CN115558600A - A mechanical loading device for 3D artificial blood vessel culture - Google Patents

Abstract

The invention discloses a mechanical loading device for 3D artificial blood vessel culture, belonging to the field of microbiology devices; the inner end of the hollow shaft of the first rotary slip ring and slip ring is connected with a first hollow connecting piece through a hollow shaft of a stepping motor of a hollow shaft stepping motor, and the first hollow connecting piece penetrates through the culture cavity and is sealed and fixed with one end of the artificial blood vessel in the culture cavity; the inner end of the hollow shaft of the second rotary slip ring and the other end of the artificial blood vessel in the culture cavity are sealed and fixed through a second hollow connecting piece; the peristaltic pump, the flask, the second rotating slip ring, the second hollow connecting piece, the artificial blood vessel, the hollow shaft stepping motor, the first hollow connecting piece and the first rotating slip ring form a sealed loop together; a peripheral resistance generating device is arranged on a hose line between the flask and the second rotary slip ring; the invention can realize the control of the flow of the pipeline and the pressure difference in the pipeline by adjusting the rotating speed of the stepping motor, and simulate the pulsating flow of the human body.

Description

A mechanical loading device for 3D artificial blood vessel culture

technical field

The invention belongs to the technical field of microbiological devices, in particular to a mechanical loading device for 3D artificial blood vessel culture.

Background technique

The invention relates to a bioreactor mechanical loading device in the field of biological 3D printing. Vascular transplantation, as the best treatment for major cardiovascular diseases, has received extensive attention at home and abroad. Affected by factors such as the number of donations and the low success rate of matching due to immune rejection, the shortage of donors required for vascular transplantation has always been very large, and it is difficult to meet the clinical needs. As a new technology closely connected with tissue engineering and regenerative medicine, 3D bioprinting has developed rapidly in the past decade, bringing new solutions to the problem of implantable organ donor gap.

3D printing technology has a wide range of applications in tissue engineering and regenerative medicine due to its personalized manufacturing characteristics. Blood vessels are relatively simple tissues and organs in the human body. There have been many literature reports at home and abroad on the application of bio-3D printing technology to artificial angioplasty, involving extrusion 3D printing, inkjet 3D printing and UV-assisted 3D printing. Extrusion bio-3D printing is currently the most widely used bio-3D printing technology in the field of artificial blood vessels, but its printing resolution is relatively low. Inkjet 3D printing has a higher printing resolution, which solves the problem of low resolution of extrusion bio-3D printing, but due to the small diameter of the nozzle, it cannot well complete the printing task of higher-viscosity bio-ink. UV-assisted 3D printing combines the advantages of the former two, which can print high-precision three-dimensional structures, but ultraviolet rays can damage cells. Artificial blood vessel printing technologies based on extrusion bio-3D printing technology include: concentric circle overlay printing method, embedded angioplasty method, suspension angioplasty method, coaxial printing blood vessel formation method, rotating rod-assisted forming method, etc.

As the main device for the subsequent functional induction of the printed structure of 3D printed artificial blood vessels, the bioreactor provides a stable and similar culture environment for in vitro tissues/organs. In the process of artificial blood vessel preparation, in vitro culture and maturation of printed artificial blood vessels is an extremely important link, and tissue engineering bioreactors are usually used to provide them with the required culture environment. At present, the function of bioreactors in the construction of tissue/organ mechanical environment in vitro is relatively single, mainly to provide sterile environment, carbon dioxide concentration, temperature regulation and nutrient solution for tissue and organ culture, but cannot fully simulate the tissue/organ in vivo Mechanics environment.

Therefore, there is an urgent need for a 3D printing platform suitable for biological blood vessels, which is used for the study of functional induction of artificial blood vessels during in vitro culture.

