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WO2024168145A2 - Fils à détection multiple pouvant être portés pour une surveillance continue sans fil de lactate de sueur - Google Patents

Fils à détection multiple pouvant être portés pour une surveillance continue sans fil de lactate de sueur Download PDF

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Publication number
WO2024168145A2
WO2024168145A2 PCT/US2024/014995 US2024014995W WO2024168145A2 WO 2024168145 A2 WO2024168145 A2 WO 2024168145A2 US 2024014995 W US2024014995 W US 2024014995W WO 2024168145 A2 WO2024168145 A2 WO 2024168145A2
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Prior art keywords
fiber
lactate
electrochemical
enzymatic
rgo
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WO2024168145A3 (fr
Inventor
Fiorenzo G. Omenetto
Bradley Stauffer NAPIER
Giuseppina MATZEU
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Tufts University
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Tufts University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/14532—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue for measuring glucose, e.g. by tissue impedance measurement
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/14539—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue for measuring pH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/14546—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue for measuring analytes not otherwise provided for, e.g. ions, cytochromes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • C12Q1/005—Enzyme electrodes involving specific analytes or enzymes

Definitions

  • Fibers have recently been produced from other 2D materials, such as MXenes, with excellent electrical and electrochemical properties.
  • rGO fibers are the most economically viable option as they can be produced using graphite flake - an inexpensive bulk material either from natural sources, or synthesized at low cost. These polymer-free fibers are self-assembled from monolayer graphene oxide flakes, then reduced, forming tough, conductive fibers held together by Van der Waals forces. While short segments of rGO fibers have been used as temperature sensors, gas sensors, and benchtop hydrogen peroxide sensors, the inventors have surprisingly determined that rGO fibers can be utilized in wearable enzymatic sensors.
  • the present disclosure provides an electrochemical sensing material including an enzymatic fiber and at least one of a reference fiber and a counter electrode fiber.
  • the enzymatic fiber is configured to sense a target chemical.
  • the enzymatic fiber and the at least one of the reference fiber and the counter electrode fiber are braided to form a yarn configured for weaving into a patch sensor.
  • the electrochemical sensing material also includes a circuit electrically connected to the yam.
  • the circuit is programmed to record and transmit a sensing signal.
  • the sensing signal includes one or more physiological parameters sensed by the yam.
  • the present disclosure provides a method of making an enzymatic fiber, the method includes: depositing a plurality of metallic nanoparticles onto a conductive fiber, thereby forming a first stage fiber; coating the first stage fiber with a permselective membrane as an interference rejection layer, thereby forming a second stage fiber; coating the second stage fiber with a biopolymer matrix comprising at least one enzyme to form a third stage fiber, wherein the biopolymer matrix is optionally pre-crosslinked prior to coating the second stage fiber; and coating the third stage fiber with a diffusion limiting layer, thereby forming an enzymatic fiber.
  • the present disclosure provides an enzymatic fiber including a plurality of metallic nanoparticles deposited on a reduced graphene oxide (rGO) fiber and a biopolymer membrane including lactate oxidase disposed about the rGO fiber.
  • rGO reduced graphene oxide
  • the present disclosure provides an electrochemical fiber.
  • the electrochemical fiber includes a conductive fiber and a biopolymer membrane coating disposed about the conductive fiber.
  • the biopolymer membrane includes lactate oxidase, chitosan, and silk fibroin.
  • the biopolymer matrix is optionally pre-crosslinked prior to application to the conductive fiber.
  • the present disclosure provides a lactate sensor.
  • the lactate sensor includes an electrochemical yarn including an enzymatic fiber, a reference electrode, and a counter electrode.
  • the enzymatic fiber includes a conductive fiber and a biopolymer membrane.
  • the biopolymer membrane includes lactate oxidase, chitosan, and silk fibroin.
  • the biopolymer matrix is optionally pre-crosslinked prior to application to the conductive fiber.
  • the present disclosure provides a lactate sensor.
  • the lactate sensor includes an electrochemical fiber, a reference electrode fiber, a counter electrode fiber, and a pH ion- selected electrode (ISE) fiber.
  • the electrochemical fiber includes a platinum (Pt) nanoparticle- functionalized (NP-functionalized) reduced graphene oxide (rGO) fiber, an interference rejection layer, a biopolymer member, and a diffusion limiting layer.
  • the biopolymer membrane includes lactate oxidase (LOx), chitosan, and silk fibroin. The biopolymer membrane is optionally precrosslinked prior to application to the Pt NP- functionalized rGO fiber.
  • Fig. 1 depicts an overview of the braided rGO-fiber sensor platform.
  • the multi-sensing lactate braid consists of 4 fibers - 3 fibers for the lactate sensor, one for pH calibration.
  • the lactate sensor fibers are built as follows: one rGO fiber with spray-coated reference membrane, a platinized rGO fiber as a counter electrode, and a platinized rGO fiber coated with the enzymatic membrane for lactate detection.
  • the pH sensor is a spray coated polymeric ISE membrane on a ferrocene functionalized rGO transducer fiber.
  • Fig, 3 depicts the following:
  • A Diagram depicting elements of lactate sensor fiber.
  • B Chronoamperogram of lactate sensor.
  • C Calibration curve extracted from chronoamperograms (3 replicates)
  • D Interference test of lactate sensor in lOmM phosphate buffer (pH 7.4). 1 : 5mM lactate, 2: lOmM Na + , 3: ImM Ca 2+ , 4: ImM Mg 2+ , 5: ImM glucose, 6: lOOpM ascorbic acid.
  • E Temperature response of lactate sensor in lOmM phosphate buffer and lOmM lactate.
  • F pH response of lactate sensor in lOmM phosphate/citrate buffer with lOmM lactate (3 replicates). All lactate sensors had coated lengths of 15mm.
  • Fig. 4 depicts the following: (A) On body testing setup including readout electronics, sensor braid and encapsulation patch. (B) Screen capture of iOS application with real-time readout of pH and lactate sensor values. (C) Exploded diagram of multi-layer encapsulation patch for on body testing. (D) Verification of mobile readout electronics with benchtop testing showing simultaneous outputs from pH and lactate sensors, and an external reference electrode (Ag/AgCl). NaOH ( ⁇ 0.4 pH change) and lactate (ImM) were added showing raw output (in mV) with no observable crosstalk aside from stirring induced noise. (E) On body testing data from participant 1 collected during stationary biking. Simultaneous readout of pH and lactate data from the sensor patch correlated with a skin pH probe (Hanna) and L-lactate assay kit (recorded downstream from collected sweat at multiple time points).
  • A On body testing setup including readout electronics, sensor braid and encapsulation patch.
  • B Screen capture of iOS application with real-time read
  • Fig. 5 depicts a system for weavable, multi-sensing yarns for continuous, wireless sweat lactate monitoring.
  • a graphene-based yarn is developed to be used as a weavable lactate sensor.
  • the yarn incorporates a counter electrode, reference electrode, and a pH sensor.
  • a custom wireless module and an associated app are used to demonstrate real-time, continued measurement over 45 minutes of lactate in sweat during exercise.
  • Fig. 6 depicts the following: (A) Stability of PtNP-rGO fiber in ImM Hifh and lOOmM NaCl. (B) Stability of lactate sensor in 20mM lactate with lOmM phosphate buffer.
  • Fig. 7 depicts the following: Electrochemical impedance spectroscopy (EIS) spectra (200kHz - lOOmHz, lOmV amplitude) of rGO and PtNP-rGO in 5mM ferri/ferrocyanide and lOOmM KC1.
  • EIS Electrochemical impedance spectroscopy
  • Fig. 8 depicts SEM images of PtNP-Silk/Chit/LOx-PU fiber with (A) 1 % PU spray coat, (B) 0.25% PU spray coat (Left: 20 pm scale bar, Middle: 2 pm scale bar). Lower concentration of PU gives more uniform coating of PU leading to higher linear range with similar current outputs (plots on the right for both A and B).
  • Fig. 9 depicts Fiber-based pH sensor output measured in varied pH buffer solutions (ImM phosphate-citrate-borate buffer). Measurements were paused for 1 minute in between buffer changes. Inset: Extracted calibration curve with near Nerstian slope (55.8 mV pH 1 ).
  • Fig. 10 depicts readout electronics: Front and back of custom readout PCB utilizing a Nordic NRF52832 BLE SOC, high input impedance voltage buffer, and LMP91000 potentiostat with supporting circuitry and CR1215 coin cell battery.
  • Fig. 11 depicts Participant 2 - Simultaneous pH and lactate data with correlated pH (Hanna skin probe) and lactate measurements (L-lactate assay on collected sweat, multiple time points) during stationary biking exercise. Slow hydration of the pH sensor led to a delayed stabilization of the sensor.
  • Fig. 12 depicts the results of a wicking test of encapsulation patch for on body testing. 50 pL of dye was pipetted on the inlet opening. Pictures were taken at 10, 20, 40, and 60 seconds.
  • Fig. 13 is a schematic that summarizes the PtNP-rGO fiber preparation.
  • the term “a” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and/or”; (iii) the terms “comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; and (iv) the terms “about” and “approximately” are used as equivalents and may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (v) where ranges are provided, endpoints are included.
  • composition as used herein, may be used to refer to a discrete physical entity that comprises one or more specified components.
  • a composition may be of any form - e.g., gas, gel, liquid, solid, etc.
  • composition may refer to a combination of two or more entities for use in a single embodiment or as part of the same article.
  • the combination of entities result in physical admixture, that is, combination as separate co-entities of each of the components of the composition is possible; however many practitioners in the field may find it advantageous to prepare a composition that is an admixture of two or more of the ingredients in a pharmaceutically acceptable carrier, diluent, or excipient, making it possible to administer the component ingredients of the combination at the same time.
  • the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest.
  • One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result.
  • the term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.
  • silk fibroin refers to silk fibroin protein whether produced by silkworm, spider, or other insect, or otherwise generated (Lucas et al., Adv. Protein Chem., 13: 107-242 (1958)). Any type of silk fibroin can be used in different embodiments described herein.
  • Silk fibroin produced by silkworms, such as Bombyx mori is the most common and represents an earth-friendly, renewable resource.
  • silk fibroin used in a silk film may be attained by extracting sericin from the cocoons of B. mori.
  • Organic silkworm cocoons are also commercially available.
  • silks there are many different silks, however, including spider silk (e.g., obtained from Nephila clavipes), transgenic silks, genetically engineered silks, such as silks from bacteria, yeast, mammalian cells, transgenic animals, or transgenic plants, and variants thereof, that can be used. See, e.g., WO 97/08315 and U.S. Pat. No. 5,245,012, each of which is incorporated herein by reference in their entireties.
  • spider silk e.g., obtained from Nephila clavipes
  • transgenic silks e.g., obtained from Nephila clavipes
  • genetically engineered silks such as silks from bacteria, yeast, mammalian cells, transgenic animals, or transgenic plants, and variants thereof, that can be used. See, e.g., WO 97/08315 and U.S. Pat. No. 5,245,012, each of which is incorporated herein by reference in their entireties.
