrusion-, droplet-, inkjet-, microfluidic(-assisted)-, and light-based 3D printing.
  • microfibers generated by existing handheld bioprinters and biofabrication technologies are limited to either homogeneous, or simple dual-material chemical compositions and rudimental fiber morphologies (for example, microfibers including only two colinear or coaxial bioinks), and therefore are not able to mimic the composition of complex human tissues.
  • existing handheld bioprinters are only compatible with a narrow range of biomaterials (for example, being compatible with only photocrosslinkable bioinks), and the devices often require a rigorous fabrication process, all of which strongly restrict their range of applications.
  • some known bioprinting devices are only capable of generating sheets with a consistent thickness and width, and such sheet architecture may fail to entirely cover the edges and wound area.
  • the pressure in known bioprinters may require constant adjustment in order to maintain a constant flowrate due to variations in temperature caused by the heat from the user’ s hand and/or ambient heat while operating the device. Further still, such changes in temperature can undesirably decrease or increase viscosity of a bioink, and thereby limit a period of use of an individual bioink cartridge. Still further, as conventional bioprinters can use only one or two bioinks, the user is required to change bioink cartridges out in order to switch to different bioinks, thereby extending a total time required for bioink deposition.
  • known bioprinters often include a stepper motor, syringe pumps, batteries, and/or other electrical and mechanical components enclosed in the device that add a significant amount of size and weight or bulk to a handheld device.
  • the size and weight of known bioprinters negatively impacts ergonomics and comfort in operation of the device, thereby hindering long-term operation of the bioprinting device. Further, such devices are expensive to manufacture or fabricate.
  • the size and weight of known bioprinters limits a number of bioink cartridges that can be simultaneously received within and/or operatively connect to a handheld bioprinter. For example, each bioink cartridge requires an onboard stepper motor, and therefore existing handheld bioprinters typically have a maximum of two stepper motors (and are thus configured to receive a maximum of two bioink cartridges) in order to maintain single-handed operability of the device.
  • bioprinter devices that enable structural, physical, and biological properties of bioprinted constructs to be generated through selection of multiple bioinks, selection of configurations and patterns, and/or control of spatial-temporal distribution to mimic the native tissue function. Indeed, recapitulating the multiscale complexities of tissues can benefit from improved technologies that involve high resolution printing of multiple bioinks, creating sophisticated multi-luminal structures.
  • bioprinter devices and bioprinting systems disclosed herein address one or more of the foregoing issues with conventional bioprinters to meet the need for improved bioprinting of tissues.
  • the handheld bioprinters disclosed herein can be capable of depositing multiple materials with control over their spatiotemporal physicochemical properties.
  • the bioprinters disclosed herein can be modular and can be equipped with a mountable lightemitting module that facilitates photo-crosslinking while printing photocrosslinkable bioinks.
  • the handheld bioprinters disclosed herein can include a passive temperature control module that substantially maintains a selected temperature (e.g., maintains a constant temperature) of bioink cartridges for extended periods, allowing for long-term printing of viscous, thermally gelled, and/or cell-loaded bioinks.
  • a passive temperature control module that substantially maintains a selected temperature (e.g., maintains a constant temperature) of bioink cartridges for extended periods, allowing for long-term printing of viscous, thermally gelled, and/or cell-loaded bioinks.
  • an off-device pump system enables a lower cost, improved ergonomic hand piece that can hold three or more bioinks simultaneously (or fewer bioinks if desired).
  • the bioprinters disclosed herein enable multi-material printing via use of multi-channel printheads with complex fluidic circuitry that enable creating complex flows of several bioinks in microscale.
  • the versatility afforded by the modular assembly and wide range of configurations of the rapid-prototyped printheads can allow for fabrication of microfibers with diverse compositions and geometries.
  • the bioprinters disclosed herein can be used in applications for printing multiple cells, drug-releasing meshes, and/or biosensors.
  • a bioprinter device 10 can include (i) a printhead 12 made with, for example, a high-resolution resin SLA 3D printer, (ii) pneumatic syringe cartridges 14 adapted for material extrusion, (iii) a 3D printed cartridge enclosure or housing 16, (iv) a mountable photocrosslinking system 18 (also referred to herein as a “light curing module”), which can comprise a 3D printed case with an array of inexpensive 3mm 405nm LEDs (light emitting diodes), and/or (v) a cooling or temperature control module 20.
  • a printhead 12 made with, for example, a high-resolution resin SLA 3D printer
  • pneumatic syringe cartridges 14 adapted for material extrusion
  • a 3D printed cartridge enclosure or housing 16
  • a mountable photocrosslinking system 18 also referred to herein as a “light curing module”
  • a cooling or temperature control module 20 As can be seen in FIG.
  • the bioprinter device can be coupled to (e.g., in fluid communication with) an array of external syringe pumps 26 for hydraulic or pneumatic bioink material extrusion.
  • the bioprinter 10 can be utilized with printheads 12 having different (various) configurations.
  • the bioprinters disclosed herein can be used in a variety of applications.
  • the bioprinters disclosed herein can be a component of a bioprinting system 100, 200.
  • Having the syringe pumps separate from the handpiece or bioprinter device can reduce its size and weight compared to direct motor driven extrusion mechanisms, resulting in a more ergonomic and comfortable design for improved long-term operation, while allowing for the addition of multiple materials (e.g., three or more bioinks cartridges housed within the handpiece) and maintaining a compact form factor.
  • multiple materials e.g., three or more bioinks cartridges housed within the handpiece
  • a hydraulic-driven extrusion mechanism or system can have advantages over conventional handheld extrusion mechanisms, such as pressure driven extrusion, since the flowrate can be directly set on the syringe pump and can remain constant regardless of the operating viscosity and temperature of a particular bioink, eliminating the need to adjust pressure values for different types of bioinks and temperature variations, which can be an arduous process, especially for low viscosity materials.
  • the pump array can include a pneumatic pump mechanism or system.
  • bioinks are materials used to produce engineered/artificial live tissue using 3D printing.
  • Exemplary bioinks that can be utilized with the bioprinters described herein include sterile GelMA bioink, alginate-laponite bioink, PEGDA bioink, PEGDA-laponite bioink, sterile alginate-GelMA bioink, pH-responsive colorimetric bioink, conductive graphene oxide (GO) bioink, cell-laden bioink, and/or other bioinks, chemical additives, and/or combinations of the foregoing materials.
  • the bioprinters described herein can be used to print/extrude any of a variety of other materials in a flowable state, including liquids such as aqueous solutions, gels, hydrogels, or other flowable materials.
  • the bioprinters disclosed herein can be a component of a programmable bioprinting system, where a computerized controller can be configured to control bioink deposition from the bioprinter for bioink deposition in specified protocols, recipes, spatial and/or temporal patterns, etc.
  • the system can include a detection apparatus configured to provide feedback signals or data to the computerized controller for adjusting bioink deposition (if necessary) to meet one or more predefined criteria or other parameters for deposition.
  • the bioprinters disclosed herein can be a component of an automated 3D bioprinter.
  • one or more of the bioprinter devices or extruders can each be mounted onto single-axis or multi-axis positioning systems, such as robotic arms (or other robotic positioning control systems), for control of positioning and movement of the respective bioprinter device.
  • the bioprinter device can be stationary and the deposition area can robotically moveable relative to the bioprinter device.
  • both the bioprinter device and the deposition area or surface can include a robotic mechanism for controlling movement and positioning thereof.
  • the bioprinter devices and the robotic positioning control mechanisms can be controlled by a computerized controller for bioink deposition in specified protocols, recipes, spatial and/or temporal patterns, etc.
  • the automated 3D printer can include a detection device or sensor configured to provide feedback signals or data to the computerized controller for adjusting bioink deposition to meet one or more predefined criteria or other parameters for deposition.
  • FIG. 23 includes a table illustrating exemplary differentiating features of the bioprinter apparatus and systems disclosed herein relative to known bioprinter devices.
  • FIGS. 24-29C illustrate additional exemplary bioprinter components and bioprinter systems and assemblies in accordance with the present disclosure.
  • the handheld bioprinter 10 can include a multi-channel printhead 12 (e.g., a three or more channel printhead) which may be interchangeable with other printheads, the light curing module 18 (also referred to as a “LED module”) attached thereto (a mountable photocrosslinking system which may be releasable from the printhead), and a cooling or temperature control module 20.
  • a multi-channel printhead 12 e.g., a three or more channel printhead
  • the light curing module 18 also referred to as a “LED module”
  • a mountable photocrosslinking system which may be releasable from the printhead
  • a cooling or temperature control module 20 e.g., a cooling or temperature control module
  • bioink cartridges 14 e.g., two or more bioink cartridges, such as, three or four bioink cartridges
  • a housing 16 also referred to as an “enclosure” (which can be a separate enclosure or can be a central channel of the cooling module) and fluidly coupled to the multi-channel printhead 12 (FIG. 2C).
  • each of the bioink cartridges 14 includes fluid coupling and sealing mechanisms 22a, 22b at each end of the cartridge.
  • the sealing mechanism 22a can be a first Luer lock adapter, and each cartridge 14 can be coupled to (e.g., have inserted therein) the first Luer lock adapter at a first end thereof (inflow end) for fluidly coupling the cartridge 14 to a tube 24 that brings the cartridge into communication with the respective hydraulic syringe pump in the hydraulic pump array 26 (FIGS. 9 and 10).
  • Each of the Luer locks coupled at the first end of the cartridge 14 can include a female-to-barb adapter, where the barb is configured to be coupled to the hydraulic pump tube 24, a push-fit male Luer lock adapter, and an O-ring configured to form a seal between the push-fit male Luer lock adapter and an interior wall of the bioink cartridge 14 (FIG. 7).
  • the sealing mechanism 22b can be a second Luer lock adapter, and each cartridge 14 can be further coupled to (e.g., have inserted therein) the second Luer lock adapter (e.g., a female-to-female Luer lock adapter) at a second end thereof (outflow end) for coupling the cartridge 14 to a respective channel 28 within the multi-channel printhead 12.
  • alternate or additional coupling components, mechanisms, and/or structures can be utilized for fluidly coupling the cartridges to the printhead 12 and/or the hydraulic pump array 26.
  • the printhead 12 can include a base portion 30 comprising an inflow interface configured to enable coupling of bioink cartridges 14 thereto (for example, configured to enable three or more bioink cartridges thereto), a tip portion 32 having a distal opening 34 configured for output of bioink, and bioink channels 28 (for example, three or more bioink channels) that extend through the base 30 and the tip portion 32 to enable flow of bioink from the bioink cartridges through the base and the tip portion to the opening 34.
  • a diameter of a bioink channel 28 is in a range of 10 - 1000 pm, such as 10 - 500 pm, 100 - 500 pm, 10 - 100 pm, etc.
  • the cartridge 14 can be sealed between the hydraulic pump 26 and the printhead inflow interface at the base 30.
  • a piston 36 can be disposed within each bioink cartridge 14 upstream relative to the bioink material 38 disposed therein and can be acted upon by the hydraulic pump 26 to control pressure and/or flow of the bioink dispensed from the cartridge 14.
  • the bioink material 38 can flow from the cartridge 14 through the bioink channel 28 and the opening 34 in the tip 32 of the printhead 12 for deposition of bioink material 38 onto a target location (e.g., a wound or other bioink deposition area or surface 40, such as that illustrated in FIG. 9).
  • a target location e.g., a wound or other bioink deposition area or surface 40, such as that illustrated in FIG. 9.
  • flow of bioink from each cartridge can be independently controlled such that, for example, a user can dispense two or more of the bioinks simultaneously (e.g., a first bioink and a second bioink can be dispensed together) or the user can dispense two or more bioinks in sequence (e.g., dispense a first bioink and then a second bioink).
  • the bioprinter can include one or more actuators for controlling bioink flow form one or more of the bioink cartridges.
  • FIGS. 11A-11I and 12A-12I Various configurations for the printhead and bioink depositions are discussed further below with reference to FIGS. 11A-11I and 12A-12I.
  • the cooling or temperature control module 20 can include a central channel 42 that is configured to receive bioink cartridges 14.
  • the temperature control module 20 is configured to receive four bioink cartridges 14 and the central channel 42 has a 4-leaf clover or four- compartment or section configuration, where each compartment or section is configured to receive one of the bioink cartridges 14.
  • the temperature control module can be adapted to receive a different number of cartridges, such as one, two, three, five or six or more cartridges.
  • FIG. 3 illustrates a temperature control module 20’ including a central channel 42’ configured to receive one bioink cartridge 14.
  • the bioink cartridges 14 can be directly received within the temperature control module 20 (or 20’). In other examples, the bioink cartridges can be disposed within the handpiece enclosure or housing 16, and the handpiece enclosure 16 can be received within the central channel 42 (or 42’) of the temperature control module 20 (or 20’).
  • the temperature control modules 20, 20’ can be passive temperature control sleeves for preventing or limiting heating of the bioink contained within the cartridge over a specified period of time (such as, for example, maintaining a temperature in a range of 15 - 60 °C for a period of time in a range of 30 min - 1 hour). In some examples, the temperature control modules 20, 20’ can maintain a constant temperature for the period of time. Thus, the temperature control modules 20, 20’ can enable for longer-term printing of viscose and/or temperature sensitive bioinks relative to conventional bioprinters.
  • the temperature control modules 20, 20’ can include an interior wall 44, 44’ (defining the central channel), an exterior wall 46, 46’, and one or more chambers 48, 48’ disposed between the interior wall and the exterior wall.
  • the one or more chambers 48, 48’ can each be configured to receive a phase-change material that limits heating or maintains a temperature of the bioink contained within a cartridge for the period of time. It will be appreciated that different phase-change materials can maintain a bioink at a different temperature, depending on properties of the phase-change material. Accordingly, a phase-change material can be selected to maintain a specific bioink at a temperature that enables deposition of the bioink via the handheld bioprinter at a selected viscosity.
  • phase-change materials that can be used with the temperature control module (and associated temperatures) include: H2O (0°C), LiCICh-SEbO (8.1°C), ZnChGFEO (10°C), K2HPO4 6H2O (13°C), NaOH-3.5H 2 O (15.4°C), DMS (16.5°C), Polyglycol E400 (8°C), Paraffin C15-C16 (8°C), Paraffin Ci 6 -Ci 8 (20°C-22°C), Capric-lauric acid+pentadecane (90:10) (13.3°C), Isopropyl stearate (14°C-18°C), Caprylic acid (16.3°C), and Zn(NC>3)2-6H2O (36°C).
