WO2015161539A1

WO2015161539A1 - Medical biological tissue structure, preparation method therefor and special apparatus thereof

Info

Publication number: WO2015161539A1
Application number: PCT/CN2014/078238
Authority: WO
Inventors: 王小红; 刘利彪
Original assignee: 清华大学
Priority date: 2014-04-21
Filing date: 2014-05-23
Publication date: 2015-10-29
Also published as: CN103892937A; CN103892937B

Abstract

A medical biological tissue structure, a preparation method therefor and a special apparatus thereof. The medical biological tissue structure includes a hollow tube (801), a functional layer (802) attached to the hollow tube and a synthetic macromolecular protective film (803), wherein the hollow tube (801) is made of biological materials containing cells or without cells, and the functional layer (802) contains or does not contain cells. On the basis of the attachment cross-linking and solidity principle, the hollow tube (801) is firstly extruded out through a special spraying nozzle under the control of a computer, then the biological materials are sprayed and attached to the hollow tube (801) to form the functional layer (802), and then are accumulated layer by layer to form a composite molding body, and finally a synthetic macromolecular solution is sprayed to the outer surface of the composite molding body to form the protective film (803). According to the set molding steps, a three-dimensional structure body containing synthetic macromolecular materials, cells and natural biological materials and having a complicated space shape and a porous structure is finally manufactured. The medical biological tissue structure can be molded at normal temperature, the process is simple, the cell survival rate is high, the cell distribution is even and controllable, and the medical biological tissue structure has a good mechanical property and a good biological property.

Description

Medical biological tissue structure, preparation method thereof and special equipment

The invention belongs to the technical field of tissue and organ manufacturing, and particularly relates to a medical biological tissue structure, a preparation method thereof and a special equipment.

Background technique

Bulk tissue (such as bone, cartilage, breast, muscle, skin) and internal organs (such as heart, liver, spleen, lung, kidney, etc.) are important components of the human body, related damage caused by disease, trauma and aging. Dysfunction, a serious hazard to human health and quality of life. Traditional clinical methods still have a number of limitations, especially those that do not allow for the controlled distribution of cells in vitro. The cross-integration of manufacturing science and life sciences has led to the development of cell-controlled assembly technology, a new way to directly construct organ replacements with new generations of new functions in vitro.

The Low Temperature Deposition Manufacturing Process (LOV) is a new process developed by the Institute of Materials Processing Technology of Tsinghua University for the special requirements of biomaterial forming. The low-temperature deposition manufacturing means that the scaffold material is made into a liquid state, and the solution is extruded in a filament form through a head, and is deposited in a low-temperature forming chamber.

The specific process of low temperature deposition manufacturing generally adopts the following process:

1) Create a 3D model using 3D modeling software, layer the model with layered processing software, and obtain coordinate codes for forming.

2) Select the materials of the experiment, prepare the solution according to the appropriate ratio, and make it for use.

3) The material is added to the injector of each nozzle of the forming apparatus, and the control software in the computer controls the scanning motion and the extrusion and ejection movement of each nozzle according to the input layer file and the set processing parameters. In the low temperature forming chamber, the material emerging from the nozzle is rapidly solidified and bonded to each other to form a frozen stent.

4) The frozen stent is placed in a freeze dryer, subjected to freeze-drying treatment, and the solvent is removed to obtain a stent which is solid at normal temperature. During this process, sublimation of the solvent produces a microporous structure within the frozen scaffold.

At present, the Department of Mechanical Engineering of Tsinghua University has a corresponding three-dimensional stent controlled molding device with single nozzle and double nozzle, and has made research on the design and forming performance of the nozzle, and designed and fabricated the piston extrusion nozzle. For example, CLRF-2000-II biomaterial rapid prototyping machine independently developed by Tsinghua University Advanced Manufacturing Rapid Prototyping Laboratory.

However, complex tissues or organs in the human body are generally composite structures composed of two or more different cells and extracellular matrix materials, and the structures are interconnected. With the deepening of research, the requirements for the formation of three-dimensional structures of heterogeneous materials are proposed. The original single nozzle and double nozzle cannot meet the requirements for rapid manufacturing of complex tissues and organs; Chinese Patent Document (Application No. 201 1 1 0205970. 1) relates to a fixed multi-head three-dimensional controlled forming system for complex organ precursors, and a spraying device For two fixed motor booster nozzles, the forming table is arranged on the three-dimensional motion device, different nozzle components are equipped with different forming materials; all the nozzles are in the same plane, and the three-dimensional motion device makes the working nozzle and the forming table when switching the nozzle Positive. The stepping motor is fixed on the Z-direction motion device bracket, and the screw can exert a certain pressure on the forming material in the nozzle under the driving of the linear stepping motor, and the forming material is then sprayed from the nozzle, and the heating rod and the heat insulating jacket are mounted on the nozzle. The lower section maintains the forming material in the spray head at a set temperature.