  • a “fiber” refers to an article having a significant length dimension in comparison to its lateral size dimensions, with a minimum length (which is required for weaving) and maximum lateral size dimensions, with diameters of between 5 pm and 100 pm or between 5 pm and 25 pm.
  • Textile integrated sensors based on conductive, electrochemically active microfibers can enable inexpensive, nearly invisible distributed sensing of sweat in clothing.
  • Reduced graphene oxide (rGO) fibers are mechanically robust, conductive, and can be functionalized to form sensors with properties comparable to planar fabricated sensors, given their ability to work both as electrical interconnections and as a base electrode.
  • rGO-based fibers are a promising material platform for embedded textile biosensor electrodes due to the inherent electrical, mechanical, and electrochemical properties of graphene, ease of fabrication, tunability of the fibers, and inexpensive material cost. Applications for these fibers range from low-cost disposable chemical sensors to wearable energy storage to lightweight interconnects.
  • nanomaterial functionalization of these fibers enables simultaneous pH and lactate sensing in a convenient textile-ready format.
  • Solution-based processing allows for facile coating of large lengths of fiber with metallic nanoparticles, such as platinum nanoparticles, with stable performance as a mediator for hydrogen peroxide. Further coatings with Nafion, biopolymer embedded lactate oxidase, and a spray-coated diffusion layer of polyurethane enables sensing of lactate in sweat with extended linear range. Realtime sweat sensing from a pH/lactate fiber bundle with high correlation to external measurements demonstrates the potential of these fibers in a wearable electrochemical sensing configuration. Also disclosed herein is a platform of textile-integrated “electrochemical yams” of broad utility and use enabled by functionalization with various ID and 2D nanomaterials and coatings with active membranes.
  • an electrochemical yarn based on modified dry-spun rGO fibers contains a reference electrode, a counter electrode, an enzyme-responsive lactate oxidase functionalized fiber (e.g., a lactate-responsive fiber functionalized with lactate oxidase), and, optionally, a pH-sensing fiber for calibration in a single, robust, weavable format.
  • This “electrochemical yarn” was integrated into a demonstrator wearable textile-patch capable of continuous data collection and wireless data transmitted to an ad-hoc app. Surprisingly, the yarns perform comparably to traditional probes in a format of broad utility for standalone or integrated monitoring of physiological parameters.
  • pure rGO fibers are an excellent base material as they can be tailored to suit the application’s needs with a multitude of functionalization possibilities.
  • functional groups in graphene e.g. epoxy, hydroxyl, carboxyl
  • the surface can be tailored to the application, increasing capacitance by adding redox groups (e.g. ferrocene), altering mechanical or electrical properties, or adding specific functional groups to enhance post-process modifications.
  • the fibers can also be doped with various nanomaterials from 0D to 2D such as MXenes, cellulose nanocrystals, metallic nanoparticles, increasing strength or altering electrical properties.
  • the reduced fibers can also be functionalized in the same way as rGO, with either electro- or electroless deposition of nanomaterials and/or conducting polymers.
  • Dip coating and spray coating enables chemically selective electrodes, reference electrodes, and insulating coatings. Combining various functionalization approaches enables numerous textile integrated applications in biochemical and bioelectrical sensing as well as photovoltaics and supercapacitors.
  • rGO has been used as the base electrode or with additives such as metal catalyst nanostructures, redox moieties and Prussian Blue (PB) among others for improved performance. Functionalization with these materials allows for either indirect measurement of enzyme activity through hydrogen peroxide (e.g. mediated by platinum, PB) or direct measurement through redox mediators (e.g. tetrathiafulvalene) at lower operating potentials.
  • additives such as metal catalyst nanostructures, redox moieties and Prussian Blue (PB) among others for improved performance.
  • PB Prussian Blue
  • rGO yarns for wearable sweat sensing are disclosed herein.
  • bundles of rGO fibers can enable the fabrication of electrochemical yarns useful for both textile integrated sensing applications (sweat, gas, viral/microbial) as well as a potential low waste disposable sensor alternative to screen printed carbon electrodes. Screen-printing generates a lot of non-recyclable waste and uses harsh chemicals, making it less sustainable than the proposed fibers.
  • these small form factor ( ⁇ 100 pm diameter) multiplexed sensor yarns are constituted by bundling a reference, counter electrode, and pH sensing fibers in a single convenient format.
  • Sweat contains a variety of salts, metabolic products, and other biomarkers indicative either of overall health or current state (fatigue, stress). Often measured is L-lactate, the metabolic product produced at increasing rates with muscle fatigue. This is typically measured with colorimetric or electrochemical sensors using lactate oxidase (LOx). Although specific to L-lactate, these sensors have various redox and reactive species (e.g., ascorbic acid, uric acid) whose interference can be avoided using selectively permeable membranes underneath the enzymatic layer (e.g. p- Phenylenediamine (PPD), Nafion). Other parameters, such as temperature and pH, can be simultaneously measured to allow for error correction in current output.
  • LOx lactate oxidase
  • the electrochemical sensing material includes an enzymatic fiber configured to sense a target chemical, such as lactate.
  • the electrochemical sensing material also includes at least one of a reference fiber and a counter electrode fiber.
  • the enzymatic fiber and at least one of the reference fiber and counter electrode fiber are braided, or otherwise disposed adjacently, to form a yam. In some embodiments, the yam is not spun.
  • the yam may be configured for weaving into a patch sensor.
  • the electrochemical sensing material includes a circuit electrically connected to the yarn.
  • the circuit is programmed to record and transmit a sensing signal, such as including one or more physiological parameters sensed by the yarn.
  • the circuit may also be programmed to record and transmit a sensing signal including one or more environmental, biochemical viral/microbial, gaseous, or bioelectrical parameters sensed by the yarn.
  • the electrochemical sensing material may also include a processor configured to receive the sensing signal and output one or more parameters (e.g., physiological, biochemical, bioelectrical, viral/microbial, and/or environmental parameters) sensed by the yarn.
  • the circuit in electrical communication with the yam may transmit the sensing signal wirelessly to the processor.
  • the enzymatic fiber and at least one of the reference fiber and counter electrode fiber may be or may have cores that are reduced graphene oxide (rGO) fibers.
  • the enzymatic fiber may be coated in a biopolymer matrix including lactate oxidase (LOx), chitosan, and silk fibroin, such as to facilitate sensing lactate as a target chemical.
  • the enzymatic fiber includes a selectively permeable membrane positioned between the fiber and biopolymer matrix configured to prevent redox and reactive species interference.
  • the enzymatic fiber may include a layer of polyurethane (PU) at least partially coating the biopolymer matrix.
  • the enzymatic fiber and counter electrode fiber further include platinum nanoparticles (Pt NPs).
  • the enzymatic fiber is doped with at least one nanomaterial.
  • the reference fiber is coated in a layer of heterogeneous PVC- Ag/AgCl-KCl.
  • the yarn further includes a pH sensing fiber that may be a ferrocene modified rGO fiber coated in a polymeric pH ion selective membrane.
  • the yarn may also include a temperature sensing fiber. Sensing signals from the pH and/or temperature sensing fibers may be used for error correction in current output from the electrochemical sensing material.
  • the enzymatic fiber includes one or more additives, such as metal catalyst nanostructures, redox moieties, or Prussian Blue (PB).
  • additives such as metal catalyst nanostructures, redox moieties, or Prussian Blue (PB).
  • an electrochemical sensing material including depositing a platinum nanoparticle on a fiber.
  • the platinum nanoparticle-coated fiber is then coated with a permselective membrane, such as Nafion, as an interference rejection layer.
  • the fiber is further coated with a biopolymer matrix comprising at least one enzyme to form an enzymatic fiber.
  • the biopolymer matrix may include lactate oxidase (LOx), chitosan, and silk fibroin.
  • the biopolymer matrix is optionally pre-crosslinked, as disclosed elsewhere herein.
  • An electrochemical yarn is formed by intertwining or braiding the enzymatic fiber with a counter electrode fiber, which may be coated with the platinum nanoparticle, and a reference electrode fiber.
  • the electrochemical yarn can be interfaced to a socket connector to form a sensing yarn.
  • the sensing yam may be electrically connected to a circuit programmed to record and transmit a sensing signal including one or more physiological, environmental, biochemical, viral, microbial, or bioelectrical parameters sensed by the yarn.
  • the sensing yarn may be integrated into a wearable patch.
  • a first stage fiber may be formed by depositing a plurality of metallic nanoparticles onto a fiber, such as a conductive fiber.
  • the plurality of metallic nanoparticles may include a metal, such as one selected from the group consisting of platinum, silver, gold, palladium, zinc, iron, copper, molybdenum, cobalt, and combinations thereof.
  • a diameter of the first stage fiber may be between 20 pm and 50 pm.
  • the diameter of the first stage fiber may be at least 20 pm, at least 30 pm, or at least 50 pm.
  • the diameter of the first stage fiber may be at most 50 pm, at most 35 pm, or at most 30 pm.
  • a second stage fiber may be formed by coating the first stage fiber with a permselective membrane (e.g. Nafion) as an interference rejection layer.
  • a diameter of the second stage fiber may be between 25 pm and 60 pm.
  • the diameter of the second stage fiber may be at least 25 pm, at least 40 pm, or at least 55 pm.
  • the diameter of the second stage fiber may be at most 60 pm, at most 45 pm, or at most 35 pm.
  • a third stage fiber may be formed by coating the second stage fiber with a biopolymer matrix comprising at least one enzyme (e.g., lactate oxidase).
  • the biopolymer matrix may include lactate oxidase (LOx), chitosan, and silk fibroin.
  • the biopolymer matrix is optionally pre-crosslinked prior to coating the second stage fiber, such as by glutaraldehyde.
  • a thickness of the biopolymer matrix may be between 5 pm and 10 pm.
  • a thickness of the biopolymer matrix may be at least 5 pm or at least 7 pm.
  • a thickness of the biopolymer matrix may be at most 10 pm or at most 7 pm.
  • the diameter of the third stage fiber may be between 30 pm and 70 pm.
  • the diameter of the third stage fiber may be at least 30 pm, at least 40 pm, or at least 60 pm.
  • the diameter of the third stage fiber may be at most 70 pm, at most 55 pm, or at most 35 pm.
  • the enzymatic fiber may be formed by coating the third stage fiber with a diffusion limiting layer, such as polyurethane.
  • the diameter of the enzymatic fiber may be between 35 pm and 80 pm.
  • the diameter of the enzymatic fiber may be at least 35 pm, at least 60 pm, or at least 70 pm.
  • the diameter of the enzymatic fiber may be at most 80 pm, at most 65 pm, or at most 45 pm.
  • an electrochemical yarn may be formed by intertwining or braiding the enzymatic fiber with a counter electrode fiber and a reference electrode fiber.
  • the counter electrode fiber may be coated with a second plurality of metallic nanoparticles, which may include a metal selected from the group consisting of platinum, silver, gold, palladium, zinc, iron, molybdenum, cobalt, copper, and combinations thereof.
  • the first and second plurality of metallic nanoparticles may be the same metal or combination of metals, or in other embodiments, may be a different metal or combination of metals.
  • the diameter of the electrochemical yarn may be between 95 pm and 230 pm.