  • Exemplary commercially available phase-change materials that can be used with the temperature control module (and associated temperatures) include: ClimSel C-7 (7°C), ClimSel C-10 (10°C), ClimSel C-18 (18°C), ClimSel C-21 (21°C), RUBITHERM® RT4 (4°C), RUBITHERM RT5 (5°C), RUBITHERM RT8 (8°C), RUBITHERM RT10 (10°C), RUBITHERM RT12 (12°C), RUBITHERM RT15 (15°C), RUBITHERM RT35 (35°C), and RUBITHERM RT42 (42°C).
  • each section of the chamber that corresponds to one of the four compartments can be separated by barrier extending between the interior wall 44 and the exterior wall 46.
  • each chamber can include a phase-change material that is selected for a specific bioink.
  • the temperature control module 20 can include four different phase-change materials, each compartment having one phase-change material disposed therein that is selected for maintaining a temperature (limiting heating) for a period of time of the specific type of bioink received in an adjacent portion or compartment of the central channel.
  • the handheld bioprinters disclosed herein can maintain the temperature of bioink cartridges for extended periods, allowing for longer- term printing of viscose bioinks relative to known bioprinter devices.
  • the light curing module 18 can include one or a plurality of light sources 50 configured to emit light at a selected wavelength for photo-crosslinking of photocrosslinkable bioinks as they are being dispensed from the handheld bioprinter 10.
  • the handpiece enclosure 16 can be opaque to selected wavelengths of light (e.g., ultraviolet light) and can be configured to block or limit transmission of light therethrough in order to prevent photocrosslinking of bioink contained within a bioink cartridge mounted to the printhead.
  • the handpiece enclosure 16 can be made from PLA material on an ULTIMAKER® 2 3D printer.
  • FIGS. 6A and 6B show the light curing module 18 used in combination with a single bioink cartridge 14 having a printhead 12 coupled thereto (for example, a needle or single-channel). Additional examples are shown in FIGS. 7-8, where the light curing module 18 is used in combination with three or more bioink cartridges 14 coupled to a printhead 12 (for example, a multi-channel printhead).
  • the light curing module 18 includes a housing 52 having a central channel 54 for receiving the printhead 12 therethrough.
  • the light curing module housing 52 can be made from PLA material on an ULTIMAKER 2 3D printer.
  • a distal face 56 of the light curing module 18 can be proximate to the opening in the tip 34 of the printhead 12.
  • the light or a light array 50 e.g., an LED light array including 3mm 405nm LEDs (light emitting diodes)
  • the bulbs of the light array 50 can be radially arranged on the distal face 56.
  • the bulbs of the light array 50 can be angled inward to direct the light toward a bioink being deposited form the opening in the tip 34 of the printhead 12.
  • the light curing module 18 can include an actuator 58 for activating the light array 50.
  • the light curing module 18 can be removable from the printhead 12 when not needed. For example, when no photocrosslinkable bioink are being used in the handheld bioprinter 10.
  • FIGS. 2A-2C and 10 illustrate the handheld bioprinter 10 with the light curing module 18 removed.
  • the handheld bioprinter 10 can be coupled to (i.e., in fluid communication with) an array of external syringe pumps 26 for material extrusion.
  • each bioink cartridge 14 can be fluidly coupled to a syringe pump 60 via the flexible tube 24.
  • Each syringe pump 60 can be set to a specific pressure or specific flow rate that is optimized or selected for deposition of a specified type of bioink in the cartridge 14 that is coupled thereto.
  • the pressure or flow rate setting can be dependent on a viscosity of a bioink.
  • the pump array 26 is configured for hydraulic extrusion. Hydraulic extrusion can be particularly useful with low viscosity bioinks and can prevent material loss of low viscosity bioink through the printhead.
  • the pump array is configured for pneumatic extrusion.
  • FIGS. 11 A-l II and 12A-12I various printheads 12 that can be utilized with the handheld bioprinters disclosed herein, as well as corresponding configurations of bioink outputs from each printhead, are illustrated.
  • a printhead for use with the handheld bioprinters can be single channel, multi-compartment, multi-axial, or multi-core.
  • the printheads can be printed in photocurable resin using a Kudo3D Micro SLA 3D printer.
  • the differently configured printheads can each generate a specific configuration of bioink output or specific bioink material structure.
  • the multicompartment printheads can generate a side-by-side bioink deposition configuration, where each bioink is printed in substantially equal amounts and has an approximately equal distribution from a center to a perimeter of the deposit relative to other bioinks in the deposit.
  • the multi-axial printheads can generate a concentric bioink deposition configuration, wherein one or more bioink are printed within one or more other bioinks such that at least one bioink is enclosed inside of another bioink in a concentric manner.
  • the multi-core printheads can generate a radially arranged bioink deposition configuration, wherein two or more bioinks are enclosed inside of a primary bioink and are radially arranged (or other arrangement, such as aligned) inside of the primary bioink.
  • a channel of a printhead can be configured to produce a bioink deposit having a non-circular cross-section, for example a X- configuration cross-section, an oval cross-section, or a generally circular cross-section having an undulating (rather than smooth) edge.
  • any of the multi-compartment, multi-axial, or multi-core printhead examples can include a channel configured to generate a bioink deposit having a noncircular cross-section. It will be further appreciated that additional multi-compartment, multi-axial, and multi-core printhead configurations (such as combinations of the foregoing) are within the scope of the disclosure and can be designed for specific bioprinting applications.
  • single channel printheads can be utilized for drug delivery (FIGS. 14B and 14D), single-material biosensors (FIGS. 20C-20D), and/or handheld printing of single-material photocrosslinkable bioinks (FIGS. 16A-16I).
  • multi-compartment printhead can be utilized for multi-material handheld printing with quick-switching between materials (FIG.
  • coaxial printhead can be utilized for handheld bioprinting of chemically-crosslinkable bioinks (FIGS. 17A-17H), handheld bioprinting of materials that require sacrificial bioinks (i.e., pluronic) for structural integrity, and/or hollow, perfusable vascularized constructs (see FIG. 19).
  • multi-core printheads can be utilized for hollow, multi-channel perfusable vascularized constructs (FIG. 19G) and/or co-culture of different cell types in each core (for example, investigate tumor invasion and growth (FIGS. 18A-18F). Exemplary applications are discussed further below.
  • FIGS. 13A-20D illustrate various examples of handheld bioprinters and applications and methods of use for the handheld bioprinters disclosed herein.
  • FIG. 13A-13C include: (FIG. 13A i-xii) cross-sectional views and (FIG. 13A xiii-xv) photographic images of the different kinds of fibers produced with the different printhead configurations allowed by our platform; (FIG. 13B i-v) handheld bioprinted meshes on ex vivo pig skins with various wound shapes; and (FIG. 13C) photographic images of handheld bioprinter printing a multimaterial construct directly on an artificially made pig skin wound. As shown therein, the handheld bioprinter can perform in situ multi-material bioprinting on a curved or otherwise irregular surface.
  • FIGS. 14A-14D include: graphs showing cumulative release of FITC-loaded (FIG. 14A) and Rhodamine-loaded (FIG. 14B) PLGA microparticles embedded in PEGDA-laponite fibers with concentrations of 3, 6, and 9 mg/ml of microparticles in the fiber; (FIG. 14C) cumulative release of FITC-loaded thermoresponsive PNIPAAM microparticles embedded in PEGDA-laponite fibers, maintained at 20, 37, and 42 °C; and (FIG. 14D) a graph showing cumulative release of 3-material fiber with BSA, FITC-Dextran and Rhodamine-loaded PLGA microparticles, embedded in each side of the fiber. [0101] FIGS.
  • FIG. 15A-15C include: (FIG. 15A) schematic images of the cross-sectional view and the tip of the dual-material (SBS) printhead; and microscopy images of DAPI/Vimentin staining of (FIG. 15B) a monoculture of fibroblasts and (FIG. 15C) a co-culture of fibroblasts and HaCaT cells in each side of a GelMA-alginate fiber after (i) 1 day and (ii) 7 days of culture.
  • SBS dual-material
  • FIGS. 16A-16I include: (FIG. 16A) a photographic image of the device printing a 2-layer, fluorescent dye-loaded GelMA mesh, immediately photocrosslinked upon printing by the LED module; (FIG. 16B) a photographic image of a printed and crosslinked cell-loaded GelMA mesh; (FIG. 16C) side projection of the cell-loaded GelMA fiber; (FIG. 16D) Live/Dead microscopy images of a C2C12-loaded GelMA fiber after 1 day of culturing; DAPI/ Actin microscopy images of the printed cell-loaded GelMA meshes after 1 day (FIG. 16E) and 8 days (FIG. 16F) of culturing; (FIG. 16A) a photographic image of the device printing a 2-layer, fluorescent dye-loaded GelMA mesh, immediately photocrosslinked upon printing by the LED module; (FIG. 16B) a photographic image of a printed and crosslinked cell-loaded GelMA mesh; (FIG. 16C) side projection of the cell-loaded Gel
  • FIG. 16G Live/dead assay of CMs encapsulated in GelMA bioink after 7 days of incubation;
  • FIG. 16H DAPI/Actin/Troponin assay of CMs encapsulated in GelMA bioink after 5 days of incubation; and
  • FIG. 161) a graph showing relative fluorescence intensity of the cell- loaded GelMA meshes after 1 and 2 days of culturing.
  • FIGS. 17A-17H includes: (FIG. 17A) a schematic image of an exemplary printhead and bioink materials, and corresponding Live/dead assay of 3T3 Fibroblasts encapsulated in GelMA/Alginate bioinks using coaxial printhead after 1 day (FIG. 17 C) and 7 days (FIGS. 17D- 17E) of culture; and (FIG. 17B) a schematic image of an exemplary printhead and bioink materials, and corresponding live/dead assay after 7 days of culture (FIGS. 17F-17G), and DAPI/Actin/Troponin assay after 5 days of culture (FIG. 17H) of CMs encapsulated in GelMA/alginate bioinks using coaxial printhead.
  • FIGS. 18A-18C include: (FIG. 18A) schematic images of the cross-sectional view and the tip of the dual-core printhead; Live/dead fluorescent demonstrations of invaded SKOV-3 cancer cells in dual-core GelMA/Alginate fibers from day 1 to day 3 in (FIG. 18B) monoculture of SKOV- 3 cells, (FIG. 18C) Co-culture of SKOV-3/HNDF cells; (FIG. 18D) a photographic image of wetspun dual-core GelMA-aBlginate fibers, with each core loaded with either red or green fluorescent dye; (FIG.
  • FIGS. 19A-19G include: (FIG. 19A) schematic images of the cross-sectional view and the tip of the coaxial printhead; schematic images with accompanying photographic images of (FIG. 19B) a 2-dimensional hollow, perfusable fiber printed directly on an artificially created pig skin wound, (FIG. 19C) a 3-dimensional hollow, perfusable fiber, and (FIG. 19D) a hollow, perfusable fiber with varying diameter by varying printing speed; (FIGS. 19E-19F) photographic images of a hollow, perfusable fiber; and (FIG. 19G) a microscopy image a dual-channel hollow, perfusable fiber, wherein fibers were perfused with dye (red and/or green) for imaging.
  • FIGS. 19A-19G include: (FIG. 19A) schematic images of the cross-sectional view and the tip of the coaxial printhead; schematic images with accompanying photographic images of (FIG. 19B) a 2-dimensional hollow, perfusable fiber printed directly on
  • FIGS. 20A-20D include: (FIG. 20A) a photograph of handheld bioprinting of pH- responsive colorimetric biosensors on an artificially generated pig skin wound; (FIG. 20B) a photograph of the biosensors allowing for real-time detection of bacterial infection by changing color from clear (healthy skin) to blue (infected skin); and (FIGS. 20C-20D) photographs of conductive hydrogel fibers bioprinted on an artificially generated pig skin wound illuminating an LED when voltage is applied to the fibers.
  • the bioprinting system 100 can include a handheld bioprinter (extruder) 102 (for example, the bioprinter 10 described above) which can have a photocrosslinking module 114 (for example, the light module 18 described above) mounted thereon, an array of external syringe pumps 104 (for example, the syringe pump array 26 described above) in fluid communication with the handheld bioprinter 102 and configured for hydraulic or pneumatic bioink extrusion, a computerized controller 106, and a detection apparatus 110 that is configured to identify or measure one or more characteristics of bioink deposition in a deposition area or surface 112.
  • the bioprinter 102 can include one or more of the features of the exemplary handheld bioprinters described herein.
  • the computerized controller 106 can be in data communication with the bioprinter 102 (for example, the computerized controller can be in data communication with one or more actuators or controls 122 for dispensing bioink from the cartridges therein and/or a sensor 126 mounted on the bioprinter).
  • the computerized controller 106 can be in data communication with the photocrosslinking module 114 (for example, the computerized controller can be in data communication with one or more actuators or controls 128 for turning lights in a light array on and off).
  • the computerized controller 106 can be in data communication with the syringe pump array 104 (for example, the computerized controller can be in data communication with an actuator or control for each pump in the array).
  • the computerized controller can be in further data communication with the detection apparatus 110 and/or one or more external devices 108, such as a display and/or a user input device.
  • the computerized controller can be in data communication with additional or fewer components of the system and/or the system can include additional or fewer components.
  • the handheld bioprinter 102 can exclude an actuator and the computerized controller can be in data communication with the syringe pump array 104 for control of bioink deposition from the handheld bioprinter 102.
  • the system 100 can exclude a detection apparatus 110 or other detection mechanism (e.g., the sensor 126).
  • the computerized controller 106 can include one or more processors 116, a data communication interface 118, and (non-transitory) memory 120 having one or more programs stored thereon that are configured to be executed by the one or more processors.
  • the memory 120 can store application specific programming (such as for the applications illustrated in FIG. 13-20 or other applications) for deposition of bioinks in specified protocols, recipes, spatial and/or temporal patterns, etc.
  • the computerized controller 106 can control the flow of bioink via controlling the syringe pump array 104 and/or the actuators 122 on the handpiece 102.
  • the computerized controller 106 can further control photocrosslinking via operation of the photocrosslinking module 114.