Contents of the invention

Aiming at the problems existing in the background technology, the present invention provides a mechanical loading device for 3D artificial blood vessel culture, which is characterized in that it includes: a hose line, a first rotating slip ring, a hollow shaft stepping motor, a first hollow Connecting piece, culture chamber, sealing cover, second hollow connecting piece, second rotating slip ring, electric displacement platform, peripheral resistance generating device, flask, peristaltic pump and artificial blood vessel, wherein the first rotating slip ring and hollow shaft stepping motor The bracket is installed on the main platform, and the inner end of the hollow shaft of the first rotating slip ring is connected with the first hollow connecting piece through the hollow shaft of the stepping motor of the hollow shaft stepping motor, and the first hollow connecting piece passes through the culture chamber and its One end of the internal artificial blood vessel is sealed and fixed; the inner end of the second rotating slip ring slip ring hollow shaft is sealed and fixed with the other end of the artificial blood vessel inside the culture cavity through the second hollow connector; the outer end of the first rotating slip ring slip ring hollow shaft The end is connected to the outer end of the peristaltic pump, the flask, the second rotary slip ring slip ring hollow shaft in turn through the hose line, the peristaltic pump, the flask, the second rotary slip ring, the second hollow connecting piece, the artificial blood vessel, the hollow shaft stepper The motor, the first hollow connector, and the first rotating slip ring together form a sealed circuit; a peripheral resistance generating device is installed on the hose path between the flask and the second rotating slip ring;

An electric displacement platform is arranged on the side of the hollow shaft stepping motor object of the main platform, and the second rotating slip ring is installed on the electric displacement platform through a bracket. The reciprocating movement direction of the electric displacement platform is parallel to the axial direction of the artificial blood vessel inside the culture chamber , the moving displacement platform and the main platform are fixed on the loading platform on the test bench; the culture chamber is filled with culture fluid, and the liquid level of the culture fluid just reaches the lower edge of the artificial blood vessel.

The electric displacement platform is connected with the tension and pressure loading module in the control system, the hollow shaft stepping motor is connected with the torsional force loading module in the control system, the peripheral resistance generating device, the hollow shaft stepping motor and the peristaltic pump are connected with the control system The pulse force loading module is connected, and the first rotating slip ring, the hollow shaft stepping motor and the second rotating slip ring are connected with the rotation angle maintaining module in the control system.

A rotation angle sensor is provided at the second hollow connector, and the rotation angle sensor is connected to the rotation angle maintenance module; whenever a feedback signal from the hollow shaft stepping motor is received, the rotation angle maintenance module sends a reverse signal of the same rotation angle to the first rotary slip ring At the same time, whenever the rotation angle sensor collects a rotation angle signal that satisfies one unit, the rotation angle maintenance module sends it to the second rotating slip ring for reverse rotation.

The rotation angle maintaining module sends a signal to the second rotating slip ring to rotate in the same direction when the hollow shaft stepper motor is started and braked.

The first hollow connecting part and the second hollow connecting part are kept sealed with the culture cavity.

The connection form of the artificial blood vessel between the second hollow connecting piece and the first hollow connecting piece is: the two ends of the artificial blood vessel are respectively sleeved outside the second hollow connecting piece and the first hollow connecting piece and fixed by a parafilm.

The inner diameters of the slip ring hollow shaft of the second rotary slip ring, the slip ring hollow shaft of the first rotary slip ring, the hollow shaft of the stepping motor, the first hollow connector and the second hollow connector are equal to the inner diameter of the artificial blood vessel.

The beneficial effects of the present invention are:

1. Through three modules, fully simulate various problems that may be encountered in artificial blood vessels (also called hydrogel tubes) or cytoskeleton, forming a complete data collection chain.

2. The peristaltic pump is used to provide pulsating pressure for the blood vessels. After the motor driver receives the control command, the stepping motor starts to drive the pump head to rotate. Every time the stepping motor rotates once, several round shafts on the pump head squeeze the soft pipe in turn, and the pump out the liquid. Therefore, adjusting the speed of the stepping motor can realize the control of the pipeline flow and the control of the pressure difference in the pipeline, simulating the pulsating flow of the human body.

3. Both the stepping motor shaft and the rotating slip ring shaft are designed to be hollow and have the same aperture, so that the best simulated artificial blood vessel can always obtain sufficient oxygen and nutrients, and discharge metabolic waste in time in the actual working state.

Description of drawings

Fig. 1 is a schematic flowchart of an embodiment of a mechanical loading device for 3D artificial blood vessel culture in the present invention.

Fig. 2 is a schematic oblique view of the vicinity of the culture chamber in the embodiment of the present invention.

Among them, 1-hose line, 2-pneumatic joint, 3-first rotary slip ring, 4-hollow shaft stepper motor, 5-first hollow connector, 6-culture chamber, 7-sealing cover, 8-the first Two hollow connectors, 9-second rotating slip ring, 10-electric displacement platform, 11-peripheral resistance generating device, 12-flask, 13-peristaltic pump, 14-artificial blood vessel, 15-hollow shaft of stepper motor, 16- Parafilm.

detailed description

The present invention will be further described in detail below in conjunction with the accompanying drawings.