  • the diameter of the electrochemical yarn may be at least 95 pm, at least 100 pm, at least 120 pm, or at least 150 pm.
  • the diameter of the electrochemical yarn may be at most 230 pm, at most 200 pm, at most 180 pm, or at most 170 pm.
  • the enzymatic fiber may also be intertwined with a pH ion-selective electrode (ISE) fiber and/or a temperature-sensing fiber in addition to the counter electrode fiber and the reference electrode fiber to form the electrochemical yam.
  • ISE pH ion-selective electrode
  • the electrochemical yarn may interface with a connector (e.g., socket connector) to form a sensing yarn which may be integrated into a wearable patch.
  • the sensing yarn may be electrically connected to a circuit, wherein the circuit is programmed to record and transmit a sensing signal including one or more physiological, environmental, biochemical, viral, microbial, or bioelectrical parameters sensed by the yam.
  • a platinized reduced graphene oxide (rGO) fiber including a plurality of platinum nanoparticles deposited on an rGO fiber by electro-deposition or by an electroless, solutionbased deposition.
  • the platinized rGO fiber may include a coating to enable at least one of a chemically selective electrode, a reference electrode, or an insulating coating.
  • the coating may be a biopolymer membrane including lactate oxidase, chitosan, and silk fibroin which is optionally cross-linked, such as by glutaraldehyde, which has a thickness of between 5 pm and 10 pm.
  • the thickness of the biopolymer membrane may be at least 5 pm, at least 7 pm, or at least 10 pm. In some embodiments, the thickness of the biopolymer membrane may be at most 10 pm, at most 7 pm, or at most 5 pm.
  • the diameter of the platinized rGO fiber may be between 20 pm and 50 pm. The diameter of the platinized rGO fiber may be at least 20 pm, at least 30 pm, or at least 45 pm. The diameter of the platinized rGO fiber may be at most 50 pm, at most 35 pm, or at most 30 pm.
  • an enzymatic fiber including a plurality of metallic nanoparticles deposited on a reduced graphene oxide (rGO) fiber, and a biopolymer membrane comprising lactate oxidase disposed about the rGO fiber to form the enzymatic fiber.
  • the plurality of metallic nanoparticles may be deposited by electro-deposition or by an electroless, solution-based deposition.
  • the plurality of metallic nanoparticles may include a metal selected from the group consisting of platinum, silver, gold, palladium, zinc, iron, copper, molybdenum, cobalt, and combinations thereof.
  • the biopolymer membrane may be pre-crosslinked, such as by glutaraldehyde.
  • the thickness of the biopolymer membrane may be between 5 pm and 10 pm.
  • the thickness of the biopolymer membrane may be at least 5 pm or at least 7 pm, or higher.
  • the thickness of the biopolymer membrane may be at most 10 pm, at most 7 pm, or lower.
  • the diameter of the enzymatic fiber may be between 20 pm and 50 pm.
  • the diameter of the enzymatic fiber may be at least 20 pm, at least 30 pm, or at least 45 pm.
  • the diameter of the enzymatic fiber may be at most 50 pm, at most 35 pm, or at most 30 pm.
  • an electrochemical fiber including a conductive fiber, and a biopolymer membrane coating disposed about the conductive fiber.
  • the diameter of the conductive fiber may be between 20 pm and 50 pm.
  • the diameter of the conductive fiber may be at least 20 pm, at least 30 pm, or at least 45 pm.
  • the diameter of the conductive fiber may be at most 50 pm, at most 35 pm, or at most 30 pm.
  • the biopolymer membrane includes lactate oxidase, chitosan, and silk fibroin.
  • the biopolymer matrix is optionally pre-crosslinked prior to application to the conductive fiber, such as with glutaraldehyde.
  • the thickness of the biopolymer matrix may be between 5 pm and 10 pm.
  • the thickness of the biopolymer matrix may at least 5 pm, at least 7 pm, or at least 9 pm.
  • the thickness of the biopolymer matrix may be at most 10 pm, at most 7 pm, or at most 6 pm.
  • the diameter of the conductive fiber and the biopolymer membrane may be between 25 pm and 60 pm.
  • the diameter of the conductive fiber and the biopolymer membrane may be at least 25 pm, at least 40 pm, or at least 55 pm.
  • the diameter of the conductive fiber and the biopolymer membrane may be at most 60 pm, at most 45 pm, or at most 35 pm.
  • the conductive fiber may be an rGO fiber functionalized with metallic nanoparticles, including a metal selected from the group consisting of platinum, silver, gold, palladium, zinc, iron, molybdenum, cobalt, copper, and combinations thereof.
  • the metallic nanoparticles may be deposited by a solution-based process or by electro-deposition.
  • the electrochemical fiber may further include a selectively permeable membrane disposed between the conductive fiber and the biopolymer matrix.
  • the thickness of the selectively permeable membrane may be between 5 pm and 10 pm.
  • the thickness of the selectively permeable membrane may be at least 5 pm, at least 7 pm, or at least 9 pm.
  • the thickness of the selectively permeable membrane may be at most 10 pm, at most 7 pm, or at most 6 pm.
  • the diameter of the conductive fiber, the biopolymer membrane, and the selectively permeable membrane may be between 30 pm and 70 pm.
  • the diameter of the conductive fiber, the biopolymer membrane, and the selectively permeable membrane may be at least 30 pm, at least 40 pm, or at least 60 pm.
  • the diameter of the conductive fiber, the biopolymer membrane, and the selectively permeable membrane may be at most 70 pm, at most 55 pm, or at most 35 pm.
  • the electrochemical fiber may further include a diffusion limiting layer disposed on the biopolymer membrane, such as polyurethane.
  • the thickness of the diffusion limiting layer may be between 5 pm and 10 pm.
  • the thickness of the diffusion limiting layer may be at least 5 pm, at least 7 pm, or at least 9 pm.
  • the thickness of the diffusion limiting layer may be at most 10 pm, at most 7 pm, or at most 6 pm.
  • the diameter of the conductive fiber, the biopolymer membrane, the selectively permeable membrane, and the diffusion limiting layer may be between 35 pm and 80 pm.
  • the diameter of the conductive fiber, the biopolymer membrane, the selectively permeable membrane, and the diffusion limiting layer may be at least 35 pm, at least 60 pm, or at least 70 pm.
  • the diameter of the conductive fiber, the biopolymer membrane, the selectively permeable membrane, and the diffusion limiting layer may be at most 80 pm, at most 65 pm, or at most 45 pm.
  • the electrochemical fiber may be configured to be combined with a reference electrode fiber and a counter electrode fiber to form a three-fiber lactate sensor.
  • a lactate sensor including an electrochemical yam including an enzymatic fiber, a reference electrode fiber, and a counter electrode fiber.
  • the enzymatic fiber may include a conductive fiber, and a biopolymer membrane.
  • the biopolymer membrane may include lactate oxidase, chitosan, and silk fibroin, wherein the biopolymer matrix is optionally pre-crosslinked, such as with glutaraldehyde, prior to application to the conductive fiber.
  • the conductive fiber is an rGO fiber functionalized with metallic nanoparticles, which may include a metal selected from the group consisting of platinum, silver, gold, palladium, zinc, iron, copper, molybdenum, cobalt, and combinations thereof.
  • the thickness of the biopolymer membrane may be between 5 m and 10
  • the thickness of the biopolymer membrane may be at least 5 pm, at least 7 pm, or at least 9 pm.
  • the thickness of the biopolymer membrane may be at most 10 pm, at most 7 pm, or at most 6 pm.
  • the lactate sensor may further include at least one of a diffusion limiting layer or an interference rejection layer.
  • the thickness of the diffusion limiting layer or the interference rejection layer may be between 5 pm and 10 pm.
  • the thickness of the diffusion limiting layer or the interference rejection layer may be at least 5 pm, at least 7 pm, or at least 9 pm.
  • the thickness of the diffusion limiting layer or the interference rejection layer may be at most 10 pm, at most 7 pm, or at most 6 pm.
  • the lactate sensor may further include a pH ion-selective electrode (ISE) fiber.
  • the pH ISE fiber may be fabricated by spray-coating a polymeric ISE membrane on a ferrocene modified rGO (fc-rGO) fiber.
  • the diameter of the pH ISE fiber may be between 20 pm and 50 pm.
  • the diameter of the pH ISE fiber may be at least 20 pm, at least 30 pm, or at least 45 pm.
  • the diameter of the pH ISE fiber may be at most 50 pm, at most 35 pm, or at most 30 pm.
  • the lactate sensor may further include a circuit interface.
  • the diameter of the lactate sensor may be between 75 pm and 180 pm.
  • the diameter of the lactate sensor may be at least 75 pm, at least 90 pm, or at least 150 pm.
  • the diameter of the lactate sensor may be at most 180 pm, at most 100 pm, or at most 80 pm.
  • a lactate sensor including an electrochemical fiber, a reference electrode fiber (e.g., a fiber spray-coated with a layer of heterogenous PVC-Ag/AgCl-KCl), a counter electrode fiber, and a pH ion-selective electrode (ISE) fiber.
  • a reference electrode fiber e.g., a fiber spray-coated with a layer of heterogenous PVC-Ag/AgCl-KCl
  • ISE pH ion-selective electrode
  • the electrochemical fiber includes a platinum (Pt) nanoparticle-functionalized (NP-functionalized) reduced graphene oxide (rGO) fiber, an interference rejection layer, a biopolymer membrane comprising lactate oxidase (LOx), chitosan, and silk fibroin, wherein the biopolymer membrane is optionally pre-crosslinked (e.g., cross-linked with glutaraldehyde) prior to application to the Pt NP-functionalized rGO fiber, and a diffusion limiting layer (e.g., polyurethane).
  • the diameter of the Pt NP-functionalized rGO fiber, reference electrode fiber, or counter electrode fiber may be between 20 pm and 50 pm.
  • the diameter of the Pt NP-functionalized rGO fiber, reference electrode fiber, or counter electrode fiber may be at least 20 pm, at least 30 pm, or at least 45 pm.
  • the diameter of the Pt NP-functionalized rGO fiber, reference electrode fiber, or counter electrode fiber may be at most 50
  • the diameter of the lactate sensor may be between 95 m and 230 pm.
  • the diameter of the lactate sensor may be at least 95 pm, at least 120 pm, or at least 150 pm.
  • the diameter of the lactate sensor may be at most 230 pm, at most 180 pm, or at most 170 pm.
  • the thickness of the biopolymer membrane, interference rejection layer, or diffusion limiting layer may be between 5 pm and 10 pm.
  • the thickness of the biopolymer membrane, interference rejection layer, or diffusion limiting layer may be at least 5 pm, at least 7 pm, or at least 9 pm.
  • the thickness of the biopolymer membrane, interference rejection layer, or diffusion limiting layer may be at most 10 pm, at most 7 pm, or at most 6 pm.
  • the diameter of the electrochemical fiber may be between 50 pm and 75 pm.
  • the diameter of the electrochemical fiber may be at least 50 pm, at least 60 pm, or at least 70 pm.
  • the diameter of the electrochemical fiber may be at most 75 pm, at most 60 pm, or at most 55 pm.