  • a sensor 124 of the detection apparatus 110 can detect one or more characteristics of the bioink deposition in the deposition area 112 and the detection apparatus 110 can send signals or data to the controller 106. Additionally or alternatively, in some examples, an onboard sensor 126 disposed within or attached to the handpiece 102 can detect one or more characteristics of the bioink deposition in the deposition area 112 and/or one or more characteristics of the bioink as it is flowing from the printhead onto the deposition area 112.
  • the programming can be configured to determine, based on the signals/data from the detection apparatus 110 and/or the signals/data from the onboard sensor 126, whether the deposition meets specified parameters (e.g., position, size, flow rate, etc.), or if flow from one or more of the bioink cartridges should be adjusted.
  • the computerized controller can adjust the one or more controls on the syringe pump array 104 and/or the actuators 122 on the handpiece 102.
  • the senor 124 is configured to detect one or more of flow of bioink, and/or a position, a configuration, and/or a structure of bioink dispensed from a printhead of the handpiece 102, and the detection apparatus 110 is in signal communication with the computerized controller 106 and configured to provide feedback data thereto.
  • the sensor 126 is configured to detect one or more of flow of bioink and/or a position, a configuration, and/or a structure of bioink dispensed from the multi-channel printhead of the handpiece 102, and the sensor 126 and/or a microcontroller within the handpiece 102 is in signal communication with the computerized controller 106 and configured to provide feedback data thereto.
  • the computerized controller is configured to, based at least on the feedback data indicating insufficient dispensement of bioink for one or more bioink cartridges coupled to the printhead, increase the respective pressure setting or flow rate for the syringe pump in fluid communication with the identified cartridge and/or adjust the corresponding cartridge actuator.
  • the computerized controller is further configured to, based at least on the feedback data indicating over dispensement of bioink for one or more bioink cartridges, decrease the respective pressure setting or flow rate for the syringe pump in fluid communication with the identified cartridge and/or adjust the corresponding cartridge actuator.
  • the bioprinting system 200 can include a bioprinter (extruder) 202 (for example, the bioprinter 10 described above) which can have a photocrosslinking module 214 (for example, the light module 18 described above) mounted thereon, an array of external syringe pumps 204 (for example, the syringe pump array 26 described above) in fluid communication with the bioprinter 202 and configured for hydraulic or pneumatic bioink extrusion, a computerized controller 206, and a detection apparatus 210 that is configured to identify or measure one or more characteristics of bioink deposition in a deposition area or surface 212.
  • the bioprinter 202 can include one or more of the features of the exemplary handheld bioprinters described herein.
  • the computerized controller 206 can be in data communication with the bioprinter 202 (for example, the computerized controller can be in data communication with one or more actuators or controls 222 for dispensing bioink from the cartridges therein and/or a sensor 226 mounted on the bioprinter).
  • the computerized controller 206 can be in data communication with the photocrosslinking module 214 (for example, the computerized controller can be in data communication with one or more actuators or controls of the photocrosslinking module 214 for turning lights in a light array on and off).
  • the computerized controller 206 can be in data communication with the syringe pump array 204 (for example, the computerized controller can be in data communication with an actuator or control for each pump in the array). In some examples, the computerized controller can be in further data communication with the detection apparatus 210 and/or one or more external devices 208, such as a display and/or a user input device.
  • the system 200 can further include a single-axis or a multiaxis positioning systems 228 (such as, for example, a robotic arm or a high performance positioning system) coupled to the bioprinter 202 and/or a single-axis or a multi-axis positioning systems 232 (such as, for example, a robotic arm or a high performance positioning system) coupled to the deposition surface 212.
  • a single-axis or a multiaxis positioning systems 228 (such as, for example, a robotic arm or a high performance positioning system) coupled to the deposition surface 212.
  • Positioning and movement of the positioning system 228 (as well as the bioprinter 202 coupled thereto) can be controlled along one or a plurality of axes by, for example, one or a plurality of motors and/or solenoids 230.
  • Positioning and movement of each the positioning system arm 228 can be controlled along one or a plurality of axes by, for example, one or a plurality of motors and/or solenoids 234.
  • Each of the motors/solenoids 230 and 234 can be in data communication with the computerized controller 206.
  • the system 200 can include two or more bioprinters 202 each comprising a printhead and configured for coupling to a plurality of cartridges. Further, each of the bioprinters 202 can be coupled to a single-axis or a multi-axis positioning systems 228 and corresponding motors or solenoids 230 and can be independently positionable relative to the other bioprinters of the system.
  • the computerized controller can be in data communication with additional or fewer components of the system and/or the system can include additional or fewer components.
  • the bioprinter 202 can exclude an actuator and the computerized controller can be in data communication with the syringe pump array 204 for control of bioink deposition from the bioprinter 202.
  • the system 200 can exclude a detection apparatus 210 or other detection mechanism (e.g., the sensor 226).
  • the system 200 can include just one of the robotic arms 228 and 223 and corresponding motors 230 and 234.
  • the computerized controller 206 can include one or more processors 216, a data communication interface 218, and (non-transitory) memory 220 having one or more programs stored thereon that are configured to be executed by the one or more processors.
  • the memory 220 can store application specific programming (such as for the applications illustrated in FIG. 13A-20D or other applications) for deposition of bioinks in specified protocols, recipes, spatial and/or temporal patterns, etc.
  • the computerized controller 206 can control the flow of bioink via controlling the syringe pump array 204 and/or the actuators 222 on the bioprinter 202.
  • the computerized controller 206 can further control photocrosslinking via operation of the photocrosslinking module 214.
  • the computerized controller 206 can further control positioning and movement of bioprinter 202 via controlling the motor 230 and the robotic arm 228. In some examples, the computerized controller 206 can further control positioning and movement of the deposition surface 212 via controlling the motor 234 and the robotic arm 232.
  • a sensor 224 of the detection apparatus 210 can detect one or more characteristics of the bioink deposition in the deposition area 212 and the detection apparatus 210 can send signals or data to the controller 206. Additionally or alternatively, in some examples, an onboard sensor 226 disposed within or attached to the bioprinter 202 can detect one or more characteristics of the bioink deposition in the deposition surface 212 and/or one or more characteristics of the bioink as it is flowing from the printhead onto the deposition surface 212.
  • the programming can be configured to determine, based on the signals/data from the detection apparatus 210 and/or the signals/data from the onboard sensor 226, whether the deposition meets specified parameters (e.g., position, size, flow rate, etc.), or if flow from one or more of the bioink cartridges and/or positioning and movement of the bioprinter 202 and/or the deposition surface 212 should be adjusted.
  • the computerized controller can adjust the one or more controls on the syringe pump array 204 and/or the actuators 222 on the bioprinter 202.
  • the computerized controller can adjust the motors 230 and/or 234.
  • the senor 224 is configured to detect one or more of flow of bioink, and/or a position, a configuration, and/or a structure of bioink dispensed from a printhead of the bioprinter 202, and the detection apparatus 210 is in signal communication with the computerized controller 206 and configured to provide feedback data thereto.
  • the sensor 226 is configured to detect one or more of flow of bioink and/or a position, a configuration, and/or a structure of bioink dispensed from the multi-channel printhead of the bioprinter 202, and the sensor 226 and/or a microcontroller within the bioprinter 202 is in signal communication with the computerized controller 206 and configured to provide feedback data thereto.
  • the computerized controller can be configured to, based at least on the feedback data indicating insufficient dispensement of bioink for one or more bioink cartridges coupled to the printhead, increase the respective pressure setting or flow rate for the syringe pump in fluid communication with the identified cartridge and/or adjust the corresponding cartridge actuator.
  • the computerized controller is further configured to, based at least on the feedback data indicating over dispensement of bioink for one or more bioink cartridges, decrease the respective pressure setting or flow rate for the syringe pump in fluid communication with the identified cartridge and/or adjust the corresponding cartridge actuator.
  • the computerized controller can be configured to, based at least on the feedback data indicating undesired location, structure or configuration of bioink, adjust a position or a speed of movement of one or more of the bioprinter 202 and/or the deposition surface 212.
  • the bioprinter(s) 202 in the system 200 can include a passive temperature control module (such as the temperature control modules discussed above with reference to FIGS. 3 and 4).
  • the system 200 can include other or alternate temperature control mechanisms, such as, for example, a closed environment control system to configured to control a temperature and/or humidity within an enclosure housing the bioprinters and the deposition surface or a coolant circuit associated with each of the bioprinters in the system configured to circulate fluid of a specified temperature around the bioink cartridges loaded in a respective bioprinter.
  • systems 100 and 200 can be adapted for other uses and applications and/or can include additional components, such as the uses, applications, and components discussed below with reference to FIGS. 24-29C.
  • bioprinter 10 can be adapted for other uses and applications and/or can include additional components, such as the uses, applications, and components discussed below with reference to FIGS. 24-29C.
  • an exemplary hydraulic cartridge 300 which can include one or more features of the cartridges 14 and/or can be used in the bioprinters disclosed herein in place of the cartridges 14, is shown and described.
  • the hydraulic cartridge 300 can serve as a primary extrusion mechanism, responsible for the precise deposition of bio-inks and other biological materials, setting them apart from more known methods such as direct syringe pump extrusion or pneumatic extrusion.
  • the hydraulic cartridge 300 includes a tube 302 (also referred to as a “barrel”, a “tubular main body”, and a “reservoir”), a piston 304 in the tube 302, an adapter 306 (also referred to as a “sealing mechanism”), a tube 308 (for example, a polytetrafluoroethylene (PTFE) tube) for connecting the cartridge 300 to a syringe pump (such as one of the syringe pumps disclosed herein), and a connector 310.
  • a tube 302 also referred to as a “barrel”, a “tubular main body”, and a “reservoir”
  • an adapter 306 also referred to as a “sealing mechanism”
  • a tube 308 for example, a polytetrafluoroethylene (PTFE) tube
  • the connector 310 can be a Luer lock connector.
  • the connector 310 can be a male Luer connector that is configured to be received in a female Luer connector on a printhead, such as the printheads 12 discussed above, and/or the printhead 412, 512 shown in FIGS. 25A-25C.
  • the male Luer connector can include a threaded skirt (which can be a rotatable coupling) for threaded engagement with the female Luer connectors on a printhead.
  • the Luer connectors on the cartridge and the printhead can be male and female tapered connectors that can be press fit together.
  • the hydraulic cartridge 300 operates based on hydraulic force.
  • a driving fluid for example, water or oil, such as mineral oil, water-glycol hydraulic fluids or biodegradable hydraulic fluids
  • a driving fluid for example, water or oil, such as mineral oil, water-glycol hydraulic fluids or biodegradable hydraulic fluids
  • This method can offer improved control over the speed and volume of extrusion and/or improved accuracy and precision of bioprinting.
  • one advantage of the hydraulic cartridge 300 over other extrusion methods is the level of precision and control they offer.
  • the hydraulic cartridge 300 can offer greater flexibility and ease of use.
  • direct syringe pump systems are typically bulky and not easily adaptable to a handheld device.
  • a system including hydraulic cartridges 300 can be more compact and designed for integration with a handheld bioprinter (such as, the bioprinter 10 and/or other bioprinters disclosed herein), making the technology more accessible and user-friendly over known systems.
  • FIGS. 25A-25C show an exemplary multi-material mixing printhead 412 (which can have one or more features of the printheads 12 discussed above, and can be utilized with the bioprinters disclosed herein, such as the bioprinter 10 and/or the bioprinters 410, 510 in the systems 400, 500 shown in FIGS. 26-29C).
  • the mixing printhead 412 can be configured to combine different bio-inks (or other flowable materials) in controlled proportions and/or sequences, thereby enabling, for example, fabrication of complex, multi-layered tissues and/or other structures.
  • the printhead 412 can include a base portion 430 comprising an inflow interface configured to enable coupling of bioink cartridges 414 thereto (for example, configured to enable coupling of two, three, or more bioink cartridges thereto), a tip portion 432 having a distal opening 434 configured for output of bioink, and bioink feed channels 428a that extend through the base portion 430.
  • a bioink mixing channel 428b can extend through the tip portion 432 to enable flow of bioink from the bioink cartridges through the base and tip portions to the opening 434.
  • the cartridges 414 can be and/or can have one or more of the features of the cartridges 14, 300 discussed above.
  • cartridges such as the cartridges 300 can be coupled to the base portion 430 by securing the male Luer lock connectors (such as, the exemplary connector 310) of the cartridges to the female Luer lock connectors 431 incorporated into the base portion 430.
  • the feed channels 428a can be separate channels each configured for communication with one of the cartridges 414 to enable flow of bioink (or another flowable material) from the respective cartridge 414 to the bioink mixing channel 428b.
  • the bioink mixing channel 428b can be a common channel configured for mixing the bioinks (or other flowable materials) as they flow through the mixing channel 428b toward the distal opening 434.
  • the bioink mixing channel 428b can include a structure for causing turbulence and/or a non-linear flow pathway for materials moving therethrough, such as a corkscrew member 470 (which can also be referred to as a “mixer”, a “turbulator”, and a “helical member”) including a plurality of spaced openings 472, which can be, for example, located at each half-turn or each turn of the corkscrew member 470.
  • the mixing channel 428b can include other structures or configurations for causing turbulence and/or a non-linear flow pathway for materials moving therethrough.
  • the mixing printhead 412 can be coupled to a bioprinter 410 (which can have one or more of the features of the bioprinter 10 discussed above or other features), which can be in communication with a hydraulic pump array 426 (which can have one or more of the features of the hydraulic pump array 26 discussed above or other features) via tubes 424 (which can have one or more of the features of the tubes 24 discussed above or other features).
  • a bioprinter 410 which can have one or more of the features of the bioprinter 10 discussed above or other features
  • a hydraulic pump array 426 which can have one or more of the features of the hydraulic pump array 26 discussed above or other features
  • tubes 424 which can have one or more of the features of the tubes 24 discussed above or other features.
  • One benefit of the mixing printhead 412 is its capacity to mix bio-inks just prior to deposition. This can enable the bio-inks to remain separate until the point of printing/deposition, thereby preventing premature crosslinking or interactions that could impact the quality or functionality of the final bioprinted structure. This feature may also enable bioinks with different properties (like viscosity or gelation time) to be optimally printed according to the material properteis.
  • the mixing printhead 412 can enable diverse applications of a handheld bioprinter, for example, to applications beyond biomedical applications, such as arts and cosmetics for precision color mixing.
  • the printhead can facilitate automatic, precise color mixing, relieving artists from the often time-consuming manual process, and can enable exact color matches and improve consistent reproducibility.