The embodiment of the present invention shown in Fig. 1 and Fig. 2 includes: a hose line 1, a pneumatic joint 2, a first rotary slip ring 3, a hollow shaft stepping motor 4, a first hollow connecting piece 5, a culture chamber 6, Sealing cover 7, second hollow connector 8, second rotating slip ring 9, electric displacement platform 10, peripheral resistance generating device 11, flask 12, peristaltic pump 13 and artificial blood vessel 14, wherein first rotating slip ring 3 and hollow shaft The stepping motor 4 is supported on the main platform (not marked in the figure), and the inner end of the first rotating slip ring 3 slip ring hollow shaft passes through the hollow shaft of the stepping motor 4 and the first hollow connector 5 connected, the first hollow connector 5 passes through the culture chamber 6 and one end of the artificial blood vessel inside is sealed and fixed; the inner end of the second rotating slip ring 9 slip ring hollow shaft passes through the second hollow connector 8 and the artificial The other end of the blood vessel 14 is sealed and fixed; the top of the culture chamber 6 is equipped with a sealing cover 7; the outer end of the first rotating slip ring 3 and the hollow shaft of the slip ring connect with the peristaltic pump 13, the flask 12, and the second rotating slip ring through the hose line 1 in turn. The outer end of the ring 9 slip ring hollow shaft is connected, the peristaltic pump 13, the flask 12, the second rotary slip ring 9, the second hollow connector 8, the artificial blood vessel 14, the hollow shaft stepper motor 4, the first hollow connector 5, The first rotary slip ring 3 together forms a sealed circuit; the hose line 1 between the flask 12 and the second rotary slip ring 9 is equipped with a peripheral resistance generating device 11; DMEM culture solution is added to the culture chamber so that its liquid level just reaches The lower edge of the artificial blood vessel 14 eliminates the influence of gravity on the blood vessel and avoids the leakage problem caused by the excessively high liquid level;

The hollow shaft stepper motor 4 object side of the main platform is provided with an electric displacement platform 10, and the second rotating slip ring 9 is installed on the electric displacement platform 10 by a support, and the reciprocating movement direction of the electric displacement platform 10 is the same as that of the cultivation cavity 6 inside. The axial directions of the artificial blood vessels are parallel, and the moving displacement platform 10 and the main platform are fixed on the loading platform on the test bench.

The control system includes: tension and pressure loading module, torsional force loading module, pulse force loading module and rotation angle maintenance module;

The electric displacement platform 10 is connected with the tension and pressure loading module; by controlling the axial movement of the electric displacement platform 10 along the artificial blood vessel, the artificial blood vessel is reciprocally stretched and compressed to apply tension and pressure to it, and the second rotating slider installed on the electric displacement platform 10 The ring 9 is connected to the second hollow connector 8 through threaded connection (the specific position is G1/8 pipe thread), and one end of the artificial blood vessel is fixed on the second hollow connector 8 by a parafilm.

The hollow shaft stepper motor 4 is connected to the torsional force loading module, and one end of the artificial blood vessel is clamped and fixed on the first hollow connector 5 by the sealing film. The stepping motor hollow shaft of the stepping motor drives the first hollow connecting piece 5 to rotate, and applies torque to the artificial blood vessel.

The peripheral resistance generating device 11, the hollow shaft stepping motor 4 and the peristaltic pump 13 are connected to the pulsating force loading module that sends the pulsating flow, and the tension and pressure loading module and the torque module are used as part of the pulsating flow hose line. When pumping the pulsating force, the peristaltic The pump core rotates with the stepping motor shaft; then the radius of the hose line 1 is changed by using the peripheral resistance generating device 11 to generate different peripheral resistances. The left port of the peristaltic pump is connected with the first rotary slip ring through the hose line 1 through the pneumatic plug, the right port of the peristaltic pump is connected with the left port of the two flat-bottomed flasks through the hose line 1, and another hose line is drawn from the right port of the flask 1. Passing through the outer peripheral resistance generating device 11, the hose line 1 is connected with the hollow shaft of the slip ring on the second rotating slip ring through a pneumatic joint (thread at one end, bayonet at the other end).