  • the pH ISE fiber may be fabricated by spray-coating a polymeric ISE membrane on a ferrocene modified rGO (fc-rGO) fiber.
  • the lactate sensor may further include a circuit interface in electrical communication with the electrochemical yarn, reference electrode fiber, counter electrode fiber, and pH ion-selective electrode (ISE) fiber.
  • ISE pH ion-selective electrode
  • the silk fibroin may be present in an amount by weight of 1%.
  • the chitosan may be present in an amount by weight of 1%.
  • wearable lactate sensing patches including any of the lactate sensors disclosed herein.
  • the lactate sensor is integrated with a microfluidic thread structure.
  • the wearable lactate sensing patch is a microfluidic skin patch.
  • any application-appropriate amount of one or more functionalizing agents may be used.
  • the amount of an individual functionalizing agent may be between about 1 pg/ml and 1,000 pg/ml (e.g., between about 2 and 1,000, 5 and 1,000, 10 and 1,000, 10 and 500, 10 and 100 pg/ml).
  • the amount of an individual functionalizing agent may be at least 1 pg/ml (e.g., at least 5, 10, 15, 20 25, 50, 100, 200, 300 400, 500, 600, 700, 800, or 900 pg/ml ).
  • the amount of an individual functionalizing agent is at most 1,000 pg/ml (e.g., 900, 800, 700, 600, 500, 400, 300 200, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, or 5 pg/ml ).
  • the functionalizing agent may include one or more sensing agents, such as a sensing dye.
  • the sensing agents/sensing dyes are environmentally sensitive and produce a measurable response to one or more environmental factors.
  • the environmentally-sensitive agent or dye may be present in the composition in an effective amount to alter the composition from a first chemical -physical state to a second chemical -physical state in response to an environmental parameter (e.g., a change in pH, light intensity or exposure, temperature, pressure or strain, voltage, physiological parameter of a subject, and/or concentration of chemical species in the surrounding environment) or an externally applied stimulus (e.g., optical interrogation, acoustic interrogation, and/or applied heat).
  • an environmental parameter e.g., a change in pH, light intensity or exposure, temperature, pressure or strain, voltage, physiological parameter of a subject, and/or concentration of chemical species in the surrounding environment
  • an externally applied stimulus e.g., optical interrogation, acoustic interrogation, and/or applied heat.
  • the sensing dye is present to provide one optical appearance under one given set of environmental conditions and a second, different optical appearance under a different given set of environmental conditions.
  • Suitable concentrations for the sensing agents described herein can be the concentrations for the colorants and additives described elsewhere herein.
  • a person having ordinary skill in the chemical sensing arts can determine a concentration that is appropriate for use in a sensing application of the inks described herein.
  • the first and second chemical-physical state may be a physical property of the composition, such as mechanical property, a chemical property, an acoustical property, an electrical property, a magnetic property, an optical property, a thermal property, a radiological property, or an organoleptic property.
  • exemplary sensing dyes or agents include, but are not limited to, a pH sensitive agent, a thermal sensitive agent, a pressure or strain sensitive agent, a light sensitive agent, or a potentiometric agent.
  • Exemplary pH sensitive dyes or agents include, but are not limited to, cresol red, methyl violet, crystal violet, ethyl violet, malachite green, methyl green, 2-(p- dimethylaminophenylazo) pyridine, paramethyl red, metanil yellow, 4-phenylazodiphenylamine, thymol blue, metacresol purple, orange IV, 4-o-Tolylazo-o-toluindine, quinaldine red, 2,4- dinitrophenol, erythrosine disodium salt, benzopurpurine 4B, N,N-dimethyl-p-(m-tolylazo) aniline, p- dimethylaminoazobenene, 4,4'-bis(2-amino-l-naphthylazo)-2,2'-stilbenedisulfonic acid, tetrabromophenolphthalein ethyl ester, bromophenol blue, Congo red, methyl orange, ethyl orange, 4-
  • Exemplary light responsive dyes or agents include, but are not limited to, photochromic compounds or agents, such as triarylmethanes, stilbenes, azasilbenes, nitrones, fulgides, spiropyrans, napthopyrans, spiro-oxzines, quinones, derivatives and combinations thereof.
  • photochromic compounds or agents such as triarylmethanes, stilbenes, azasilbenes, nitrones, fulgides, spiropyrans, napthopyrans, spiro-oxzines, quinones, derivatives and combinations thereof.
  • Exemplary potentiometric dyes include, but are not limited to, substituted amiononaphthylehenylpridinium (ANEP) dyes, such as di-4-ANEPPS, di-8-ANEPPS, and N-(4- Sulfobutyl)-4-(6-(4-(Dibutylamino)phenyl)hexatrienyl)Pyridinium (RH237).
  • ANEP substituted amiononaphthylehenylpridinium
  • Exemplary temperature sensitive dyes or agents include, but are not limited to, thermochromic compounds or agents, such as thermochromic liquid crystals, leuco dyes, fluoran dyes, octadecylphosphonic acid.
  • Exemplary pressure or strain sensitive dyes or agents include, but are not limited to, spiropyran compounds and agents.
  • chemi-sensitive dyes or agents include, but are not limited to, antibodies such as immunoglobulin G (IgG) which may change color from blue to red in response to bacterial contamination.
  • IgG immunoglobulin G
  • the functionalizing agent comprises one or more additive, dopant, or biologically active agent suitable for a desired intended purpose.
  • the additive or dopant may be present in an amount effective to impart an optical or organoleptic property to the composition.
  • Exemplary additives or dopants that impart optical or organoleptic properties include, but are not limited to, dyes/pigments, flavorants, aroma compounds, granular or fibrous fillers.
  • the additive, dopant, or biologically active agent may be present in an amount effective to "functionalize” the composition to impart a desired mechanical property or added functionality to the composition.
  • exemplary additive, dopants, or biologically active agent that impart the desired mechanical property or added functionality include, but are not limited to: environmentally sensitive/sensing dyes; active biomolecules; conductive or metallic particles; micro and nanofibers (e.g., silk nanofibers for reinforcement, carbon nanofibers); nanotubes; inorganic particles (e.g., hydroxyapatite, tricalcium phosphate, bioglasses); drugs (e.g., antibiotics, small molecules or low molecular weight organic compounds); proteins and fragments or complexes thereof (e.g., enzymes, antigens, antibodies and antigen-binding fragments thereof);
  • DNA/RNA e.g., siRNA, miRNA, mRNA
  • cells and fractions thereof viruseses and viral particles; prokaryotic cells such as bacteria; eukaryotic cells such as mammalian cells and plant cells; fungi).
  • the additive or dopant comprises an aroma compound.
  • aroma compounds include ester aroma compounds, terpene aroma compounds, cyclic terpenes, and aromatic aroma compounds, such as, but not limited to, geranyl acetate, methyl formate, metyl acetate, methyl propionate, methyl butyrate, ethyl acetate, ethyl butyrate, isoamyl acetate, pentyl butrate, pentyl pentanoate, octyl acetate, benzyl acetate, methyl anthranilate, myrecene, geraniol, nerol, citral, cironellal, cironellol, linalool, nerolidol, limonene, camphor, menthol, carone, terpineol, alpha-lonone, thujone, eucalyptol, benzaldehyde, eugenol, cinnamaldehyde,
  • the additive or dopant comprises a colorant, such as a dye or pigment.
  • the dye or pigment imparts a color or grayscale to the composition.
  • the colorant can be different than the sensing agents and/or sensing dyes below. Any organic and/or inorganic pigments and dyes can be included in the inks.
  • Exemplary pigments suitable for use in the present disclosure include International Color Index or C.I. Pigment Black Numbers 1 , 7, 1 1 and 31 , C.I. Pigment Blue Numbers 15, 15 : 1 , 15 :2, 15 :3, 15 :4, 15 :6, 16, 27, 29, 61 and 62, C.I. Pigment Green Numbers 7, 17, 18 and 36, C.I.
  • carbon black pigment such as Regal 330, Cabot Corporation
  • quinacridone pigments Quinacridone Magenta (228-0122), available from Sun Chemical Corporation, Fort Lee, N.I.
  • diarylide yellow pigment such as AAOT Yellow (274- 1788) available from Sun
  • the classes of dyes suitable for use in present invention can be selected from acid dyes, natural dyes, direct dyes (either cationic or anionic), basic dyes, and reactive dyes.
  • the acid dyes also regarded as anionic dyes, are soluble in water and mainly insoluble in organic solvents and are selected, from yellow acid dyes, orange acid dyes, red acid dyes, violet acid dyes, blue acid dyes, green acid dyes, and black acid dyes.
  • European Patent 0745651 incorporated herein by reference, describes a number of acid dyes that are suitable for use in the present disclosure.
  • Exemplary yellow acid dyes include Acid Yellow 1 International Color Index or C.I. 10316); Acid Yellow 7 (C.I.
  • Exemplary orange acid dyes include Acid Orange 1 (C.I. 13090/1); Acid Orange 10 (C.I. 16230); Acid Orange 20 (C.I. 14603); Acid Orange 76 (C.I. 18870); Acid Orange 142; Food Orange 2 (C.I. 15980); and Orange B.
  • Exemplary red acid dyes include Acid Red 1. (C.I. 18050); Acid Red 4 (C.I. 14710); Acid Red 18 (C.I. 16255), Acid Red 26 (C.I. 16150); Acid Red 2.7 (C.I. as Acid Red 51 (C.I. 45430, available from BASF Corporation, Mt. Olive, N.J.) Acid Red 52 (C.I. 45100); Acid Red 73 (C.I. 27290); Acid Red 87 (C. I. 45380); Acid Red 94 (C.I. 45440) Acid Red 194; and Food Red 1 (C.I. 14700).
  • Exemplary violet acid dyes include Acid Violet 7 (C.I. 18055); and Acid Violet 49 (C.I. 42640).
  • Exemplary blue acid dyes include Acid Blue 1 (C.I. 42045); Acid Blue 9 (C.I. 42090); Acid Blue 22 (C.I. 42755); Acid Blue 74 (C.I. 73015); Acid Blue 93 (C.I. 42780); and Acid Blue 158A (C.I. 15050).
  • Exemplary green acid dyes include Acid Green 1 (C.I. 10028); Acid Green 3 (C.I. 42085); Acid Green 5 (C.I. 42095); Acid Green 26 (C.I. 44025); and Food Green 3 (C.I. 42053).
  • Exemplary black acid dyes include Acid Black 1 (C.I. 20470); Acid Black 194 (Basantol® X80, available from BASF Corporation, an azo/1 :2 CR-complex.
  • Exemplary direct dyes for use in the present disclosure include Direct Blue 86 (C.I. 74180); Direct Blue 199; Direct Black 168; Direct Red 253; and Direct Yellow 107/132 (C.I. Not Assigned).
  • Exemplary natural dyes for use in the present disclosure include Alkanet (C.I.
  • Exemplary reactive dyes for use in the present disclosure include Reactive Yellow 37 (monoazo dye); Reactive Black 31 (disazo dye); Reactive Blue 77 (phthalo cyanine dye) and Reactive Red 180 and Reactive Red 108 dyes. Suitable also are the colorants described in The Printing Ink Manual (5th ed., Leach et al. eds.