  • the ability to create a wide array of shades and gradients opens up new dimensions for artistic creativity.
  • the precise deposition also facilitates the generation of custom textures and intricate details, augmenting the realm of artistic possibilities.
  • the mixing printhead can provide a mechansim to create personalized color cosmetics, which can accurately match and replicate skin tones for products such as foundations and concealers.
  • the mixing printhead can enable formulation of custom shades of various cosmetic products like lipsticks, eyeshadows, and blushes, enhancing customization options.
  • the mixing printhead 412 can enable additional benefits in automating color switching, particularly advantageous for applications such as airbrushing in the arts and cosmetic applications. For example, airbrushing often demands a change of colors to create desired artistic effects. Traditionally, this involves a manual process of cleaning the airbrush tool each time a color switch is necessary. However, with the multi-material mixing printhead 412, artists can switch between different colors without the need for tedious and time-consuming manual cleaning.
  • the mixing printhead 412 is designed to contain multiple color cartridges, each loaded with a different color. By automatically switching between cartridges, or even mixing multiple ones at once, artists can achieve fast, efficient color changes. Airbrushing techniques may also be used for applying makeup in cosmetic applications.
  • the multi-material mixing printhead 412 can significantly enhance this process by allowing for a swift and precise change of colors. Makeup artists can load different shades of foundations, blushes, or eye shadows into separate cartridges within the printhead. With the ability to switch between these colors instantaneously, the makeup application process can be more efficient and precise.
  • FIGS. 27A-29C a robotic bioprinter system 500 and an assembly 501 for adapting a 3D printer 502 to the robotic bioprinter system 500 is shown and described.
  • the assembly 501 can be a conversion kit designed to adapt a 3D printer 502 into a robotic bioprinter system 500.
  • the 3D printer 502 includes a platform 503 and a robotic positioning assembly 504.
  • the assembly 501 can include one or more of the printheads, light control modules, temperature control modules, and/or the hydraulic extrusion pump arrays disclosed herein. In one example illustrated in FIGS.
  • the assembly 501 includes a hydraulic pump array 526, a carriage 542, a bioprinter 510 (including cartridges 514 and a printhead 512, which can be the cartridges 14, 300 and the printheads 12, 412 and/or can have one or more of the features of the cartridges 14, 300 and the printhead 12, 412 discussed above), and a user interface 505.
  • the bioprinting system 500 can be used for automated printing of a 3D construct 538’ from bioinks and/or other flowable materials 538 dispensed from the cartridges 514.
  • the platform 503 can be movable beneath the printhead 512 along guide members 507 oriented in the direction of the y-axis in FIG. 27A.
  • the assembly 501 can be designed with simplicity and usability to enable users to upgrade an existing 3D printer.
  • the assembly 501 can include a non-transitory storage or memory having one or more computer-readable instructions stored thereon, which are configured to control operation of the 3D bioprinter system.
  • the assembly 501 can include an open-source interface (firmware) that is compatible with many 3D printer models.
  • the firmware can enable users to control the bioprinting functionalities of a converted 3D printer, such as controlling hydraulic syringe pumps and/or an LED module.
  • firmware for the assembly 501 can be based on Klipper firmware, which can support benefits of the system, such as being high-precision, highly extensible and configurable, and/or being configured for multi-controller support (needed for multiple syringe pumps).
  • the assembly 501 can include a custom profile for compatibility with the most common open-source slicer (such as, for example, Ultimaker Cura®).
  • the assembly 501 can include custom post-processing software to automatically process the sliced model for different types of printing, such as, for example, directly on bed and/or FRESH printing.
  • the assembly 501 can include a graphical user interface (GUI) 505, which can be, for example, a mountable touchscreen GUI (for example, a Rasberry Pi and Touchscreen), which can be Wi-Fi enabled for wireless control via a separate user device, such as a mobile device or a user’s computer.
  • GUI graphical user interface
  • the hydraulic syringe pump array 526 can have one or more of the features of the pump array 26 discussed above and/or other features.
  • each pump 528 in the array 526 can include an actuator 530, such as, for example, a stepper motor-driven linear stage actuator, which can provide a level of precision with specified accuracy and reproducibility for applications such as bioprinting, color mixing in arts, and/or cosmetics.
  • the syringe pumps 528 in the array 526 can be compact and have a small form factor.
  • each of the syringe pumps 528 can include the actuator 530, a syringe 533 having a plunger 541 (also shown in FIG. 27B) disposed therein, one or more covers or housings 534, a pusher block 535, position sensors 536, 537, and a support structure or mount 540 configured to support and/or have the syringe 533 mounted thereto.
  • the actuator 530 can be a stepper motor-driven linear stage actuator, including a stepper motor 531 and a linear stage actuator 532.
  • the linear stage actuator 532, the stepper motor 531, the support structure 540, and the syringe 533 remain stationary, and the pusher block 535 is moveable relative thereto and can apply a downward force on a plunger of the syringe 533.
  • the linear actuator 532 can include an externally threaded member or shaft 539 that can be threadedly engaged with internal threads of the pusher block 535, and rotation of the externally threaded member (driven by the stepper motor 531) can drive axial movement of the pusher block 535. Downward axial movement of the pusher block 535 can bring the pusher block into contact with a top (actuation) surface of the plunger and can drive the plunger downward and further into the syringe 533.
  • FIG. 28B shows the pump 528 with the covers 534 removed, and shows locations of the positional sensors 536, 537.
  • the positional sensors 536, 537 can be an encoder, a potentiometer, a linear variable differential transformer, and/or other types of position and/or motions sensors.
  • the positional sensors 536, 537 can include or can be coupled to switches on the pusher block 535 and/or other parts of the assembly.
  • the sensor 537 can be in electrical communication with a switch on a lower surface of the pusher block 535 can receive a signal when the pusher block contacts the plunger of the syringe and depresses the switch.
  • the positional sensors 536, 537 can include and/or can be substituted with end stop switches including, for example, a mechanical lever or button.
  • the positional sensor 536 can serve to identify the "zero" or "top” position of the pusher block 535 and can stop further axial movement (e.g., upward movement) of the pusher block upon activation of the positional sensor 536.
  • the positional sensor 537 is mounted on the pusher block 535.
  • a face of the positional sensor 537 is located on a bottom surface of the pusher block 535 and can contact the top surface of the plunger. In some examples, the positional sensor 537 can detect a location of the plunger within the syringe 533. In some examples, the positional sensor 537 can reduce or eliminate the need for manual calibrations of the syringe pump 528.
  • the positional sensor 537 can serve to identify a “bottom” position of the pusher block 535 (for example, a position where the plunger is inserted into the syringe at a maximum desired insertion) and can stop further axial movement (e.g., downward movement) of the pusher based on signals received from (e.g., closing of an electrical circuit) of the positional sensor 536.
  • the carriage 542 can be configured to be coupled to the robotic positioning assembly 504 of the 3D printer 502 (FIGS. 29A-29C).
  • the carriage 542 and/or the platform 503 can include end stop switches for limiting movement of the carriage and/or the platform.
  • the carriage 542 and/or the platform 503 (or other components of the printer, such as the frame) can include end stop switches comprising a mechanical lever or button. When the carriage 542 or the platform 503 reaches one of these switches, it can press against the lever or button, thereby causing it to actuate and/or close an electrical circuit.
  • the switch When the switch is triggered or activated, it can send a signal to a computerized controller and/or motherboard of the 3D printer to indicate that the printer (e.g., the carriage 542 and/or the platform 503) has reached its limit along one or more of the axes (i.e., x, y, and/or z).
  • the controller receives the signal, it will identify or generate a “stop” command for the respective axis.
  • the controller can then instruct the printer’s motors (in the robotic positioning assembly 504) to halt or reverse direction, preventing the carriage 542 and/or the platform 503 from crashing into each other, the printer frame, and/or other components, which could damage the printhead, the printer, and/or the printed construct.
  • the carriage 542 can include a base 544 comprising channels 546 configured receive portions (for example, X-axis rails or guide members 543) of the robotic positioning assembly 504 for mounting the carriage 542 to the robotic positioning assembly 504.
  • the carriage 542 can also include a plate member 548 that is attached to the base 544 and a cartridge holder 550 (which can also be referred to as an “adapter”) that can be releasably coupled to the plate member 548.
  • the plate member 548 and the cartridge holder 550 can be configured for magnetic coupling, and can include complementary magnets 552, 554 in the plate member 548 and the cartridge holder 550.
  • the cartridge holder 550 includes recesses 556 configured to receive cartridges 514.
  • the cartridge holder 550 can be made of a deformable material or elastic material such that the cartridges 514 can be retained within the recesses 556 via a compression fit therein for coupling the bioprinter 510 to the carriage 542.
  • the carriage 542 and the bioprinter 510 can move (e.g., translate) along guide members 543 of the positioning assembly 504.
  • the carriage 542 can thus position the bioprinter 510 (including the printhead 512) along the x-axis (FIG. 29A) with respect to the platform 503 and workpiece(s) on the platform.
  • the carriage 542 and bioprinter 510 can be moved along the z-axis by modules 545 configured to translate up and down along guide members 547.
  • the magnetic attachment between the plate member 548 and the cartridge holder 550 can allow for easy and/or rapid changes between different cartridges and/or printheads, catering to needs of a variety of applications.
  • the magnetic mount can act as a safety mechanism.
  • the cartridge holder 550 can detach without causing damage to the printer, cartridges, or printhead, which can reduce the risk of hardware damage during operation, enhancing user experience and the overall reliability of the system.
  • the plate member 548 and the cartridge holder 550 can be configured for other types of releasable coupling (for example, releasable snap-fit coupling).
  • the assembly the assembly 501 can be configured for adapting an existing 3D printer to the bioprinter system 500.
  • a heatable head for filament heating and extrusion can be uncoupled or removed from the robotic positioning assembly 504 and can be replaced with the carriage 542 (for example, by coupling the carriage 542 to the X- axis rails).
  • the hydraulic syringe pump array can be coupled to a top portion of a frame of the 3D printer.
  • the user interface 505 can be coupled to a side portion of the frame of the 3D printer, and can be brought into data communication with the 3D printer, for example, via one or more wires or wireless data communication (e.g., WiFi, WLAN, etc.).
  • wires or wireless data communication e.g., WiFi, WLAN, etc.
  • the bioprinter system 500 can be operated by (i) selecting a printhead based on a desired application, (ii) coupling one or more bioink cartridges to the printhead (for form an assembled bioprinter), (iii) optionally enclosing the bioink cartridges in a housing and/or a temperature control module, (iv) optionally attaching a light curing module to the bioprinter, (v) coupling the bioprinter to the carriage, (vi) selecting pressure settings and/or flowrate for the hydraulic syringe pumps based one or more criteria (such as, for example, a type of bioink and/or a print application), and (vii) selecting a stored program for a selected printing application (for example, computer-readable instructions that control x-axis and/or z-axis movements of the robotic positioning assembly and/or y-axis movements of the bioprinter platform), entering data for a selected printing application, and/or entering a command to start printing, which can each be selected
  • the robotic bioprinter system 600 includes a bioprinter 610 (which can have one or more of the features of the bioprinters, such as, for example, the bioprinter 10 and/or other bioprinters disclosed herein) and a multi-axis robotic manipulator arm 602, which can be, for example, a six- axis robotic arm (for example, a Dorna 2 robotic arm).
  • the robotic manipulator arm 602 is schematically illustrated in FIG. 30. Although not shown, the robotic manipulator arm 602 can be coupled to a gantry or other support frame.
  • the robotic manipulator arm 602 can include a plurality of links and can be movable in multiple degrees of freedom (e.g., six degrees of freedom).
  • the bioprinter 610 is an end effector mounted on a distal end of the robotic manipulator arm 602.
  • the end effector positions and velocities can be controlled digitally via a computerized controller 601 in data communication with the robotic manipulator arm 602.
  • each motion of each of the plurality of links e.g., each degree of freedom (DOF) of the multi-axis robotic manipulator arm 602 can be controlled using a separate position control system.
  • the motions can coordinated by the computerized controller 601.
  • the computer controller can also provide an interface (GUI) between the robotic manipulator arm 602 and an operator for receiving user input from and providing information to the operator.
  • GUI interface
  • the robotic bioprinter system 600 can be utilized in surgical applications.
  • the robotic bioprinter system 600 can be a surgical tool for minimally invasive, image-guided, robotic-assisted in vivo biomaterial delivery, which can, for example, be used to deliver specifically formulated (e.g., ‘coded’) hydrogel fibers into a targeted treatment site, such as a site of injury or disease or other condition on or within a patient.
  • a multichannel printhead (such as, for example, one or more of the printheads shown in FIGS. 11 A-l II, 12A-12I, and 25A-25C) can be utilized in the bioprinter 610 and can, for example, deliver one or more bioinks on a microscale that can be used for in situ multi-material bioprinting.
  • a bioink and/or a combination of bioinks can be optimized for a particular application and/or for use with a specific printhead.
  • bioink extrudability, form integrity, and filament characterization can be used to determine printability of the bioink(s) for a specific application (such as, for example, repair of a specific anatomical structure and/or treatment of a specific tissue type).
  • bioink concentration and molecular weight, infill density, and/or printing pattern can be selected to achieve modulus in the range of 1-10 kPa, which can cover the range of stiffness observed in various cardiac tissues.
  • the system 600 can integrate the bioprinter 610 into a surgical assembly by connecting the bioprinter 610 (such as, fluidly coupling the printhead thereof) to a catheter 616 to enable delivery of the injected bioink constructs (e.g., fibers) directly into and/or onto an internal anatomical structure 606 (such as, for example, a heart, as illustrated in FIG. 30, or an artery, a blood vessel, a kidney, a lung, etc.) under guidance of an optical imaging system.
  • an internal anatomical structure 606 such as, for example, a heart, as illustrated in FIG. 30, or an artery, a blood vessel, a kidney, a lung, etc.
  • the system 600 can include fiber-based optical coherence tomography (OCT) imaging guidance via an OCT assembly 608 in communication with the controller 601.
  • OCT optical coherence tomography
  • the OCT assembly 608 can be an intravascular imaging system that uses near-infrared light to provide high-definition, cross-sectional and/or three-dimensional images of the anatomical (for example, vascular) microstructure.
  • the OCT assembly 608 can provide closed-loop feedback to enable real-time positioning instruction and/or positioning correction of the end effector (the bioprinter 610) on a dynamic anatomical printing surface (such as, for example, a beating heart or other anatomical structure subject to active or passive movement).