The first rotary slip ring 3, the hollow shaft stepping motor 4 and the second rotary slip ring 9 are connected to the rotation angle maintenance module, and a rotation angle sensor is provided at the second hollow connector 8, and the rotation angle sensor is connected to the rotation angle maintenance module; When the rotation signal of the hollow shaft stepping motor 4 is received, the rotation angle maintaining module sends a reverse signal of the same rotation angle to the first rotating slip ring 3;

When the hollow shaft stepping motor 4 is started and braked, the rotation angle maintenance module sends a signal to the second rotating slip ring 9 to perform a slight rotation in the same direction as for the artificial blood vessel (usually no more than two step angle multiples), In this way, the stepper motor of the left torsion force module can play a negative feedback role in the unstable stage of experiment startup and braking, so that the external pipeline and printing blood vessels are affected by the fluctuation of the system to a minimum;

The rotation angle maintenance module is also equipped with a monitoring function. Whenever the rotation angle sensor collects a rotation angle signal that meets a unit (the unit here is usually the step angle multiple of the used rotary slip ring and hollow shaft stepper motor), the rotation angle The maintenance module is sent to the two rotating slip rings 9 for reverse rotation, thereby ensuring the integrity of soft components such as the hose line 1 and the artificial blood vessel 14 .

In this embodiment, the first rotary slip ring 3 and the second rotary slip ring 9 are hydraulic components, and it is necessary to consider that the maximum pressure threshold must conform to the pulsating pressure range of 10.7-16.0kPa in the human body, so slip rings are used; and Both choose the single-channel rotary slip ring of the MEPH100-02 (L0305) model of Mofulon Technology Co., Ltd. In order to make the connection between the rotary slip ring and the hollow shaft stepping motor and the pneumatic joint (threads at both ends) have good sealing performance, R1/8 is selected as the threaded connection in combination with the size of the shaft diameter; at the same time, in order to reduce the weight, the main components are made of stainless steel and aluminum alloy Used in combination, the biocompatibility of the rotary slip ring is guaranteed.

In this embodiment, in order to make the nutrient solution pass through the rotating slip ring and avoid the turbulence problem caused by the change of pipe diameter, a through hole with a diameter of 5mm is opened inside the slip ring hollow shaft of the rotating slip ring and the artificial blood vessel (elastic ) single channel with the same inner diameter, and the left and right sides use the same specification of rotating slip rings, so that the entire flow channel maintains a consistent inner diameter; similarly, the inner diameter of the hollow shaft of the stepping motor is also 5mm, and the hollow shaft of the two slip rings and the hollow shaft of the stepping motor , The inner diameters of the first hollow connecting part 5 and the second hollow connecting part 8 are consistent with the inner diameter of the artificial blood vessel.

In this embodiment, both the first hollow connecting piece 5 and the second hollow connecting piece 8 are kept sealed with the culture cavity 6, and the sealing form is connected with the pneumatic joint through a pipe thread, so that the culture cavity is in a relatively independent state, so as to be connected The inlet hose line 1 will not be entangled and interfered when the modules work together to ensure that the caliber of the silicone tube will not change due to twisting or stretching, thereby maintaining the stability of the overall system;

In this embodiment, the hose line 1 is a BPT hose line;

In this embodiment, the material of the main platform is an acrylic plate;

In this embodiment, the hollow shaft of the hollow shaft stepping motor 4 is provided with a sleeve outside the hollow shaft of the stepping motor;

In this embodiment, the peripheral resistance generating device 11 is a three-jaw chuck.

When working, the tensile and pressure loading modules, torsional loading modules and pulse loading modules in the system respectively drive their respective components,

The tension and pressure loading module connected to the electric displacement platform 10 performs tension simulation through the movement of the second rotating slip ring 9 along the axial direction of the artificial blood vessel 14;

The torsional force loading module connected to the hollow shaft stepping motor 4 performs torsion loading. The hollow shaft stepping motor 4 rotates according to the demand, and at the same time, the first hollow connecting part 5 rotates in the opposite direction at an equal rotation angle. During this process, all the hoses Neither the road 1 nor the second hollow connector 8 rotates;

The pulse force loading module connected with the peripheral resistance generating device 11, the hollow shaft stepping motor 4 and the peristaltic pump 13, the stepping motor is pulsed by the drive board, rotates at a specified angle, and the pump core of the peristaltic pump rotates with the stepping motor shaft, Pumping pulsating force; then the pulsating flow is simulated by the periodic operation of the peristaltic pump 13 and the peripheral resistance generating device 11;

When the three modules are in operation, the rotation angle maintenance module monitors to ensure that the hose line 1 and artificial blood 14 are intact during long-term automatic work, thereby ensuring effective data collection.