  • compositions provided herein can contain ETV fluorophores that are excited in the ETV range and emit light at a higher wavelength (typically 400 nm and above).
  • ETV fluorophores include but are not limited to materials from the coumarin, benzoxazole, rhodamine, napthalimide, perylene, benzanthrones, benzoxanthones or benzothia- xanthones families.
  • a UV fluorophore such as an optical brightener for instance
  • the amount of colorant, when present, generally is between 0.05% to 5% or between 0.1% and 1% based on the weight of the composition.
  • the amount of pigment/dye generally is present in an amount of from at or about 0.1 wt% to at or about 20 wt% based on the weight of the composition.
  • a non- white ink can include 15 wt% or less pigment/dye, or 10 wt% or less pigment/dye or 5 wt% pigment/dye, or 1 wt% pigment/dye based on the weight of the composition.
  • a non-white ink can include 1 wt% to 10 wt%, or 5 wt% to 15 wt%, or 10 wt% to 20 wt% pigment/dye based on the weight of the composition.
  • a non-white ink can contain an amount of dye/pigment that is 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15%, 16 wt%, 17 wt%, 18 wt%, 19 wt% or 20 wt% based on the weight of the composition.
  • the amount of white pigment generally is present in an amount of from at or about 1 wt% to at or about 60 wt% based on the weight of the composition. In some applications, greater than 60 wt% white pigment can be present.
  • Preferred white pigments include titanium dioxide (anatase and rutile), zinc oxide, lithopone (calcined coprecipitate of barium sulfate and zinc sulfide), zinc sulfide, blanc fixe and alumina hydrate and combinations thereof, although any of these can be combined with calcium carbonate.
  • a white ink can include 60 wt% or less white pigment, or 55 wt% or less white pigment, or 50 wt% white pigment, or 45 wt% white pigment, or 40 wt% white pigment, or 35 wt% white pigment, or 30 wt% white pigment, or 25 wt% white pigment, or 20 wt% white pigment, or 15 wt% white pigment, or 10 wt% white pigment, based on the weight of the composition.
  • a white ink can include 5 wt% to 60 wt%, or 5 wt% to 55 wt%, or 10 wt% to 50 wt%, or 10 wt% to 25 wt%, or 25 wt% to 50 wt%, or 5 wt% to 15 wt%, or 40 wt% to 60 wt% white pigment based on the weight of the composition.
  • a non-white ink can an amount of dye/pigment that is 5%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, 21 wt%, 22 wt%, 23 wt%, 24 wt%, 25%, 26 wt%, 27 wt%, 28 wt%, 29 wt%, 30 wt%, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35%, 36 wt%, 37 wt%, 38 wt%, 39 wt%, 40 wt%, 41 wt%, 42 wt%, 43 wt%, 44 wt%, 45%, 46 wt%
  • the additive or dopant comprises a conductive additive.
  • exemplary conductive additives include, but are not limited to graphite, graphite powder, carbon nanotubes, and metallic particles or nanoparticles, such as gold nanoparticles.
  • the conductive additive is biocompatible and non-toxic.
  • the functionalizing agent is a wound healing agent.
  • a wound healing agent is a compound or composition that actively promotes wound healing process.
  • Exemplary wound healing agents include, but are not limited to dexpanthenol; growth factors; enzymes, hormones; povidon-iodide; fatty acids; anti-inflammatory agents; antibiotics; antimicrobials; antiseptics; cytokines; thrombin; angalgesics; opioids; aminoxyls; furoxans; nitrosothiols; nitrates and anthocyanins; nucleosides, such as adenosine; and nucleotides, such as adenosine diphosphate (ADP) and adenosine triphosphate (ATP); neutotransmitter/neuromodulators, such as acetylcholine and 5-hydroxytryptamine (serotonin/5- HT); histamine and
  • the methods and systems described herein may be deployed in part or in whole through a machine having a computer, computing device, processor, circuit, and/or server that executes computer readable instructions, program codes, instructions, and/or includes hardware configured to functionally execute one or more operations of the methods and systems disclosed herein.
  • the terms computer, computing device, processor, circuit, and/or server, as utilized herein, should be understood broadly.
  • Any one or more of the terms computer, computing device, processor, circuit, and/or server include a computer of any type, capable to access instructions stored in communication thereto such as upon a non-transient computer readable medium, whereupon the computer performs operations of systems or methods described herein upon executing the instructions.
  • such instructions themselves comprise a computer, computing device, processor, circuit, and/or server.
  • a computer, computing device, processor, circuit, and/or server may be a separate hardware device, one or more computing resources distributed across hardware devices, and/or may include such aspects as logical circuits, embedded circuits, sensors, actuators, input and/or output devices, network and/or communication resources, memory resources of any type, processing resources of any type, and/or hardware devices configured to be responsive to determined conditions to functionally execute one or more operations of systems and methods herein.
  • Network and/or communication resources include, without limitation, local area network, wide area network, wireless, internet, or any other known communication resources and protocols.
  • Example and non-limiting hardware, computers, computing devices, processors, circuits, and/or servers include, without limitation, a general purpose computer, a server, an embedded computer, a mobile device, a virtual machine, and/or an emulated version of one or more of these.
  • Example and non-limiting hardware, computers, computing devices, processors, circuits, and/or servers may be physical, logical, or virtual.
  • a computer, computing device, processor, circuit, and/or server may be: a distributed resource included as an aspect of several devices; and/or included as an interoperable set of resources to perform described functions of the computer, computing device, processor, circuit, and/or server, such that the distributed resources function together to perform the operations of the computer, computing device, processor, circuit, and/or server.
  • each computer, computing device, processor, circuit, and/or server may be on separate hardware, and/or one or more hardware devices may include aspects of more than one computer, computing device, processor, circuit, and/or server, for example as separately executable instructions stored on the hardware device, and/or as logically partitioned aspects of a set of executable instructions, with some aspects of the hardware device comprising a part of a first computer, computing device, processor, circuit, and/or server, and some aspects of the hardware device comprising a part of a second computer, computing device, processor, circuit, and/or server.
  • a computer, computing device, processor, circuit, and/or server may be part of a server, client, network infrastructure, mobile computing platform, stationary computing platform, or other computing platform.
  • a processor may be any kind of computational or processing device capable of executing program instructions, codes, binary instructions and the like.
  • the processor may be or include a signal processor, digital processor, embedded processor, microprocessor or any variant such as a co-processor (math co-processor, graphic co-processor, communication co-processor and the like) and the like that may directly or indirectly facilitate execution of program code or program instructions stored thereon.
  • the processor may enable execution of multiple programs, threads, and codes. The threads may be executed simultaneously to enhance the performance of the processor and to facilitate simultaneous operations of the application.
  • methods, program codes, program instructions and the like described herein may be implemented in one or more threads.
  • the thread may spawn other threads that may have assigned priorities associated with them; the processor may execute these threads based on priority or any other order based on instructions provided in the program code.
  • the processor may include memory that stores methods, codes, instructions and programs as described herein and elsewhere.
  • the processor may access a storage medium through an interface that may store methods, codes, and instructions as described herein and elsewhere.
  • the storage medium associated with the processor for storing methods, programs, codes, program instructions or other type of instructions capable of being executed by the computing or processing device may include but may not be limited to one or more of a CD-ROM, DVD, memory, hard disk, flash drive, RAM, ROM, cache and the like.
  • a processor may include one or more cores that may enhance speed and performance of a multiprocessor.
  • the process may be a dual core processor, quad core processors, other chip-level multiprocessor and the like that combine two or more independent cores (called a die).
  • the methods and systems described herein may be deployed in part or in whole through a machine that executes computer readable instructions on a server, client, firewall, gateway, hub, router, or other such computer and/or networking hardware.
  • the computer readable instructions may be associated with a server that may include a file server, print server, domain server, internet server, intranet server and other variants such as secondary server, host server, distributed server and the like.
  • the server may include one or more of memories, processors, computer readable transitory and/or non-transitory media, storage media, ports (physical and virtual), communication devices, and interfaces capable of accessing other servers, clients, machines, and devices through a wired or a wireless medium, and the like.
  • the methods, programs, or codes as described herein and elsewhere may be executed by the server.
  • other devices required for execution of methods as described in this application may be considered as a part of the infrastructure associated with the server.
  • the server may provide an interface to other devices including, without limitation, clients, other servers, printers, database servers, print servers, file servers, communication servers, distributed servers, and the like. Additionally, this coupling and/or connection may facilitate remote execution of instructions across the network. The networking of some or all of these devices may facilitate parallel processing of program code, instructions, and/or programs at one or more locations without deviating from the scope of the disclosure.
  • all the devices attached to the server through an interface may include at least one storage medium capable of storing methods, program code, instructions, and/or programs.
  • a central repository may provide program instructions to be executed on different devices.
  • the remote repository may act as a storage medium for methods, program code, instructions, and/or programs.
  • the methods, program code, instructions, and/or programs may be associated with a client that may include a file client, print client, domain client, internet client, intranet client and other variants such as secondary client, host client, distributed client and the like.
  • the client may include one or more of memories, processors, computer readable transitory and/or non-transitory media, storage media, ports (physical and virtual), communication devices, and interfaces capable of accessing other clients, servers, machines, and devices through a wired or a wireless medium, and the like.
  • the methods, program code, instructions, and/or programs as described herein and elsewhere may be executed by the client.
  • other devices utilized for execution of methods as described in this application may be considered as a part of the infrastructure associated with the client.
  • the client may provide an interface to other devices including, without limitation, servers, other clients, printers, database servers, print servers, file servers, communication servers, distributed servers, and the like. Additionally, this coupling and/or connection may facilitate remote execution of methods, program code, instructions, and/or programs across the network. The networking of some or all of these devices may facilitate parallel processing of methods, program code, instructions, and/or programs at one or more locations without deviating from the scope of the disclosure.
  • all the devices attached to the client through an interface may include at least one storage medium capable of storing methods, program code, instructions, and/or programs.
  • a central repository may provide program instructions to be executed on different devices.
  • the remote repository may act as a storage medium for methods, program code, instructions, and/or programs.
  • the methods and systems described herein may be deployed in part or in whole through network infrastructures.
  • the network infrastructure may include elements such as computing devices, servers, routers, hubs, firewalls, clients, personal computers, communication devices, routing devices and other active and passive devices, modules, and/or components as known in the art.
  • the computing and/or non-computing device(s) associated with the network infrastructure may include, apart from other components, a storage medium such as flash memory, buffer, stack, RAM, ROM and the like.
  • the methods, program code, instructions, and/or programs described herein and elsewhere may be executed by one or more of the network infrastructural elements.
  • the methods, program code, instructions, and/or programs described herein and elsewhere may be implemented on a cellular network having multiple cells.
  • the cellular network may either be frequency division multiple access (FDMA) network or code division multiple access (CDMA) network.
  • FDMA frequency division multiple access
  • CDMA code division multiple access
  • the cellular network may include mobile devices, cell sites, base stations, repeaters, antennas, towers, and the like.