  • the OCT assembly 608 can be configured for (i) submillimeter-level precision to minimize errors that may cause a collision of the printhead into the tissue and/or the bioink construct to prevent injury to the tissue and/or damage to the bioink construct, (ii) a short sensing range (for example, approximately 10 mm) in order to accommodate a restricted workspace, and/or (iii) a high heart wall positioning reconstruction rate (for example, >1000 Hz), which may be an order of magnitude faster than the R-wave peak (for example, approximately 35 ms) and three orders of magnitude faster than the adult human heart beating rate at rest (for example, approximately 60 beats/min).
  • the heart wall positioning feedback can be used as input for the controller 601 of the robotic manipulator arm 602 for adaptive 3D bioprinting.
  • the system 600 can be a closed-loop system including: (i) miniaturized fiber-based OCT probes 612, (ii) an OCT engine 614, and (iii) the robotic arm 602.
  • the OCT fiber probes 612 can be attached to and/or extend through a catheter 616.
  • the probes 612 can be comprised of three optical fiber components (each 0.125 mm in diameter) spliced together, such as, for example, (i) a single mode fiber (SMF), (ii) a coreless fiber (CF), and (iii) a gradient index (GRIN) fiber.
  • the fiber optic probes 612 can enable distal sensing with, for example, micrometric resolution.
  • an exit port of the printhead in the bioprinter 610 can connect to and/or can be in fluid communication with a microinjection catheter 618, which can extend through the (outer) catheter 616.
  • Bioink flow from the bioprinter 610 can be controlled by connecting syringe pumps in a syringe pump array 626 (which is schematically illustrated in FIG. 30 but can be similar to the syringe pump arrays 26, 526 disclosed herein or have other features) in communication with and/or attached to a computer-programmable valve system 627.
  • a bioink quantity, type, and/or concentration in a bioink construct can be controlled via the valve system 627.
  • photosensitive bioinks can be cross-linked with light module or fiber (not shown) within the catheter 616 upon exit.
  • the system 600 can be utilized to generate a desired bioink construct architecture.
  • a bioink fiber can be extruded in which the material properties are controlled along its length.
  • the system 600 can be utilized to generate meter-long woven, braided and/or knitted multifunctional microfibers with controllable biophysical-chemical features.
  • the system 600 can be utilized to generate Janus fibers in which one side of the fiber (e.g., one part of the fiber cross section) is different from the other side, enabling co-delivery of various cells and/or growth factors.
  • each generated microfiber can vary in size, shape, and/or physical and chemical structure, depending on its function within the tissue and/or its roll in tissue repair.
  • the fabricated fibers for example, fibers having a diameter in a range of 10 to 30 microns and a length in a range of 0.5 to 3 cm
  • the fabricated fibers can self-assemble into structures (up to, for example, 30 mm in size).
  • In vitro tests can be performed prior to treatment to quantify selfassembly of spatially patterned cross-linked fibers and establish ex vivo models (for example, a beating heart model (using porcine heart)) to optimize the printing conditions and refine and/or train the optical tracking system (e.g., the OCT assembly 608).
  • the system 600 can be used in combination with or can include other surgical tools and/or assemblies.
  • the system 600 can be used in combination with or can include a trocar 620 for enabling access to the treatment site through a tissue corridor and/or a stabilizer device 622, such as, for example, a device for stabilizing a position of the heart.
  • the bioprinter and bioprinting systems disclosed herein can have other applications.
  • the bioprinting systems disclosed herein can be used for: automated paint mixing (for example, the bioprinters disclosed herein, with a mixing printhead, could be used to automatically mix paints, enabling artists to quickly create and reproduce exact colors with high precision, and/or can be used to allow artists to use their phone camera as a “color picker” to be replicated with the mixer); airbrushing (for example, the bioprinters disclosed herein can be used for rapid and seamless color-switching capabilities could make it a powerful tool for airbrushing, and/or artists could create multi-color designs more efficiently, since currently switching colors requires emptying and cleaning the airbrush and paint reservoir); custom paint textures (for example, by controlling and fine-tuning the properties of the inks, artists could create custom textures in their paintings, adding depth and complexity to their work utilizing
  • the bioprinting systems disclosed herein can be used for: personalized cosmetics (for example, the bioprinters disclosed herein could be used to print personalized skincare products directly onto the skin, such as products tailored to an individual's skin type, specific needs, or preferences); personalized face masks (for example, a user could input information about their skin type and concerns (dryness, acne, redness, etc.) into a computerized application connected to the bioprinters disclosed herein, and the computerized application could then determine the optimal mix of bioinks (e.g., those containing moisturizing hyaluronic acid for dry skin, anti-inflammatory ingredients for acne-prone skin, or calming agents for irritated skin) and the device could then print this custom mask directly onto the user's face, with different areas of the face receiving treatments as needed); color cosmetics (for example, utilizing the bioprinters disclosed herein with a mixing printhead, a foundation that matches a user's unique skin tone, and/or other printhead types could be used to create
  • personalized cosmetics for example, the bioprinters
  • the bioprinting systems disclosed herein can be used for: customized nutrition (for example, the bioprinters disclosed herein can be used to create food items with customized nutritional content based on the specific needs of an individual, such as, for example, meals designed to have specific amounts of proteins, carbs, vitamins, etc., to match a person's dietary requirements; food texture modification (for example, the bioprinters disclosed herein can be used to create foods with modified textures, which may be particularly beneficial in creating appetizing and nutritious foods for people who have difficulty swallowing or chewing); alternative proteins (for example, the bioprinters disclosed herein can be used to create meat substitutes from plant-based proteins or lab-grown animal cells, assisting in meeting the growing demand for environmentally friendly and ethical meat alternatives; and/or personalized shapes and sizes (for example, the bioprinters disclosed herein can allow consumers or businesses to create food in personalized shapes and sizes, which could be particularly appealing in the confectionery or pastry sectors).
  • customized nutrition for example, the bioprinters disclosed herein can be used to create food items with customized nutritional content based on the specific needs
  • the printhead 412 can be configured to receive five bioink cartridges as shown, or can be configured to receive any other number of bioink cartridges in any of the arrangements described herein.
  • the 3D bioprinter 500 can be configured to include one or more of the printhead examples, one or more of the bioink cartridge examples, and/or one or more of the hydraulic syringe pump examples described herein.
  • Described here are a modular handheld bioprinter that can deposit a variety of bioinks in situ with precise control over their physical and chemical properties is disclosed, as well as other bioprinter systems and assemblies.
  • the ergonomic design of the handheld bioprinter facilitate the shape-controlled biofabrication of multi-component fibers with different cross-sectional shapes and material compositions.
  • the capabilities of the produced fibers in local delivery of therapeutic agents was demonstrated by incorporating drug-loaded microcarriers, extending the application of the printed fibers to on-demand, temporal, and dosage-control drug delivery platforms.
  • the versatility of this platform to produce biosensors and wearable electronics was demonstrated via incorporating conductive materials and integrating pH responsive dyes.
  • the efficacy of this handheld printer to generate cell-laden fibers with high cell viability for site-specific cell delivery was shown by producing single component and multicomponent cell-laden fibers.
  • the multi-component fibers were able to model the invasion of cancer cells into the adjacent tissue.
  • bioprinted constructs can be controlled through rational design of bioinks and selection of the most suitable printing method(s) to mimic the native tissue function. Indeed, recapitulating the multiscale complexities of tissues requires advanced technologies that involve high resolution printing of multiple bioinks, creating sophisticated multi-luminal structures.
  • bioprinters capable of depositing multiple materials with control over their spatiotemporal physicochemical properties.
  • This modular bioprinter is equipped with a mountable light-emitting compartment that facilitates printing photo-crosslinkable bioinks.
  • the bioprinter can include a passive temperature control module that maintains the temperature of cartridges for extended periods, allowing for long-term printing of viscous bioinks.
  • Multi-material printing is achieved by means of multi-channel printheads with complex fluidic circuitry that enable creating complex flows of several bioinks in microscale.
  • the versatility afforded by the modular assembly and wide range of configurations of the rapid-prototyped printheads allow for the fabrication of microfibers with diverse compositions and geometries (FIG. 13A). Applications for use of this platform include printing multiple cells, drug-releasing meshes, and sensors.
  • the bioprinters include high-resolution stereolithography (SLA) 3D printing and microfluidic technologies to develop an inexpensive, modular handheld bioprinting system (FIGS. 1A-12I).
  • the device is composed of (i) a printhead made with a high- resolution resin SLA 3D printer, (ii) pneumatic syringe cartridges adapted for hydraulic extrusion, (iii) a 3D printed cartridge enclosure, (iv) a mountable photocrosslinking system composed of a 3D printed case with an array of inexpensive 3mm 405nm LEDs (light emitting diodes), and (v) an array of external syringe pumps for hydraulic material extrusion, including different cross-sectional shaped fibers, grooved fibers, hollow fibers, multi-component fibers with material gradients along the fiber length or cross-section, co-axial, tri-axial and multiple-core fibers.
  • SLA stereolithography
  • FIGS. 1-10 the device is composed of (i) a printhead made with a high
  • the bioprinter platform also introduces a hydraulic-driven extrusion mechanism.
  • This mechanism was chosen over current handheld extrusion mechanisms, such as pressure driven extrusion since the flowrate can be directly set on the syringe pump and remains constant regardless of the viscosity and temperature of the bioink, eliminating the need to adjust pressure values for different bioinks and temperature variations, which can be an arduous process, especially for low viscosity materials.
  • having the syringe pumps separate from the device significantly reduces its size and weight compared to direct motor driven extrusion mechanisms, resulting in a more ergonomic and comfortable design for improved long-term operation, while allowing for the addition of multiple materials while maintaining a compact form factor.
  • bioprinters can incorporate drug carriers, as well as biochemical and biophysical cues to demonstrate the versatility of the handheld bioprinter in developing spatiotemporal drug delivery modules, as well as wearable biosensors. Additionally, the proof of principle demonstration for the in situ printing of single-core and dual core cell-laden fibers were illustrated to further extend the versatility of this platform in various biomedical applications.
  • the bioprinted fiber can carry biologically active molecules such as drugs, growth factors and proteins for local delivery to the injury site to promote healing rate.
  • the bioprinter system can be tailored to be used as a platform for the temporal and dosage-control delivery of different biomolecules (FIG. 13C). This feature allows precise control over the release rate as well as deliver multiple agents during the healing period. This was achieved by printing fibers made of poly(ethylene glycol) diacrylate (PEGDA)-Laponite carrying different concentrations of PLGA particles (3, 6, 9 mg/ml), which were loaded with Rhodamine B (Rd) or 40 kDa fluorescein isothiocyanate-dextran (FITC-Dextran). The single-channel printhead was used to print both fiber types onto a petri dish, followed by photocrosslinking by illuminating 405 nm light at 20 mW/cm 2 for 5 minutes,
  • PNIPAM Poly(N-isopropylacrylamide)
  • PEGDA-Laponite fibers loaded with PNIPAM microcarriers were printed.
  • the PNIPAM microcarriers were produced and loaded with FITC-Dextran.
  • the FITC-Dextran release profile demonstrated a temperature -responsive behavior in which an increase in temperature from 20 to 42 °C resulted in significantly higher diffusive flux of FITC-Dextran into the release medium.
  • electroconductive materials can be incorporated in the printed fibers, which can be used as an electronic system for facilitating heattrigger drug release by altering the temperature. Equipping this temperature-responsive drug delivery composition with an electronic system allows for activation of drug release from each individual fiber, providing precise control of the drug dosage by triggering an adequate quantity of drug-eluting fibers.
  • the release profile of Rd-loaded PLGA carriers from the printed fiber showed a linear diffusive release to the level of 21.1 ⁇ 3.2 during the release period.
  • the release rate of Rd reached to significantly higher levels compared to BSA and FITC-Dextran, owing to the smaller size of Rd, which facilitates its flux from the pores presented in the PLGA microcarriers and oriented fiber.
  • a dual-material (SBS) printhead was utilized as shown in FIG. 15A.
  • fibroblast and HaCaT cells were suspended each in a separate bioink composed of 1, 5, and 0.3% (w/v) alginate, GelMA, and lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), respectively.
  • a monoculture fiber configuration was made by feeding fibroblast-loaded bioink into both channels of the printhead.
  • a co-culture fiber configuration was made by feeding fibroblast-loaded bioink through one channel, and HaCaT -loaded bioink through the other channel.
  • Both channels merged into a single one right before reaching the tip of the printhead.
  • the cell-laden fibers were wet spun into CaCL 1% (w/v) in DPBS solution, followed by photocrosslinking by illuminating 405 nm light at 20 mW/cm 2 for 5 minutes.
  • the encapsulated cells in the fibers were visualized and distinguished via staining nuclei with DAPI and mesenchymal cells with vimentin (FIGS. 15B- 15C).
  • the confocal images showed that after 7 days of culture, both cell types populated across the fiber.
  • FIG. 15B shows that the fibroblast cells (stained positive with DAPI and Vimentin) completely covered the fiber after 7 days of culture in the mono-culture configuration.
  • FIG. 15B shows that the fibroblast cells (stained positive with DAPI and Vimentin) completely covered the fiber after 7 days of culture in the mono-culture configuration.
  • FIG. 15B shows that the fibroblast cells (stained positive with
  • 15C shows that in the co-culture configuration, clusters of HaCaT cells (stained positive with DAPI) were formed in the right half of the fiber, while the fibroblast cells not only spread across the left half of fiber, but also partially infiltrated into the right half.
  • Post-fabrication live/dead images of the produced fibers revealed that the majority of cells stained with calcein (live) implying that the fabrication technique had small effect on the cell viability.
  • the ultrafast, visible light photocrosslinking approach offered by the bioprinters disclosed herein can reduce cell death caused by UV exposure.
  • Photocrosslinkable polymers such as methacrylated hyaluronic acid (HA-MA) [31] and GelMA, as well as polyethylene glycol (PEG) derivates like PEGDA hold a great potential to be used in fabricating three-dimensional cell-laden tissue models.
  • HA-MA methacrylated hyaluronic acid
  • PEG polyethylene glycol
  • the constructs made of photocrosslinkable polymers have been shown to have high printing resolution.
  • the spatiotemporal control along with the low-temperature production mode offered by these photocrosslinkable bioinks facilitate 3D printing of complex cell-laden structures.