Claims (7)

1. A mechanical loading device for 3D artificial blood vessel culture, which is characterized by comprising: the device comprises a soft pipeline (1), a first rotary sliding ring (3), a hollow shaft stepping motor (4), a first hollow connecting piece (5), a culture cavity (6), a sealing cover (7), a second hollow connecting piece (8), a second rotary sliding ring (9), an electric displacement platform (10), a peripheral resistance generating device (11), a flask (12), a peristaltic pump (13) and an artificial blood vessel (14), wherein the first rotary sliding ring (3) and the hollow shaft stepping motor (4) are both arranged on a main platform in a supporting mode, the inner end of a sliding ring hollow shaft of the first rotary sliding ring (3) is connected with the first hollow connecting piece (5) through the hollow shaft of the stepping motor of the hollow shaft stepping motor (4), and the first hollow connecting piece (5) penetrates through the culture cavity (6) and is sealed and fixed with one end of the artificial blood vessel inside the first rotary sliding ring (3); the inner end of the slip ring hollow shaft of the second rotary slip ring (9) is hermetically fixed with the other end of the artificial blood vessel (14) in the culture cavity (6) through a second hollow connecting piece (8); the outer end of a slip ring hollow shaft of the first rotary slip ring (3) is sequentially connected with the outer ends of a peristaltic pump (13), a flask (12) and a slip ring hollow shaft of the second rotary slip ring (9) through a soft pipeline (1), and the peristaltic pump (13), the flask (12), the second rotary slip ring (9), a second hollow connecting piece (8), an artificial blood vessel (14), a hollow shaft stepping motor (4), a first hollow connecting piece (5) and the first rotary slip ring (3) form a sealed loop together; a peripheral resistance generating device (11) is arranged on the hose line (1) between the flask (12) and the second rotary slip ring (9);

an electric displacement platform (10) is arranged on one side of a hollow shaft stepping motor (4) object of the main platform, a second rotary slip ring (9) is installed on the electric displacement platform (10) through a bracket, the reciprocating direction of the electric displacement platform (10) is parallel to the axial direction of the artificial blood vessel in the culture cavity (6), and the motion displacement platform (10) and the main platform are both fixed on a loading platform of the experiment table; the culture cavity (6) is filled with culture solution, and the liquid level of the culture solution is flush with the lower edge of the artificial blood vessel (14).

2. The mechanical loading device for 3D artificial blood vessel culture according to claim 1, wherein the electric displacement platform (10) is connected to a tension and pressure loading module in a control system, the hollow shaft stepping motor (4) is connected to a torsion loading module in the control system, the peripheral resistance generating device (11), the hollow shaft stepping motor (4) and the peristaltic pump (13) are connected to a pulsating force loading module in the control system, and the first rotating slip ring (3), the hollow shaft stepping motor (4) and the second rotating slip ring (9) are connected to a rotation angle maintaining module in the control system.

3. The mechanical loading device for 3D artificial blood vessel culture according to claim 2, characterized in that a rotation angle sensor is arranged at the second hollow connecting piece (8), and the rotation angle sensor is connected with a rotation angle maintaining module; when receiving a feedback signal of the hollow shaft stepping motor (4), the rotation angle maintaining module sends a reverse signal with the same rotation angle to the first rotating slip ring (3), and when collecting a rotation angle signal meeting one unit, the rotation angle maintaining module sends the rotation angle signal to the second rotating slip ring (9) to rotate reversely.

4. Mechanical loading device for 3D artificial blood vessel culture according to claim 2 or 3, characterized in that the rotation angle maintaining module sends signals to the second rotating slip ring (9) to rotate in the same direction when the hollow shaft stepping motor (4) is started and stopped.

5. Mechanical loading device for 3D artificial blood vessel culture according to claim 1, characterized in that the first hollow connector (5) and the second hollow connector (8) are kept sealed with the culture chamber (6).

6. The mechanical loading device for 3D artificial blood vessel culture according to the claim 1, characterized in that the connection form of the second hollow connecting piece (8) and the artificial blood vessel (14) of the first hollow connecting piece (5) is: two ends of the artificial blood vessel are respectively sleeved outside the second hollow connecting piece (8) and the first hollow connecting piece (5) and are fixed by a sealing film.

7. The mechanical loading device for 3D artificial blood vessel culture according to claim 1, characterized in that the inner diameters of the slip ring hollow shaft of the second rotary slip ring (9), the slip ring hollow shaft of the first rotary slip ring (3), the stepping motor hollow shaft, the first hollow connector (5) and the second hollow connector (8) are equal to the inner diameter of the artificial blood vessel.

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