  • the methods, program code, instructions, and/or programs described herein and elsewhere may be implemented on or through mobile devices.
  • the mobile devices may include navigation devices, cell phones, mobile phones, mobile personal digital assistants, laptops, palmtops, netbooks, pagers, electronic books readers, music players, and the like. These mobile devices may include, apart from other components, a storage medium such as a flash memory, buffer, RAM, ROM and one or more computing devices.
  • the computing devices associated with mobile devices may be enabled to execute methods, program code, instructions, and/or programs stored thereon. Alternatively, the mobile devices may be configured to execute instructions in collaboration with other devices.
  • the mobile devices may communicate with base stations interfaced with servers and configured to execute methods, program code, instructions, and/or programs.
  • the mobile devices may communicate on a peer to peer network, mesh network, or other communications network.
  • the methods, program code, instructions, and/or programs may be stored on the storage medium associated with the server and executed by a computing device embedded within the server.
  • the base station may include a computing device and a storage medium.
  • the storage device may store methods, program code, instructions, and/or programs executed by the computing devices associated with the base station.
  • the methods, program code, instructions, and/or programs may be stored and/or accessed on machine readable transitory and/or non-transitory media that may include: computer components, devices, and recording media that retain digital data used for computing for some interval of time; semiconductor storage known as random access memory (RAM); mass storage typically for more permanent storage, such as optical discs, forms of magnetic storage like hard disks, tapes, drums, cards and other types; processor registers, cache memory, volatile memory, non-volatile memory; optical storage such as CD, DVD; removable media such as flash memory (e.g., USB sticks or keys), floppy disks, magnetic tape, paper tape, punch cards, standalone RAM disks, Zip drives, removable mass storage, off-line, and the like; other computer memory such as dynamic memory, static memory, read/write storage, mutable storage, read only, random access, sequential access, location addressable, file addressable, content addressable, network attached storage, storage area network, bar codes, magnetic ink, and the like.
  • RAM random access memory
  • mass storage typically
  • Certain operations described herein include interpreting, receiving, and/or determining one or more values, parameters, inputs, data, or other information.
  • Operations including interpreting, receiving, and/or determining any value parameter, input, data, and/or other information include, without limitation: receiving data via a user input; receiving data over a network of any type; reading a data value from a memory location in communication with the receiving device; utilizing a default value as a received data value; estimating, calculating, or deriving a data value based on other information available to the receiving device; and/or updating any of these in response to a later received data value.
  • a data value may be received by a first operation, and later updated by a second operation, as part of the receiving a data value. For example, when communications are down, intermittent, or interrupted, a first operation to interpret, receive, and/or determine a data value may be performed, and when communications are restored an updated operation to interpret, receive, and/or determine the data value may be performed.
  • the methods and systems described herein may transform physical and/or or intangible items from one state to another.
  • the methods and systems described herein may also transform data representing physical and/or intangible items from one state to another.
  • Example arrangements of programming instructions include at least: monolithic structure of instructions; standalone modules of instructions for elements or portions thereof; and/or as modules of instructions that employ external routines, code, services, and so forth; and/or any combination of these, and all such implementations are contemplated to be within the scope of embodiments of the present disclosure
  • Examples of such machines include, without limitation, personal digital assistants, laptops, personal computers, mobile phones, other handheld computing devices, medical equipment, wired or wireless communication devices, transducers, chips, calculators, satellites, tablet PCs, electronic books, gadgets, electronic devices, devices having artificial intelligence, computing devices, networking equipment, servers, routers and the like.
  • Example hardware includes a dedicated computing device or specific computing device, a particular aspect or component of a specific computing device, and/or an arrangement of hardware components and/or logical circuits to perform one or more of the operations of a method and/or system.
  • the processes may be implemented in one or more microprocessors, microcontrollers, embedded microcontrollers, programmable digital signal processors or other programmable device, along with internal and/or external memory.
  • the processes may also, or instead, be embodied in an application specific integrated circuit, a programmable gate array, programmable array logic, or any other device or combination of devices that may be configured to process electronic signals. It will further be appreciated that one or more of the processes may be realized as a computer executable code capable of being executed on a machine readable medium.
  • the computer executable code may be created using a structured programming language such as C, an object oriented programming language such as C++, or any other high-level or low- level programming language (including assembly languages, hardware description languages, and database programming languages and technologies) that may be stored, compiled or interpreted to run on one of the above devices, as well as heterogeneous combinations of processors, processor architectures, or combinations of different hardware and computer readable instructions, or any other machine capable of executing program instructions.
  • a structured programming language such as C
  • an object oriented programming language such as C++
  • any other high-level or low- level programming language including assembly languages, hardware description languages, and database programming languages and technologies
  • each method described above and combinations thereof may be embodied in computer executable code that, when executing on one or more computing devices, performs the steps thereof.
  • the methods may be embodied in systems that perform the steps thereof, and may be distributed across devices in a number of ways, or all of the functionality may be integrated into a dedicated, standalone device or other hardware.
  • the means for performing the steps associated with the processes described above may include any of the hardware and/or computer readable instructions described above. All such permutations and combinations are contemplated in embodiments of the present disclosure.
  • GO dope in 50% ethanol, 10 mg mL 1 was dry-spun using a custom dry-spinning setup at a rate of 2-5 mm s' 1 using 100 or 200 pm diameter needles. Fiber spools were reduced using a solution of hydriodic acid and ethanol in a 1:4 ratio at 80°C for 4 hours. The spools were rinsed in ethanol three times, dried at 60°C, then stored at room temperature.
  • Hexachloroplatinic acid was added to ethylene glycol to a concentration of lOmM.
  • a 4: 1 molar ratio of sodium hydroxide to platinum salt was formed by adding 1 M NaOH in ethylene glycol to the solution until a concentration of 40mM was reached.
  • a length of rGO fiber was either treated with one pass of a corona wand (Electro-Technic, BD-20AC) or oxygen reactive ion etch (Diener Zepto, 30s, 50W), loosely spooled, then was added to the solution.
  • the corona wand process completed more quickly than the etching and was preferred on the basis of speed.
  • the solution was heated in an oil bath at 110-115 °C for 2 hours, until the solution turns from orange to brown to black.
  • the spool was removed, allowed to cool to room temperature, and then rinsed with ethanol three times. The spool was dried at room temperature overnight.
  • chitosan and silk fibroin solutions were stored at 4°C until used. Lactate oxidase (100 U) was combined with 25 pL silk fibroin (1.0% w/v), 250 pL DI water, 250 pL chitosan (1.0% w/v), and 22 j_iL 1 % glutaraldehyde (0.08M final concentration). The solution was thoroughly mixed and left at room temperature for 20 min. For full-length sensor fibers, 10 cm of PtNP-rGO was cut, then 1.5 cm of the fiber was dip-coated in 1% Nafion (diluted with lx PBS), then dried at 60°C.
  • the fiber was then dip-coated in the enzyme mixture, forming a thick coating (5-10pm) over the Nafion.
  • the layer was dried for 1 hour at room temperature, then the membrane was sprayed with 0.25% PU in THF forming the diffusion limiting layer. The remainder of the sensor was sprayed with 1 % PU in THF.
  • the sensors were stored dry at 4°C until use, up to 2 weeks.
  • CV of PtNP-rGO (scan rate, 50 mV s’ 1 ) was performed in 0.5 M H2SO4 with a salt bridge (100 mM NaSO-i) to prevent chloride contamination from the reference.
  • Hydrogen peroxide sensitivity tests (CV, 50 mV s’ 1 , and CA, 0.5 V vs Ag/AgCl) were performed in 100 mM NaCl and 10 mM phosphate buffer.
  • Electrochemical impedance spectroscopy (EIS) was performed on rGO and PtNP-rGO in 5 mM potassium ferricyanide/ferrocyanide with an ionic background of lOOmM KC1.
  • Lactate sensors were first characterized with CA (0.5V vs. Ag/AgCl) in lOmM phosphate buffer (pH 7.4) with 5 mM additions of lactate. Interference tests were performed in lOmM phosphate buffer with additions of 5mM lactate, 10 mM Na + , 1 mM Mg 2+ , 1 mM Ca 2+ , 1 mM glucose, and 100 pM ascorbic acid. pH sensitivity was measured in 10 mM phosphate-citrate buffer and 10 mM lactate with the pH adjusted between 5.5 and 8 using 1 M HC1 and 1 M NaOH. Temperature sensitivity was measured with the electrochemical cell on a hotplate at 50°C with stirring. The temperature of the solution and current from the sensor in 10 mM lactate and 10 mM phosphate buffer were measured concurrently.
  • the patch was adhered to the lower back, then the readout PCB (in a 3D printed holder) was connected and adhered to the top surface.
  • pH measurements were taken with a skin pH probe (Hanna Instruments, HI99181) to evaluate correlations between the analytical standard (pH probe) and the sensing device here presented.
  • the commercial pH meter was placed near the sensor patch, and readings were taken twice on average during the following: 1) active biking (i.e., “increasing activity” time span): 20 and 25 minutes after the start of the biking session; 2) “cool down” time frame: 3 and 5 minutes after the end of the biking session, which lasted an average of 30 minutes.
  • sweat was collected with filter paper encapsulated by Tegaderm at a location near the sensor patch.
  • the patch was removed, immediately sealed, and replaced twice during biking and once at the end of the “cool down” period. Lactate levels were measured on average during the following time spans: 1) active biking (i.e., “increasing activity” time span): 20 and 25/30 minutes after the start of the exercise session (the latter usually being the end of the session); 2) “cool down” time frame: 5 minutes after the end of the biking session, which lasted an average of 30 minutes.
  • the filter paper was centrifuged at 10,000g, then the collected sweat was diluted down to 8x and 16x and assayed using a colorimetric lactate assay (Sigma MAK329) according to the kit’s SOP.
  • a custom small form factor 4-layer PCB (1.5x2 cm) was developed incorporating a Nordic NRF52832 BLE SOC, a Texas instruments LMP91000 potentiostat, a dual high input impedance operation amplifier (> 1 TQ) and supporting circuitry.
  • the PCB was designed with minimal components, size (limited to the size of the smallest coin cell battery available (CR1216)), and power consumption. During operation it has a low average current draw of 140 pA, allowing for 5 days of continuous measurement with 2 small CR1216 batteries.
  • the output of the LMP91000 (lactate) and voltage buffer (potentiometric) were fed to the analog inputs of the SOC (12-bit SAADC).
  • the sample rate for each channel was 3 Hz with 8x oversampling for noise reduction. Data was postprocessed using a moving average filter with a 15-point window.
  • Acetone, ethanol and hydrochloric acid were of reagent grade or higher quality and obtained from Fisher Scientific. Lactate oxidase was obtained from Toyobo. Conductive silver paint (16062) was purchased from Ted Pella. Deionized water was obtained from a Milli-Q system (18.2 MQ-cm). All chemicals were used without further purification.
  • GO was synthesized from an improved Hummers method using microwave expanded intercalated graphite with methods described previously.
  • the 50% ethanol GO solution was filtered by centrifugation at 1000 g for 10 min, then concentrated to approximately 1.0% by spinning at 10,000 g for 6 hours in a Beckman Coulter Allegra X14R centrifuge. The resulting concentrated GO solution was then stored at 4° C.