  • cell death caused by UV light irradiation and cytotoxicity induced by the photo initiator (PI) is a challenge associated with 3D-printing of cell-laden photocrosslinkable polymers.
  • the bioprinters disclosed herein offer an ultrafast photocrosslinking approach using a low-cost LED sleeve.
  • 3 million C2C12 cells were suspended in a solution composed of 10% GelMA solution with 0.3% LAP and printed into rectilinear scaffolds.
  • the nuclei and actin microfilaments of C2C12 cells encapsulated in the printed construct were visualized by DAPI/actin staining.
  • the confocal image taken after one day showed an even 3D distribution of cell throughout the fiber (FIG. 16C).
  • FIG. 16C shows an even 3D distribution of cell throughout the fiber.
  • FIG. 16F shows that the printed constructs were almost confluent with C2C12 cells after 8 days of incubation (whereas FIG. 16E shows one day of incubation). Besides, the results demonstrated that many of C2C12 cells developed a myotubule morphology by day 8.
  • the proliferation rate of C2C12 encapsulated in the fiber were quantified by measuring the metabolic activity of cells. As shown in FIG. 161, C2C12 cells proliferated over the 2 days of incubation. The metabolic activity in the second day of culture was twice as much as compared to the first day.
  • CM-encapsulated fibers were assessed for functional properties by immunofluorescence staining of DAPI/Actin/Troponin and beating properties of GelMA bioink.
  • the printed constructs showed a high level of Troponin T expression, showing a high level of live CMs present within the fiber.
  • CM-laden GelMA microfibers showed elongated and rod-shaped morphology of structural protein (F-actin) and contractile protein (Troponin-T), demonstrating relevant CM phenotype (FIGS. 16G- 16H). This was further indicated by the beating of cells leading to the movement of fibers after 5 days of culture.
  • GelMA microfibers provided a favorable microenvironment for the growth, proliferation, and function of CMs.
  • CMs were encapsulated in the bioink and printed in the same coaxial configuration.
  • a live-dead assay also showed great cytocompatibility of the bioink after 7 days of culture (FIGS. 17B, 17F-17H).
  • the handheld bioprinter was also used to produce hydrogel fibers with multiple cores, which enables modelling the biological behavior of cancer cells in vitro.
  • the bioprinter used a double-core fiber composed of GelMA 5% (w/v) in each core channel, and a matrix of GelMA 5% (w/v)/ alginate 1% (w/v) in the sheath channel to model the invasion of the ovarian cancer cells into the adjacent ECM.
  • tumor models with SKOV-3 cells were loaded into one of the cores and human-derived fibroblast cells were loaded into the adjacent core creating a co-culture fiber model.
  • a mono-culture model was made by loading SKOV-3 cells into a single core, while leaving the adjacent core without cells.
  • the fluorescent images of the tumor and stromal cells inside the fibers demonstrated a time-dependent invasion pattern of the tumor cells from the GelMA core of the fiber into the matrix.
  • the invasion length of the cancer cells was quantified by measuring the migrated cells to the farthest point of the matrix in the longitudinal direction of the fibers. Results showed that in both mono and co-culture conditions, SKOV-3 cell invasion length increased over time from day 1 to day 3. It should be noted that there was a significant difference in invasion length of the cells into the matrix when they were co-cultured with fibroblast cells.
  • the wet spun fibers can be considered as an in vitro tumor model for recapitulating the micro physiological environment of in vivo tumor. Metabolic activity analysis of the cells in both mono- and co-culture conditions were in line with Eive/Dead images of the cells, illustrating the viability of both cells in the fiber spinning and culture condition over time.
  • Cancer and fibroblast cells in the fibers were shown to proliferate for at least 3 day such that the presence of the fibroblasts within the fibers caused a significant increase in normalized fluorescent intensity index of the cells. This suggests a higher metabolic activity of the cells in co-culture in comparison to the mono-culture condition.
  • Vascularized tissue is one of the main challenges to engineer biomimicking artificial tissue to restore or replace a damaged tissue.
  • Hollow fibers architectures facilitate transportation of nutrition into the engineered organ and removal of waste to promote healing rate.
  • the handheld bioprinters disclosed herein enabled the convenient manual deposition of perfusable fibers to generate constructs in two (FIG. 19B) or three dimensions FIGS. 19C and 19F).
  • Coaxial and dualcore printheads were used to demonstrate the feasibility to fabricate single channel (FIGS. 19B- 19F) or multichannel (FIG. 19G) hollow, perfusable vascular structures with consistent or varying diameters.
  • an aqueous solution composed of alginate 2% (w/v) and Eaponite 6% (w/v) bioink was flowed through the sheath channel while a solution of PVA 10% (w/v) and CaCh 1 % (w/v) ran through the core channel(s) to induce ionic crosslinking of the alginate.
  • Eaponite was used in the alginate bioink to maintain structural integrity during fiber crosslinking.
  • the outer and inner fiber diameters can be tuned by varying the core and sheath flowrates, as well as the printing speed (FIG. 19D).
  • pH is one of the indicators of physiological condition of skin and can be served as an effective real-time monitoring system.
  • Color-changing pH sensor array can be used for early detection of bacterial infections in wounds using image processing applications and smartphones.
  • beads carrying colorimetric pH-sensitive dye are incorporated in alginate 2% (w/v)/Laponite 6% (w/v).
  • FIG. 20 A demonstrates direct deposition of pH-responsive colorimetric sensor on the generated artificial pig skin. As shown by the photographic image of the sensors exposed to buffers with different pH, the color of sensors changed from colorless to dark blue in response to variations in pH within the range of 7-8.75 (FIG. 20B).
  • This method can be served as a point-of-care device to continuously monitor pH level of the defect area and examine the healing process.
  • graphene oxide has been added to the starting alginate/laponite bioink to induce electro conductivity to the printed fiber.
  • This feature is widely applicable in the development of biosensors, wearable or implantable bioelectronics, and heating element to trigger on-demand temperature -responsive drug release.
  • electroconductive hydrogels are versatile platforms applicable in engineering neural and cardiac tissue. This is due to the fact that the stimulation of cellular behavior (i.e. differentiation, migration, and proliferation) can be done via applying electrical, electrochemical, and electromechanical signals.
  • FIGS. 20C-20D show the capability of the handheld printer to produce electroconductive construct directly at a site of injury. As shown by the photographic image of FIG. 20C, two conductive fibers printed on the pig skin wound were capable of illuminating a small LED when a voltage was applied from an external power supply.
  • the bioprinters and bioprinter systems disclosed herein can be a low-cost handheld bioprinting platform, that enables in situ deposition of functional hydrogel fibers and different types of cells such as dermal fibroblasts, HaCaT, C2C12, and SKOV-3 cells.
  • the printheads can be fabricated with a high-resolution SLA-DLP 3D printer.
  • the modularity of the platform can allow for the equipment of a mountable low-cost photocrosslinking system and pneumatic Luer lock cartridges, as well as facilitating easy and rapid modification of the device for different bioink compositions and printing geometries.
  • the bioprinters and bioprinter systems disclosed herein can enable the direct extrusion of fibers with different morphological characteristics, size, cross-sections, and material gradients at the site of injury. Additionally, the bioprinters and bioprinter systems disclosed herein can be capable of printing fibers with a wide range of materials such as alginate, GelMA, and PEGDA in the pure or hybrid forms using different crosslinking approach.
  • the printed fibers can have the versatility to be augmented with drug carriers, conductive materials, and pH responsive sensors, which extend the application of the printed fibers to the developing spatial and temporal local drug delivery systems, tissue engineering scaffolds, and wearable biosensors and bioelectronics.
  • having multichannel cartridge design can allow to print multicore cell laden fibers in which different types of cells are co-cultured and proliferated, showing the feasibility of the bioprinters for the direct cell delivery applications.
  • This ability to produce dual core cell laden hydrogel fiber provide the opportunity to study biological behavior of cancer cells in vitro. Taking everything in consideration, in the context of regenerative medicine, the bioprinters and bioprinter systems disclosed herein can be used in clinical settings for efficient delivery of cells and therapeutic agents.
  • Polylactic acid (PLA), Gelatin Type A from porcine skin, Methacrylic anhydride (MA), LAP, alginate, Poly(ethylene glycol) diacrylate (PEGDA, Mn 2000 Da), graphene oxide, Poly(vinyl alcohol) (PVA), Hank’s balanced salt solution, Rhodamine B (Rd), bovine serum albumin (BSA), 20 KDa fluorescein isothiocyanate- dextran solution (FITC- Dextran), dichloromethane (DCM), poly(lactic-co-glycolic acid) (PLGA), Poly(N- isopropylacrylamide) (NIP AM), N,N'-methylenebis (acrylamide) (BIS), 2-hydroxy-4'-(2- hydroxyethoxy)-2-methylpropiophenone as photoinitiator (PI), ammonium persulfate (APS), Span 80, and PBS tablets were purchased from Sigma- Aldrich.
  • 3-ml bioink cartridges were purchased from Cellink (BICO, Sweden). To fit within the compact design of the device, the barrel flanges were cut with an exacto knife and replaced with the 3D-printed barbed connectors. Individual syringe pumps (Harvard Apparatus) with water-loaded syringes were connected to the barbed ends of the cartridges with plastic tubing for hydraulic actuation powered extrusion. The bioink cartridges were attached to the printhead with female-to-female Luer lock connectors. Six 3-mm 405nm LEDs (Mouser Electronics, Canada) and a push button were embedded to the 3D-printed light curing case and connected in series, powered by two 9V batteries to supply 3V to each LED.
  • components of the handheld bioprinter were designed using Fusion 360 software (Autodesk, Inc.).
  • the housing and LED case were printed in PLA material on an Ultimaker 2 3D printer.
  • the printheads and barbed attachments were printed in photocurable resin using a Kudo3D Micro SLA 3D printer.
  • solid GelMA was prepared using type-A gelatin from porcine skin (Sigma- Aldrich) was dissolved in PBS (ThermoFisher) at a concentration of 5% (w/v) at 60 °C .
  • Methacrylic anhydride (MA) was slowly added to the gelatin solution while stirring at 300 rpm. The solution was left stirring for 3 h and transferred into 12-14 kDa dialysis membranes. The membranes were placed in a large beaker with deionized water at 50 °C. The water was replaced twice a day for 7 days to remove unreacted MA. The solution was then filtered and lyophilized for 3 days to obtain solid GelMA.
  • GelMA bioink was made by dissolving solid GelMA in Dulbecco's Modified Eagle's Medium (DMEM, ThermoFisher Scientific, cat# 11965084) mixed with lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) (Sigma-Aldrich). GelMA and LAP concentrations varied for different tests. The final solution was filtered with a 0.2 um sterile filter.
  • DMEM Dulbecco's Modified Eagle's Medium
  • LAP lithium phenyl-2,4,6-trimethylbenzoylphosphinate
  • an alginate stock solution was made by dissolving sodium alginate powder (Sigma- Aldrich) in DI water at a concentration of 4% (w/v) and kept at 60°C for 1 h while stirring. The solution was then transferred into a syringe.
  • Laponite stock solution was made by adding Laponite powder (BYK, Germany) to a falcon tube with DI water at a concentration of 12% (w/v) at 4°C to prevent immediate gelation, and vortexed for 2 min. The contents were immediately transferred into a syringe for the solution to gelate without creating bubbles.
  • Both alginate and laponite stock solutions were mixed at a 50:50 ratio by connecting both syringes with a female-to-female Luer lock adapter and repeatedly pushing the material back and forth between both syringes, resulting in a homogeneous solution with final concentrations of 2% alginate and 6% Laponite.
  • PEGDA bioink was made by dissolving LAP in distilled water at 60°C at a concentration of 0.3% (w/v). The solution was left in a water bath at 60°C for 15 min, periodically vortexing every 5 min. PEGDA (Sigma- Aldrich) was then added to the solution to a concentration of 7.5% (v/v) and vortexed for 2 min. The solution was then transferred into a syringe.
  • a PEGDA stock solution was made by dissolving LAP in distilled water at 60°C at a concentration of 0.6% (w/v). The solution was left in a water bath at 60°C for 15 min, periodically vortexing every 5 min. PEGDA (Sigma- Aldrich) was then added to the solution to a concentration of 15% (v/v) and vortexed for 2 min. The solution was then transferred into a syringe. Separately, a 12% (w/v) Laponite stock solution was prepared using the same method described in the “alginate/Laponite” bioink section above.
  • alginate powder was sterilized by suspending it in anhydrous ethanol in a 15 ml Falcon tube, and left without a lid inside a BSC until the ethanol was fully evaporated.
  • An alginate stock solution was prepared by dissolving the sterilized powder in DMEM at 2% (w/v).
  • a separate stock solution of 10% (w/v) GelMA and 0.6% (w/v) LAP was prepared in DMEM and filtered with a 0.2 um acetate cellulose syringe filter. Both GelMA and alginate stock solutions were mixed at a 50:50 ratio by connecting both syringes with a female-to-female Luer lock adapter and repeatedly pushing the material back and forth between both syringes, resulting in a homogeneous solution with final concentrations of 5% (w/v) GelMA, 0.3% (w/v) LAP and 1% (w/v) alginate.
  • pH responsive bioink was prepared from about 135 mg of a- naphtholphthalein dye (Sigma- Aldrich) dissolved in a beaker with 6 mL of 100% ethanol, followed by the addition of 24 mL dH2O and stirred for 30 m. 3300 mg of Dowex 1 x 4 chloride form (Sigma- Aldrich) beads were washed in a 50 mL Falcon tube with DI water by vortexing for 2 min, letting the beads fully sediment and replacing the supernatant. This washing step was repeated twice with DI water and once with 100% ethanol.
  • a- naphtholphthalein dye Sigma- Aldrich
  • graphene oxide bioink was prepared using graphene oxide (Sigma- Aldrich) was DI water at a concentration of 5 mg/ml and sonicated for 5 min. Alginate powder was added to the suspension at a concentration of 0.5% (w/v) and vortexed for 5 min. The solution was cooled to 4°C. Laponite powder was then added at a concentration of 6% (w/v) and vortexed for 2 min. The contents were immediately transferred into a syringe for the solution to gelate without creating bubbles.
  • the desired cells were dissociated, centrifuged, and the entire supernatant was carefully removed. Using a pipette with the tip cut, the desired sterile bioink was added to the cell pellet which was resuspended by gentle pipetting.
  • PVA Sigma- Aldrich
  • CaCh powder Micros Organics
  • the drug models were initially loaded on the PLGA microcarriers.
  • the double (W1/0/W2) emulsion/solvent evaporation technique was used to prepare drug- loaded PLGA microparticles.
  • 1 ml PVA 1% (w/v) aqueous solution containing 1 mg of the drug (Rd, FITC-Dextran or bovine serum albumin (BSA, Sigma Aldrich, Cat# A3803)) (Wl) was emulsified into 5 ml of dichloromethane (DCM) solution containing 10% (w/v) PLGA and vortexed for 1 min to prepare the primary Wl/O emulsion.
  • DCM dichloromethane
  • PNIPAM microcarriers prepared according to the previously published method with the assist of microfluidic flow-focusing droplet generators. Briefly, the dispersed solution was prepared by dissolving PNIPAM (10%, w/v), 2-hydroxy-l-(4-(hydroxy ethoxy) phenyl)-2-methyl-l -propanone N,N'-methylenebis (acrylamide) (0.4%, w/v), 2-hydroxy-4'-(2- hydroxyethoxy)-2-methylpropiophenone PI (1% w/v), and ammonium persulfate (APS) (0.6%, w/v) in distilled water.
  • the dispersed phase and continuous phase composed of mineral oil and span 80 (10%, v/v) were injected into a microfluidic chip using 1 ml and 10 ml syringes which were connected to the inlets of chip with Tygon tubings. These syringes were mounted on syringe pumps (Harvard Apparatus PHD 2000, USA) to precisely control the flow rates of continuous and dispersed phases. In situ photopolymerization of the microcarriers was carried out when the generated microcarriers passed through a coil-shaped collecting tube while being exposed to UV light (20 mW/ cm-2) for 15 minutes.
  • FITC- Dextran was passively loaded onto the microparticles by immersing 10 mg of the lyophilized particles into a 1ml of FITC-Dextran solution (1 mg/ml) for 48 hours. The suspension vortexed every 12 hours to ensure that FITC-Dextran uniformly loaded on the carriers. Then, the suspension was centrifuged to remove FITC-dextran solution followed by washing the particles 3 times with cold water (15 °C).
  • the extrusion rate of each channel was individually adjusted by setting the flowrate of their respective syringe pumps to the desired value. Bioprinting started as soon as all the materials reached the tip of the printhead. The printing surface or wet spinning media varied for each test. It should be noted that for all cell work, the interiors of the printhead, hydrogel cartridges, connectors, and tubing were all sterilized with 70% ethanol and washed twice with DPBS prior to printing the cells. Also, the exterior of all components were sprayed with 70% ethanol. The tubing and each cartridge were then filled with DPBS and a syringe with DPBS was connected to a syringe pump for hydraulic actuation of each cartridge.
  • alginate-laponite bioink was loaded into one cartridge and attached to the sheath channel of the coaxial printhead.
  • PVA-CaCh sacrificial crosslinking solution was loaded into a separate cartridge and attached to the core channel of the printhead.
  • two cartridges with PVA-CaCh were attached to each core channel of the dual-core printhead.
  • the fibers were printed on a petri dish or an artificially made pig skin wound.
  • CaCh (1% w/v) was deposited over the fibers immediately after printing to accelerate crosslinking.
  • the sacrificial PVA- CaCL solution was cleared from the internal fiber channel using an air-filled syringe with a 25G needle inserted at the end of the fiber. The fiber was then perfused with dye using the same syringe and needle.
  • drug-loaded PLGA microcarriers were dispersed into PEGDA-laponite bioink at concentrations of 3, 6, and 9 mg/ml.
  • Each drugeluting fiber was made by extruding 60 pl of the PLGA-loaded bioink followed by photopolymerizing for 5 minutes under 405nm visible light.
  • the fiber loaded with PLGA drug carriers was immersed in 300 pl of PBS and incubated at 37 °C. At predefined timepoints, 100 pl of the release medium sample was collected to measure the drug concentration and replaced with 100 pl of fresh PBS.
  • the fluorescence intensity of the collected medium at each time point was measured using a Tecan Infinite M Nano plate reader at lex 490 nm / kern 520 nm for FITC dextran, and lex 546 nm / kern 585 nm for Rh. Using the relevant calibration curve, the acquired fluorescence intensity was converted to concentration (mg/ml).
  • PLGA microcarriers loaded with bovine serum albumin (BSA), FITC- Dextran and Rh were dispersed into three separate PEGD A bioinks at a concentration of 18 mg/ml (PLGA/bioink). All three bioinks were loaded into three separate cartridges. The cartridges were attached to each channel of the 3-material (SBS) printhead with the LED module attached. The 3- material drug-eluting fibers were made by extruding all bioinks at the same flowrate until a total of 60 pl was extruded. The resulting fiber had a concentration of 6 mg/ml of each drug-loaded microcarrier.
  • BSA bovine serum albumin
  • FITC- Dextran FITC- Dextran and Rh
  • FITC dextran, Rh, and BSA 3 drugs
  • 60 pL of drug-loaded fiber was incubated in 300 pL of PBS.
  • 100 pL of the medium sample was obtained to test the drug concentration at predefined time intervals, and 100 pL of fresh PBS was supplied to maintain a constant volume of release medium.
  • the fluorescence intensity of the withdrawn medium at each time point was measured using a Tecan Infinite M Nano plate reader to determine the release of FITC dextran and Rh (lex 490 nm / kern 520 nm for FITC dextran, and lex 546 nm / kern 585 nm for Rh).
  • the acquired fluorescence intensity was converted to concentration (mg/mL).
  • concentration mg/mL
  • 500 pL of Bradford Reagent was added to 10 pL of the sample, and the absorbance was measured at 595 nm using the plate reader, and the absorbance was then connected to the concentration of BSA using a standard curve.
  • the drug release percentage at various time periods was calculated. Six replicates were used in each drug release experiment.
  • thermoresponsive, drug-eluting PNIPAM microcarriers were dispersed into PEGDA-Laponite bioink at a concentration of 10 mg/ml.
  • Each drug-eluting fiber was made by extruding 60 pl of the blended ink followed by photopolymerizing for 5 minutes under 405nm visible light. After that, the release experiment was carried out by immersing the prepared fibers in 300 pl of PBS at 20, 37, 42 °C in pH 7.4. At predefined time intervals, 100 pl of the supernatant was taken for analysis and replaced with fresh PBS with temperature similar to that of release condition.
  • the concentration of FITC-Dextran in the supernatant was analyzed by measuring the fluorescence intensity at an excitation peak of 490 nm and an emission peak of 520 nm using a plate reader (Tecan Infinite M200Pro).
  • the cumulative FITC-Dextran release from fibers was calculated via converting fluorescence intensity to concentration by the prepared standard calibration curve. This experiment was performed in triplicate.
  • pH responsive colorimetric bioink was loaded into a cartridge and attached to the single-channel printhead.
  • 2-layer meshes were printed directly on an artificially made pig skin wound, and crosslinked by adding 1-ml of CaCb 1% (w/v) solution to each fiber and let sit for 3 minutes. The excess CaCb was removed with a Kimwipe (Kimberly-Clark Worldwide, Inc.). 200 pl of buffer solutions at different pH values (7, 7.5, 8, 8.75) were added to each mesh and let sit for 1 h for colorimetric response to take effect.
  • graphene oxide bioink was loaded into a cartridge and attached to the single-channel printhead. Two fibers were printed directly on an artificially made pig skin wound, and crosslinked by adding 1-ml of CaCh 1% (w/v) solution to each fiber and let sit for 3 minutes.
  • the excess CaCh was removed with a Kimwipe and the fibers and underlying pig skin were washed twice with DI water.
  • two separate sterile alginate-GelMA bioink solutions were loaded with HaCaT cells and fibroblasts, respectively.
  • Two cartridges were loaded with the fibroblast-laden bioink, and one cartridge was loaded with HaCaT cell-laden bioink.
  • the two fibroblast-laden cartridges were attached to either side of the 2-material (SBS) printhead.
  • SBS 2-material
  • one fibroblast-laden cartridge was attached to one channel and the HaCaT cell-laden cartridge was attached to the other.
  • sterile GelMA 10% (w/v) + LAP 0.3% bioink solution was loaded with C2C12 myoblasts cells (ATCC CRL-1772) (3 x 10 6 cell/ mL) or CMs (15 x 10 6 cell/ mL).
  • the cartridge was cooled down to 15°C For thermal gelation of GelMA prior to printing.
  • the LED module and cooling sleeve (stored at 4°C) were mounted to the printhead. Fibers or meshes were printed on a petri dish inside the BSC while simultaneously being photocrosslinked with the LED module. The constructs were then transferred to culture media.
  • the in vitro tumor model was made the wet spinning of dual-core fibers including 2 compartments for tumor and stroma.
  • Alginate/GelMA bioink was loaded into one cartridge and attached to the sheath channel of the dual-core printhead.
  • Three separate sterile GelMA 5% (w/v) + LAP 0.3% (w/v) solutions were loaded with either fibroblast cells (2 x 10 6 cell/ mL), human ovarian cancer cell line (Skov-3) (4 x 10 6 cell/mL), or left without cells (blank). All three solutions were loaded into three separate cartridges.
  • human neonatal dermal fibroblasts (ATCC PCS-201-010) were cultured in high glucose DMEM supplemented with 10% fetal bovine serum (FBS, ThermoFisher Scientific, cat# 10437028) and 1% (v/v) 1% penicillin-streptomycin (Pen Strep) (Cat: 15140122, Invitrogen/Eife Technologies) in an atmosphere of 5% CO2 at 37 °C. cells were cultured in a in Corning T-75 cm 2 Rectangular Canted Neck Cell Culture Flask and passaged with TrypLE Express (ThermoFisher Scientific, cat# 12605093) once they got around 90% confluence.
  • FBS fetal bovine serum
  • Pen Strep penicillin-streptomycin
  • HaCaT cells (AddexBio, T0020001), a transformed human keratinocyte cell line, were cultured in high glucose DMEM with 10% FBS in Corning T-75 cm 2 Rectangular Canted Neck Cell Culture Flask and passaged with Trypsin-EDTA (ThermoFisher, cat# 25200072).
  • the culturing protocol for C2C12 myoblasts cells (ATCC CRE-1772) and human ovarian cancer cell line Skov-3 cells (HTB-77) was the same as the protocol for HaCaT cells.
  • NIH-3T3 fibroblasts were cultured in a medium composed of DMEM supplemented with 10% CBS and 1% antibiotic in a sterile incubator (Heracell Vios 160i, Thermo Scientific, US) with 5% CO2 at 37 °C. The cells were passaged after reaching around 80% confluence.
  • neonatal Sprague-Dawley rats (2 days old) were obtained from Charles River Laboratories (Canada) and were euthanized based on the protocol approved by the Animal Ethics Committee of the Centre Hospitalier Universitaire Sainte -Justine Research Centre (CRCHUSJ). All the procedures were in accordance with the guidelines of the Canadian Council on Animal Care and the US National Institutes of Health Guide for the Care and Use of Laboratory Animals. The hearts were collected into HBSS cold buffer, cut into smaller sections, washed with cold HBSS two times, and digested to single cardiac cells by the Neonatal Heart Dissociation Kit (Miltenyi Biotec, Germany) based on the recommended protocol of the manufacturer.
  • CRCHUSJ Animal Ethics Committee of the Centre Hospitalier Universitaire Sainte -Justine Research Centre
  • CMs were isolated from cardiac fibroblasts (CFs) via centrifugation as a cell pellet.
  • printed samples were transferred to the 6 well-plate and cultured containing the specific cell growth media in an atmosphere of 5% CO2 at 37 °C.
  • a live/dead cell viability kit (Invitrogen, L3224) was used following the manufacturer's instructions.
  • the fibroblast, HaCaT, and Skov-3 cell-laden fibers or meshes were incubated in the live/dead solution for 30 minutes while protected from light at room temperature.
  • the hydrogels were then washed once with DPBS and imaged with a Zeiss Axio Observer 5 fluorescent microscope (Zeiss, Germany).
  • CM-laden fibers the fibers were washed with HBSS three times and were further imaged by either Leica DMi8 wide-field or SP8-DLS confocal microscopes (Leica Biosystems, Germany).
  • fibers were fixed with 10% neutral buffered formalin (ThermoFisher Scientific, Cat# 22-220682) for 45 minutes at room temperature then washed 3 times with PBS.
  • the fibers were permeabilized with 0.3% Triton- Xi 00 (Sigma Aldrich, Cat# XI 00) in PBS for 10 minutes, then incubated in a blocking buffer of 3% BSA and 0.3% Triton-XlOO in PBS for 30 minutes.
  • the primary antibody solutions were prepared in 1% BSA and 0.3% Triton-XlOO in PBS with a dilution of 1:200 for anti-vimentin conjugated to AlexaFluor 488 (source, Cat#). Hydrogel fibers were incubated in the primary antibody solution overnight at 4°C protected from light.
  • the fibers were then washed three times with PBS and the nuclei were counterstained with 5pg/ml 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI, Sigma Aldrich, Cat# D9542) for 10 minutes.
  • DAPI 5pg/ml 4',6-Diamidino-2-phenylindole dihydrochloride
  • the fibers were again washed three times with DPBS and finally imaged with a Zeiss Axio Observer 5 fluorescent microscope.
  • CM-laden fibers after 5 days of culture, CM-laden fibers were fixed into 3.7% formaldehyde solution for 1 h at room temperature, followed by washing 3 times with HBSS. The fibers were permeabilized with 0.3% Triton-X 100 in HBSS for 30 minutes and then placed into a blocking buffer of 5% BSA and 0.3% Triton-X 100 solution for 1 h. Cardiac troponin T (cTnT) primary antibody solution was prepared by dilution into 1% BSA solution (1:200), and the fibers were incubated in this solution at 4 °C overnight.
  • cTnT Cardiac troponin T
  • the fibers were further placed into 1% BSA solution containing donkey anti-rabbit- Alexa Fluor 488 secondary antibody (1:400) and Phalloidin iFlour 594 (1:1000) for 1 h. After three times washing with HBSS, the counterstaining for nuclei was performed with DAPI solution (1:1000) for 10 minutes. The fibers were washed 3 times with HBSS, and confocal imaging was done with SP8-DLS Leica microscope.