  • the solution consisted of single sheets of GO with thicknesses of 0.8-1.2nm and average lateral dimensions of 30 pm, measured via AFM.
  • Spray coating was performed with a gravity feed airbrush. A 0.2mm tip was used for pH and PU membranes, while a 0.5mm tip was used for the reference electrode membrane. Pressure during spraying was approximately 5 psi.
  • Ferrocene functionalized rGO fibers were fabricated as described previously via diazonium coupling of aminoferrocene to GO (See: Napier 2021 ; and Y. Lu, Y. Jiang, H. Wu, W. Chen, Electrochim. Acta 2015, 156, 267). These fibers were found to offer higher capacitance/stability and lower hysteresis as a pH ISE transducer compared to pure rGO fibers (up to 104% increase in capacitance (40 pF for ⁇ 40 pm diameter fiber), measured via polarization ⁇ 1 nA). For the shortterm testing, common to assess wearable configurations for sweat monitoring, unfunctionalized rGO fibers may also perform within the necessary margin of error. Increases in diameter improve capacitance/stability. Additionally, further improvements may include incorporating super hydrophobic conducting polymers with high capacitance, such as PED0T-C14 or POT, as an intermediate transduction layer to reduce the possibility of water layer formation.
  • ISE membrane cocktail was vortexed overnight containing 3.0 mg Hydrogen ionophore II, 100 mg PVC, 197 mg DOS, 2.1 mg KtCPB, and 5 mL of THF. 10 cm lengths of fc-rGO were cut and assembled onto an acrylic frame for spray coating. A 1 cm section of fiber near the bottom of the fiber was spray coated with the cocktail to a thickness of approximately 25 pm. The remainder of the fiber was spray-coated with PU (1% THF) for isolation and stored dry at room temperature until use. Sensors were conditioned overnight in a solution containing 1 mM citric acid, and 1 mM disodium phosphate (pH 3).
  • the reference electrode membrane is a heterogenous polymer membrane acting as a reservoir of solid Ag, AgCl and KC1, silver chloride complexes, and their respective ions. Similar membranes have demonstrated long term stability and insensitivity to ionic background in previous research studies. 10 cm lengths of rGO were cut and assembled onto an acrylic frame. 100 mg of PVC and 15 mg of nitrophenyl octyl ether were dissolved in 5 mL of THF. AgCl and KC1 were mechanically ground in a 1 :5 ratio, then sieved through a 40 pm filter. 300 mg of the powder was added to the solution and vortexed for 5 min.
  • the suspension was sprayed on a 1 cm segment of the rGO fiber near the end of the fiber.
  • the membrane was spray coated with PU (0.25% in THF) forming a thin diffusion-limiting layer (slowing CF diffusion).
  • the remaining fiber was spray coated with PU (1% in THF).
  • the reference electrode was conditioned in 3M KC1 for 5 hours, then stored dry at room temperature. Before assembly into the sensor braid, the reference was conditioned in artificial sweat solution for 30 min to further minimize potential drift.
  • the references showed potentials of -22+5 mV vs. Ag/AgCl (3M).
  • Alternative reference membranes using hydrophobic salts or ionic liquids may offer increased longevity and are being considered for future sensor iterations.
  • rGO fibers were fabricated based on dry-spinning from liquid crystal graphene oxide (GO).
  • GO was produced via an improved Hummers’ method from natural graphite flake (+100 mesh), producing GO sheets with an average lateral size of 30 pm.
  • GO fibers were dry-spun (100-200 pm needles), then reduced in ethanolic hydriodic acid, producing fibers with strong mechanical properties (s 6 GPa, UTS >200 MPa, >9% breaking elongation), and high conductivity (18 kS m 1 ).
  • the multi-sensor “electrochemical yarn” was then developed based on the rGO fibers described above by combining 4-fibers: a three-fiber lactate sensor (i.e. reference, counter, and working electrode fibers) with a pH ion-selective electrode (ISE) fiber, leveraging the same reference fiber used for the lactate sensing unit. These fibers can be braided together to form a yarn that can subsequently be connected to an integrated circuit for signal processing and wireless readout ( Figure 1). The fabrication and operation of the pH and reference electrode fibers were described previously (Napier 2021). The counter electrode and lactate sensor are based on platinum nanoparticle impregnated rGO fibers with the latter also coated with a biopolymer enzymatic layer, both described herein.
  • the rGO fibers Prior to functionalization, the rGO fibers were plasma treated, increasing the surface energy, porosity, and adding surface functional groups - all improving nanoparticle adsorption.
  • grafting functional groups to rGO prior to spinning or platinization may also improve binding and dispersion efficiency as has been demonstrated with rGO and CNTs, albeit with more processing steps.
  • FIG. 13 is a schematic that summarizes the PtNP-rGO fiber preparation.
  • the biosensor detects hydrogen peroxide (H2O2), a byproduct of the enzymatic reaction between lactate oxidase (LOx) in the sensing membrane and lactate in the surrounding solution.
  • LOx converts lactate to pyruvate using oxygen ( Figure 1). Lactate concentrations can be easily correlated to the initial lactate concentration in the biosensor's surroundings, and more importantly, the oxidation of lactate will produce a current that is specific to the target analyte.
  • the sensor shows two linear ranges, the first encompassing three orders of magnitude from 0.2 pM to 400pM of H2O2, the second from 0.5 pM to 5mM of H2O2.
  • the fiber exhibits a high sensitivity of 67.8 ⁇ 0.3 pA mM 1 , or 2.4 mA mM 1 cm' 2 when normalized to the surface area for the first linear range.
  • the high sensitivity can be attributed to the large surface area of dispersed platinum nanoparticles and wrinkled nature of the rGO surface.
  • the PtNP-rGO fiber exhibits high stability with 93.5% of current retained after 14 hours of continuous polarization in 1 mM H2O2 without additional coatings (Figure 6A).
  • the CNT matrix slows the leakage of the soluble, oxidized TTF from the sensor, while allowing for infiltration of the enzymatic coating.
  • Coating rGO fibers with these dispersions may produce sensors with poor sensitivity and stability because of the lack of porosity of conductive supports of this kind (e.g., CNT matrix, carbonaceous fibers). This approach is also difficult to implement in a fiber format, as it may decrease the sensor's flexibility.
  • Solution-based deposition allows for evenly coating large lengths of fibers at a time, enabling rapid sensor fabrication. As compared to electrochemical depositions, this method has increased uniformity along the length of the fiber (gradient deposition due to the resistive fiber) and scalability (spool vs. individual segments).
  • lactate sensors were fabricated on PtNP-rGO fibers using a multi-layer approach (See S. Anastasova, B. Crewther, P. Bembnowicz, V. Curto, H. M. Ip, B. Rosa, G. Z. Yang, Biosens. Bioelectron. 2017, 93, 139) common to platinum-oxidase enzyme systems (Figure 3A).
  • the membranes and coating methods are tailored to increase the linear range of the sensors to cover relevant physiological values, namely using ranges of lactate concentration in sweat between 5 and 30 mM, as the Michaelis-Menten constant (K m ) for LOx (Toboyo) is 1 mM.
  • fibers were coated with a permselective Nafion membrane as an interference rejection layer.
  • the fibers were dip-coated in the active layer containing lactate oxidase (LOx) embedded in a biopolymer matrix consisting of LOx, chitosan, and silk fibroin.
  • LOx lactate oxidase
  • the enzymatic layer needs to be thicker than for other sensing systems, such as glucose.
  • Pre-crosslinking the biopolymer matrix with a low concentration of glutaraldehyde increases the viscosity of the mixture allowing for deposition of a thick layer (-5-10 pm thick) on the fiber in a dip-coating process.
  • the membrane was spray-coated with a layer of polyurethane (PU), acting as a diffusion- limiting layer to further extend the linear range of the sensor into the clinical range for sweat.
  • PU polyurethane
  • Spray-coating was found to improve both speed of fabrication and repeatability of the diffusion layer as multiple dip coatings produced variable thicknesses based on redissolution of the previous layer. It was found that spray coating with 0.25% PU in THF could produce sensors with a linear range up to 30mM, depending on the applied thickness.
  • a yarn consisting of a four- fiber braid is fabricated combining a three-electrode lactate fiber sensor and a pH ion selective (ISE) fiber sensor using a shared reference electrode.
  • the lactate sensor is composed of the PtNP-rGO fiber counter electrode, and an rGO fiber-based reference electrode.
  • the reference electrode membrane is a fiber spray-coated with heterogenous PVC-Ag/AgCl-KCl layer, exhibiting low drift (0.2 mV hr’ 1 ) in artificial sweat with minimal sensitivity to solution composition.
  • the pH sensor is fabricated by spray-coating a polymeric ISE membrane on a ferrocene modified rGO (fc- rGO) fiber for enhanced capacitance/stability (55.8 mV pH’ 1 , Figure 9) (Napier, 2021).
  • the fibers are then interfaced to a socket connector and braided up to the active region (which spans 10 cm in length in each of the fibers) to form the sensor yam.
  • the sensing yarn is subsequently integrated into a wearable patch ( Figure 4A, C) as a demonstrator device.
  • the yam is woven into a rectangle of adsorbent linen fabric and subsequently encapsulating the sensing area between patterned gauze and filter paper, which is then sealed with patterned PET (FLEXcon) with double sided adhesive.
  • FLEXcon patterned PET
  • sweat is channeled through the gauze, then wicked through the filter paper.
  • the wicking of the patch allows for improved temporal measurements in comparison to either direct skin contact or to the use of a single piece of absorbent material (Figure 12).
  • Options are also available for direct integration with microfluidic thread structures, or into more sophisticated microfluidic skin patches.
  • a custom interface to the sensing yarn is built by combining a Bluetooth-based wireless readout circuit (NRF52832 BLE SOC-Nordic Semiconductor - Figure 10) with a commercially available potentiostat chip (LMP91000, Texas Instruments).
  • a custom-written iOS app is used for data collection from the interface.
  • Testing of the sensor patch is performed on two human subjects during exercise on a stationary bike.
  • the output is validated against a commercial pH meter (Hanna) and with chemically assayed sweat samples collected at multiple time points (tested using the Sigma MAK329 for lactate evaluation).
  • the patch is adhered to the lower back of the participant, then the socket interface is connected to the readout integrated front-end. The latter is adhered over the patch during testing as shown in Figure 4A.
  • Data from the sensor is collected during a 30 min cycling workout using the custom iOS application (Figure 4B).
  • a comparison of the data obtained from the yarn with commercial measurements shows high correlation as well as good agreement with expected exercise- induced trends.
  • a calibration correction of -15% accounts for this temperature difference.
  • the lactate data is found to correlate well to the downstream analysis (via commercially available kit) on collected sweat. Data from female participant 2 is shown in Figure 11. Similar high correlation between sensor and probe/assay is found. In this case, the pH values detected are near 7.9 and no corrections were required aside from temperature. Sex, skin-care products, measurement site, and sweat rate are some examples of parameters that affect sweat pH, highlighting the importance of inclusion of calibration elements in the yarn to allow for simultaneous pH sensing and correction of the measured physiological parameters.