  • a PrestoblueTM assay (Invitrogen, A13261), was used to evaluate the proliferation of cells over time.
  • Cell-laden fibers were cut into 5 mm sections with a razor blade which were carefully transferred into a 96 well plate where they were cultured in complete media. At each timepoint, the fiber sections were transferred to fresh wells each containing 110 pL Prestoblue working solution (10% (v/v) Prestoblue reagent in media). After a 1-hour incubation at 37°C, 100 pL was transferred from each well to a separate plate and read on a microplate reader (560/590 nm ex/em). Cell-laden fibers were washed with media then returned to culture. In some examples, after 5 days of culture, the beating of CM-encapsulated fibers was recorded by EVOS M5000 digital inverted microscope (Thermo Fisher Scientific, USA).
  • a bioprinting system comprising: a handpiece comprising: a printhead configured to have one or more bioink cartridges fluidly coupled thereto; a temperature control module comprising a channel configured to receive the one or more bioink cartridges therein; and a syringe pump subsystem comprising one or more syringe pumps, each of the one or more syringe pumps configured for fluid communication with one of the one or more bioink cartridges and configured to control flow of bioink from the respective bioink cartridge.
  • Example 2 The bioprinting system of any example disclosed herein, particularly example 1 , wherein the handpiece further comprises a light-emitting device configured to be attached to the printhead, the light-emitting device comprising one or a plurality of light sources configured to emit light at a selected wavelength for photo-crosslinking of bioink.
  • a light-emitting device configured to be attached to the printhead, the light-emitting device comprising one or a plurality of light sources configured to emit light at a selected wavelength for photo-crosslinking of bioink.
  • Example 3 The bioprinting system of any example disclosed herein, particularly example 1, wherein the temperature control module further comprises an interior wall defining the channel, an exterior wall, a chamber defined between the interior wall and the exterior wall, and a phase change material disposed within the chamber.
  • Example 4 The bioprinting system of any example disclosed herein, particularly example 1 , further comprising a computerized controller comprising at least one processor and non- transitory memory storing one or more programs, the computerized controller in data communication with the pump subsystem, the one or more programs configured to, when executed by the at least one processor, cause the computerized controller to, control a respective pressure or flow rate setting for each of the one or more syringe pumps based at least on data related to a type of bioink in each of the respective one or more bioink cartridges.
  • a computerized controller comprising at least one processor and non- transitory memory storing one or more programs, the computerized controller in data communication with the pump subsystem, the one or more programs configured to, when executed by the at least one processor, cause the computerized controller to, control a respective pressure or flow rate setting for each of the one or more syringe pumps based at least on data related to a type of bioink in each of the respective one or more bioink cartridges.
  • Example 5 The bioprinting system of any example disclosed herein, particularly example
  • a sensor configured to detect one or more of flow of bioink, a position of bioink, or a structure of bioink dispensed from the printhead, the sensor in signal communication with the computerized controller and configured to provide data related to bioink flow, bioink position, and/or bioink structure thereto.
  • Example 6 The bioprinting system of any example disclosed herein, particularly example
  • the computerized controller is further configured to, based at least in part on the data related to bioink flow, bioink position, and/or bioink structure indicating insufficient dispensement of bioink for one of the one or more bioink cartridges, increase the respective pressure or flow rate setting for the syringe pump in fluid communication with the one of the one or more bioink cartridges.
  • Example 7 The bioprinting system of any example disclosed herein, particularly example 5, wherein the computerized controller is further configured to, based at least in part on the data related to bioink flow, bioink position, and/or bioink structure indicating over dispensement of bioink for one of the one or more bioink cartridges, decrease the respective pressure or flow rate setting for the syringe pump in fluid communication with the one of the one or more bioink cartridges.
  • Example 8 The bioprinting system of any example disclosed herein, particularly example 1 , wherein the handpiece further comprises one or more actuators configured to control flow of bioink from each of the one or more bioink cartridges.
  • Example 9 The bioprinting system of any example disclosed herein, particularly example 1 , wherein the printhead is a multichannel printhead configured to have three or more bioink cartridges fluidly coupled thereto.
  • Example 10 The bioprinting system of any example disclosed herein, particularly example 1 , wherein the syringe pump system comprises a syringe pump array comprising one or more hydraulic syringe pumps configured for hydraulic bioink extrusion.
  • Example 11 A handheld bioprinter comprising: a multi-channel printhead configured to have three or more bioink cartridges fluidly coupled thereto; and a housing configured to receive the three or more bioink cartridges when coupled to the multi-channel printhead.
  • Example 12 The handheld bioprinter of any example disclosed herein, particularly example 11 , wherein the housing comprises a temperature control module comprising a central channel configured to receive the three or more bioink cartridges.
  • Example 13 The handheld bioprinter of any example disclosed herein, particularly example 11, further comprising three or more fluid couplings, each of the three or more fluid couplings configured to fluidly couple one of the three more bioink cartridges with an external syringe pump array comprising three or more syringe pumps, each of the three or more syringe pumps configured for fluid communication with one of the three or more bioink cartridges and configured to control flow of bioink from the respective bioink cartridge.
  • Example 14 The handheld bioprinter of any example disclosed herein, particularly example 11 , further comprising a light-emitting module configured to be attached to the multichannel printhead and configured for photo-crosslinking of bioink.
  • Example 15 A temperature control module configured for use with a handheld bioprinter, the temperature control module comprising: a central channel configured to receive one or more bioink cartridges; an interior wall defining the central channel; an exterior wall; and one or more chambers disposed between the interior wall and the exterior wall; wherein each of the one or more chambers has a phase-change material disposed therein.
  • Example 16 The temperature control module of any example disclosed herein, particularly example 15, wherein the phase-change material comprises a first phase-change material that is selected based on a viscosity of a first bioink contained within a first one of the one or more bioink cartridges, the first phase-change material disposed within a first chamber of the one or more chambers that is proximate to the first bioink cartridge when the one or more bioink cartridges are received within the central channel.
  • Example 17 The temperature control module of any example disclosed herein, particularly example 16, wherein the phase-change material comprises a second phase-change material that is selected based on a viscosity of second bioink contained within a second one of the one or more bioink cartridges, the second phase-change material disposed within a second chamber of the one or more chambers that is proximate to the second bioink cartridge when the one or more bioink cartridges are received within the central channel, and wherein the first phase-change material is different from the second phase-change material.
  • the phase-change material comprises a second phase-change material that is selected based on a viscosity of second bioink contained within a second one of the one or more bioink cartridges, the second phase-change material disposed within a second chamber of the one or more chambers that is proximate to the second bioink cartridge when the one or more bioink cartridges are received within the central channel, and wherein the first phase-change material is different from the second phase-change material.
  • Example 18 A printhead configured for use with a bioprinter, the printhead comprising: a base portion comprising an interface configured to enable coupling of three or more bioink cartridges thereto; a tip portion having an opening configured for output of bioink; and three or more channels that extend through the base and the tip portion and configured to enable flow of bioink from the three or more bioink cartridges through the base and the tip portion to the opening.
  • Example 19 The printhead of any example disclosed herein, particularly example 18, wherein the three or more channels have a multi-compartment configuration.
  • Example 20 The printhead of any example disclosed herein, particularly example 18, wherein the three or more channels have a multi-axial configuration.
  • Example 21 The printhead of any example disclosed herein, particularly example 18, wherein the three or more channels have a multi-core configuration.
  • a bioprinting system comprising: a bioprinter comprising a printhead configured to have three or more bioink cartridges fluidly coupled thereto: a syringe pump subsystem comprising three or more syringe pumps, each of the three or more syringe pumps configured for fluid communication with one of the three or more bioink cartridges and configured to control flow of bioink from the respective bioink cartridge; and a positioning subsystem having the bioprinter coupled thereto; and a computerized controller configured to control movement of the bioprinter via the positioning system.
  • Example 23 A printhead configured for use with a bioprinter, the printhead comprising: a base portion comprising an interface configured to enable coupling of two or more bioink cartridges thereto; a tip portion having an opening configured for output of bioink; two or more channels that extend from the interface through the base; and a mixing channel that extends through the tip portion; wherein the mixing channel is configured such that bioink flowing therethrough has a non-linear flow pathway to cause mixing of bioink during flow through the tip portion to the opening.
  • Example 24 The printhead of any example disclosed herein, particularly example 23, wherein the mixing channel comprises a corkscrew member including a plurality of openings, the plurality of openings defining the non-linear flow pathway.
  • Example 25 An assembly for adapting a 3D printer to a 3D bioprinter system, the assembly comprising: bioprinter unit comprising a printhead configured to have one or more bioink cartridges fluidly coupled thereto; a syringe pump subsystem comprising one or more syringe pumps, each of the one or more syringe pumps configured for fluid communication with one of the one or more bioink cartridges and configured to control flow of bioink from the respective bioink cartridge, the syringe pump subsystem configured to be mounted to a frame of the 3D printer; and a carriage for coupling the bioprinter unit to a robotic positioning assembly of the 3D printer
  • Example 26 The assembly of any example disclosed herein, particularly example 25, further comprising a graphical user interface (GUI) configured to be brought into data communication with the 3D printer.
  • GUI graphical user interface
  • Example 27 The assembly of any example disclosed herein, particularly example 25, further comprising a computer readable storage media having one or more computer-readable instructions stored thereon for controlling operation of the 3D bioprinter system.
  • Example 28 The assembly of any example disclosed herein, particularly example 25, wherein the carriage comprises a base portion including a channel configured to be mounted to an x-axis rail of a positioning assembly of the 3D printer.
  • Example 29 The assembly of any example disclosed herein, particularly example 25, wherein the carriage comprises a magnetic plate member that is magnetically couplable to the base portion, wherein the plate member include an adaptor for receiving a portion of the bioprinter unit for coupling of the bioprinter unit thereto.
  • Example 30 A bioprinter assembly comprising: a bioprinter unit comprising a printhead configured to have one or more bioink cartridges fluidly coupled thereto; and a syringe pump subsystem comprising one or more syringe pumps, each of the one or more syringe pumps configured for fluid communication with one of the one or more bioink cartridges and configured to control flow of bioink from the respective bioink cartridge.
  • Example 31 The bioprinter assembly of any example disclosed herein, particularly example 30, further comprising a carriage for coupling the bioprinter unit to a robotic positioning subsystem.
  • Example 32 The bioprinter assembly of any example disclosed herein, particularly example 31 , wherein the carriage comprises a base portion having at least one channel for receiving an x-axis rail of the robotic positioning subsystem, and a magnetic plate member comprising an adapter configured to receive a portion of the bioprinter unit for coupling of the bioprinter unit thereto, and wherein the releasable plate member is magnetically couplable to the base.
  • Example 33 The bioprinter assembly of any example disclosed herein, particularly example 30, wherein the printhead comprises one or more female Luer lock adapters each configured to have one of the one or more bioink cartridges coupled thereto.
  • Example 34 The bioprinter assembly of any example disclosed herein, particularly example 33, further comprising the one or more bioink cartridges, wherein each of the one or more bioink cartridges comprises a male Luer lock adapter configured to mate with one of the one or more female Luer lock adapters.
  • Example 35 The bioprinter assembly of any example disclosed herein, particularly example 34, wherein each of the one or more bioink cartridges comprises a hydraulic cartridge having a piston and a hydraulic fluid disposed therein.
  • Example 36 The bioprinter assembly of any example disclosed herein, particularly example 30, further comprising a temperature control module comprising a channel configured to receive the three or more bioink cartridges therein.
  • Example 37 The bioprinter assembly of any example disclosed herein, particularly example 36, wherein the temperature control module further comprises an interior wall defining the channel, an exterior wall, a chamber defined between the interior wall and the exterior wall, and a phase change material disposed within the chamber.
  • Example 38 The bioprinter assembly of any example disclosed herein, particularly example 30, further comprising a light-emitting device configured to be attached to the printhead, the light-emitting device comprising one or a plurality of light sources configured to emit light at a selected wavelength for photo-crosslinking of bioink.
  • Example 39 The bioprinter assembly of any example disclosed herein, particularly example 30, wherein the printhead is configured to have three or more bioink cartridges fluidly coupled thereto.

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Abstract

Des bio-imprimantes, des systèmes de bio-imprimante et des ensembles pour adapter une imprimante 3D à une bio-imprimante 3D, ainsi qu'une application et des procédés associés. Les bio-imprimantes peuvent comprendre une tête d'impression multi-matériau conçue pour être couplée à deux cartouches ou plus contenant de la bio-encre ou d'autres matériaux fluides. Chacune de la cartouche peut être couplée à une pompe à seringue hydraulique dans un réseau de pompes à seringue. Dans certains exemples, les bio-imprimantes peuvent comprendre un module de commande de température ajusté autour des cartouches et/ou un module de photodurcissement ajusté sur la tête d'impression. Dans certains exemples, un ensemble pour adapter une imprimante 3D à une bio-imprimante peut comprendre les composants de bio-imprimante précédents et le réseau de pompes hydrauliques, ainsi qu'un chariot pour coupler la bio-imprimante à un ensemble bras robotique de l'imprimante 3D, une GUI et/ou un support de stockage comprenant des instructions pour faire fonctionner la bio-imprimante 3D adaptée.
PCT/IB2023/059630 2022-09-27 2023-09-27 Systèmes et appareil de bio-imprimante et procédés d'utilisation Ceased WO2024069486A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160288414A1 (en) * 2013-11-04 2016-10-06 University Of Iowa Research Foundation Bioprinter and methods of using same
US20180243478A1 (en) * 2015-09-04 2018-08-30 The General Hospital Corporation Three dimensional microtissue bioprinter
US20200023172A1 (en) * 2018-07-19 2020-01-23 Sunnybrook Research Institute Devices and methods for wound-conformal guidance of bioprinter printhead
US20200040291A1 (en) * 2016-10-07 2020-02-06 The Governing Council Of The University Of Toronto Tissue printer

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160288414A1 (en) * 2013-11-04 2016-10-06 University Of Iowa Research Foundation Bioprinter and methods of using same
US20180243478A1 (en) * 2015-09-04 2018-08-30 The General Hospital Corporation Three dimensional microtissue bioprinter
US20200040291A1 (en) * 2016-10-07 2020-02-06 The Governing Council Of The University Of Toronto Tissue printer
US20200023172A1 (en) * 2018-07-19 2020-01-23 Sunnybrook Research Institute Devices and methods for wound-conformal guidance of bioprinter printhead

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