  • An electrochemical sensing material comprising: an enzymatic fiber configured to sense a target chemical; at least one of a reference fiber and a counter electrode fiber; wherein the enzymatic fiber and the at least one of the reference fiber and the counter electrode fiber are braided to form a yarn configured for weaving into a patch sensor; and a circuit electrically connected to the yam, wherein the circuit is programmed to record and transmit a sensing signal including one or more physiological parameters sensed by the yarn.
  • enzymatic fiber includes a layer of polyurethane (PU) at least partially coating the biopolymer matrix.
  • PU polyurethane
  • the enzymatic fiber and counter electrode fiber further include platinum nanoparticles (Pt NPs).
  • electrochemical sensing material of any one of the preceding clauses further comprising a processor configured to receive the sensing signal and output one or more physiological parameters sensed by the yam.
  • a method of making an enzymatic fiber comprising: depositing a plurality of metallic nanoparticles onto a conductive fiber, thereby forming a first stage fiber; coating the first stage fiber with a permselective membrane as an interference rejection layer, thereby forming a second stage fiber; coating the second stage fiber with a biopolymer matrix comprising at least one enzyme to form a third stage fiber, wherein the biopolymer matrix is optionally pre-crosslinked prior to coating the second stage fiber; and coating the third stage fiber with a diffusion limiting layer, thereby forming an enzymatic fiber.
  • the second plurality of metallic nanoparticles comprises a metal selected from the group consisting of platinum, silver, gold, palladium, zinc, iron, molybdenum, cobalt, copper, and combinations thereof.
  • a diameter of the electrochemical yarn is between 95 pm and 230 pm, including but not limited to, at least 95 pm, at least 100 pm, at least 120 pm, or at least 150 pm, and at most 230 pm, at most 200 pm, at most 180 pm, or at most 170 pm.
  • a diameter of the first stage fiber is between 20 pm and 50 pm, including but not limited to, at least 20 pm, at least 30 pm, or at least 50 pm, and at most 50 pm, at most 35 pm, or at most 30 pm.
  • a diameter of the second stage fiber is between 25 pm and 60 pm, including but not limited to, at least 25 pm, at least 40 pm, or at least 55 pm, and at most 60 pm, at most 45 pm, or at most 35 pm.
  • a diameter of the third stage fiber is between 30 pm and 70 pm, including but not limited to, at least 30 pm, at least 40 pm, or at least 60 pm, and at most 70 pm, at most 55 pm, or at most 35 pm.
  • a diameter of the enzymatic fiber is between 35 pm and 80 pm, including but not limited to, at least 35 pm, at least 60 pm, or at least 70 pm, and at most 80 pm, at most 65 pm, or at most 45 pm.
  • An enzymatic fiber comprising: a plurality of metallic nanoparticles deposited on a reduced graphene oxide (rGO) fiber; and a biopolymer membrane comprising lactate oxidase disposed about the rGO fiber.
  • rGO reduced graphene oxide
  • a thickness of the biopolymer membrane is between 5 pm and 10 pm, including but not limited to, at least 5 pm, at least 7 pm, or at least 10 pm, and at most 10 pm, at most 7 pm, or at most 5 pm.
  • a diameter of the enzymatic fiber is between 20
  • An electrochemical fiber comprising: a conductive fiber; and a biopolymer membrane coating disposed about the conductive fiber, the biopolymer membrane comprising lactate oxidase, chitosan, and silk fibroin, wherein the biopolymer matrix is optionally pre-crosslinked prior to application to the conductive fiber.
  • a thickness of the biopolymer matrix is between 5 pm and 10 pm, including but not limited to, at least 5 pm, at least 7 pm, or at least 9 pm, and at most 10 pm, at most 7 pm, or at most 6 pm.
  • a diameter of the conductive fiber is between 20 pm and 50 pm, including but not limited to, at least 20 pm, at least 30 pm, or at least 45 pm, and at most 50 pm, at most 35 pm, or at most 30 pm.
  • electrochemical fiber of any one of clauses 47 to 52 further comprising a selectively permeable membrane disposed between the conductive fiber and the biopolymer membrane.
  • a thickness of the selectively permeable membrane is between 5 pm and 10 pm, including but not limited to, at least 5 pm, at least 7 pm, or at least 9 pm, and at most 10 pm, at most 7 pm, or at most 6 pm.
  • a thickness of the diffusion limiting layer is between 5 pm and 10 pm, including but not limited to, at least 5 pm, at least 7 pm, or at least 9 pm, and at most 10 pm, at most 7 pm, or at most 6 pm.
  • the diffusion limiting layer comprises polyurethane.
  • electrochemical fiber of any one of clauses 47 to 57 wherein the electrochemical fiber is configured to be combined with a reference electrode fiber and a counter electrode fiber to form a three-fiber lactate sensor.
  • a diameter of the conductive fiber and the biopolymer membrane is between 25 pm and 60 pm, including but not limited to, at least 25 pm, at least 40 pm, or at least 55 pm, and at most 60 pm, at most 45 pm, or at most 35 pm.
  • a diameter of the conductive fiber, the biopolymer membrane, and the selectively permeable membrane is between 30 pm and 70 pm, including but not limited to, at least 30 pm, at least 40 pm, or at least 60 pm, and at most 70 pm, at most 55 pm, or at most 35 pm.
  • a diameter of the conductive fiber, the biopolymer membrane, the selectively permeable membrane, and the diffusion limiting layer is between 35 pm and 80 pm, including but not limited to, at least 35 pm, at least 60 pm, or at least 70 pm, and at most 80 pm, at most 65 pm, or at most 45 pm.
  • a lactate sensor comprising: an electrochemical yarn, comprising an enzymatic fiber, a reference electrode fiber, and a counter electrode fiber, wherein the enzymatic fiber comprises: a conductive fiber; and a biopolymer membrane comprising lactate oxidase, chitosan, and silk fibroin, wherein the biopolymer matrix is optionally pre-crosslinked prior to application to the conductive fiber.
  • the metallic nanoparticles comprise a metal selected from the group consisting of platinum, silver, gold, palladium, zinc, iron, copper, molybdenum, cobalt, and combinations thereof.
  • a thickness of the diffusion limiting layer or interference rejection layer is between 5 pm and 10 pm, including but not limited to, at least 5 pm, at least 7 pm, or at least 9 pm, and at most 10 pm, at most 7 pm, or at most 6 pm.
  • ISE pH ion-selective electrode
  • a diameter of the pH ISE fiber is between 20 pm and 50 pm, including but not limited to, at least 20 pm, at least 30 pm, or at least 45 pm, and at most 50 pm, at most 35 pm, or at most 30 pm.
  • a thickness of the biopolymer membrane is between 5 pm and 10 pm, including but not limited to, at least 5 pm, at least 7 pm, or at least 9 pm, and at most 10 pm, at most 7 pm, or at most 6 pm.
  • a diameter of at least one of the conductive fiber, reference electrode fiber, or counter electrode fiber is between 20 pm and 50 pm, including but not limited to, at least 20 pm, at least 30 pm, or at least 45 pm, and at most 50 pm, at most 35 pm, or at most 30 pm.
  • a lactate sensor comprising: an electrochemical fiber, comprising: a platinum (Pt) nanoparticle-functionalized (NP-functionalized) reduced graphene oxide (rGO) fiber; an interference rejection layer; a biopolymer membrane comprising lactate oxidase (LOx), chitosan, and silk fibroin, wherein the biopolymer membrane is optionally pre-crosslinked prior to application to the Pt NP-functionalized rGO fiber; and a diffusion limiting layer; and a reference electrode fiber; a counter electrode fiber; and a pH ion-selective electrode (ISE) fiber.
  • Pt platinum
  • NP-functionalized reduced graphene oxide
  • rGO reduced graphene oxide
  • LOx lactate oxidase
  • silk fibroin wherein the biopolymer membrane is optionally pre-crosslinked prior to application to the Pt NP-functionalized rGO fiber
  • LOx lactate oxidase
  • chitosan chitosan
  • a diameter of at least one of the Pt NP-functionalized rGO fiber, reference electrode fiber, or counter electrode fiber is between 20 pm and 50 pm, including but not limited to, at least 20 pm, at least 30 pm, or at least 45 pm, and at most 50 pm, at most 35 pm, or at most 30 pm.
  • a diameter of the lactate sensor is between 95
  • a thickness of at least one of the biopolymer membrane, interference rejection layer, or diffusion limiting layer is between 5 pm and 10 pm, including but not limited to, at least 5 pm, at least 7 pm, or at least 9 pm, and at most 10 pm, at most 7 pm, or at most 6 pm.
  • a diameter of the electrochemical fiber is between 50 pm and 75 pm, including but not limited to, at least 50 pm, at least 60 pm, or at least 70 pm, and at most 75 pm, at most 60 pm, or at most 55 pm.
  • the lactate sensor of any one of clauses 74 to 82 further comprising a circuit interface in electrical communication with the electrochemical yarn, reference electrode fiber, counter electrode fiber, and pH ion-selective electrode (ISE) fiber.
  • ISE pH ion-selective electrode
  • a wearable lactate sensing patch comprising the lactate sensor of any one of clauses 62 to 85.

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  • Yarns And Mechanical Finishing Of Yarns Or Ropes (AREA)

Abstract

L'invention divulgue un matériau de détection électrochimique comprenant une fibre enzymatique configurée pour détecter un produit chimique cible, et une fibre de référence et/ou une fibre de contre-électrode. La fibre enzymatique et la fibre de référence et/ou la fibre de contre-électrode sont tressées pour former un fil configuré pour le tissage dans un capteur de correctif. Le matériau de détection électrochimique comprend également un circuit connecté électriquement au fil, le circuit étant programmé pour enregistrer et transmettre un signal de détection comprenant un ou plusieurs paramètres physiologiques détectés par le fil.
PCT/US2024/014995 2023-02-08 2024-02-08 Fils à détection multiple pouvant être portés pour une surveillance continue sans fil de lactate de sueur Pending WO2024168145A2 (fr)

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US202363483929P 2023-02-08 2023-02-08
US63/483,929 2023-02-08

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CA2999995C (fr) * 2015-09-30 2021-08-24 Siemens Healthcare Diagnostics Inc. Analyseur de fluide pour mesurer des ions de magnesium et procede d'etalonnage de capteur potentiometrique d'ions de magnesium a l'interieur de celui-ci
CN110453260B (zh) * 2019-08-23 2020-06-30 厦门大学 一种用于汗液检测的可穿戴传感器及其制备方法
WO2021232109A1 (fr) * 2020-05-20 2021-11-25 Commonwealth Scientific And Industrial Research Organisation Détection et surveillance de bioanalyte
CN113533474B (zh) * 2021-06-22 2024-04-23 武汉纺织大学 柔性可穿戴电化学生物传感器
CN117858659A (zh) * 2021-07-07 2024-04-09 加利福尼亚大学董事会 可穿戴的非侵入性的微针传感器
WO2023247956A1 (fr) * 2022-06-21 2023-12-28 Sm24 Ltd Capteurs biométriques

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