This design is simple and reliable. However, the fixed multi-nozzle forming system has the following shortcomings: 1) Low temperature deposition requires a low temperature environment of minus ₄₀ degrees. Low temperature will adversely affect the cell survival rate, and it is necessary to add a cryopreservation agent. The process is more complicated. 2) The original system has only two nozzles, that is, two materials can be used, but the complex tissues and organs contain a variety of materials and cells, and the existing equipment cannot satisfy the manufacture of complex tissue organ precursors. 3) The concentration of the cells inside the formed body is not uniform, and the concentration of the cells is relatively high, and the individual cells cannot be accurately located and encoded. 4) The side surface of the molded body cannot be sprayed. The motor-assisted rapid prototyping head needs to be installed vertically. If the horizontal installation is performed, the slurry will drip from the nozzle and will fall under the action of its own gravity. The side surfaces are attached.

Summary of the invention

The invention aims at the deficiencies of the prior art, and provides a medical biological tissue structure, a preparation method thereof and a special device. The invention is based on the adhesion cross-linking curing technology, so as to realize the accuracy of various materials and cells under normal temperature environment. Forming and accurate positioning of individual cells further enhance the flexibility of shaping and the accuracy of cell distribution.

The technical solution of the present invention is as follows:

A medical biological tissue structure, characterized in that: the medical biological tissue structure comprises a layer-by-layer stacked hollow tube with or without cells, a functional layer with or without cells adhered to the hollow tube, and a synthetic The polymer protective film; the hollow tube is distributed inside the protective film, and the two ports of the hollow tube protrude out of the protective film; the functional layer is a natural polymer aqueous solution or hydrogel with or without cells And forming a composite formed body with the hollow tube; the protective film is located outside the composite formed body, and is a synthetic polymer material; the hollow tube is formed by mixing and curing the natural polymer solution and the crosslinking agent, wherein the natural polymer The solution contains or does not contain cells.

In the above technical solution, each of the hollow tubes in the layer-by-layer stacked hollow tubes is in a quadrangular, serpentine, wavy or circular shape arranged in parallel, or a personalized structure is designed according to clinical needs; 01〜0. 1 The protective film is a porous structure, and the film thickness is preferably 0. 01〜0. 1 The protective film is a porous structure, and the film thickness is preferably 0. 01〜0. 01~20nm

The invention also provides a special device for preparing the medical biological tissue structure, the device comprising a bottom plate, a Y-direction moving device, an X-direction moving device, a Z-direction moving device, a rotating forming station, a high-pressure gas source, a control unit, and a plurality of nozzles; the Y-direction moving device is mounted on the bottom plate, the X-direction moving device is mounted on the Y-direction moving device, and the rotating forming table is mounted on the top of the X-direction moving device, wherein: the device further comprises an arc with meshing teeth The plurality of spray heads include a protective film spray head, a first functional layer spray head, a second functional layer spray head, and a hollow tube forming spray head mounted on the Z-direction moving device; the curved rail is mounted on the bottom plate, the curved shape The central axis of the track coincides with the central axis of the Y-direction moving device; three sliders with meshing teeth driven by the stepping motor are mounted on the curved track, the protective film nozzle, the first functional layer nozzle and the second function The layer nozzles are mounted on the three sliders by hangers. A special device for preparing a medical biological tissue structure, characterized in that: the hollow tube forming nozzle comprises an A component syringe, a B component syringe and a nozzle; a crosslinking chamber and a central shaft are arranged in the nozzle, A The component syringe and the B component syringe are in communication with the crosslinking chamber.

The utility model relates to a special device for preparing a medical biological tissue structure, characterized in that: the second functional layer nozzle and the protective film nozzle are pneumatic spray valves, and the structure comprises an inner pipe and an outer sleeve, and the pneumatic spray valve passes through the gas pipeline and The high pressure gas source is connected.

A special device for preparing a medical biological tissue structure, characterized in that: the first functional layer nozzle comprises a rack, a solution storage box, a screening device and a piezoelectric spray valve; a solution storage box, a screening device and a piezoelectric spray valve The screening device is installed on the pylon; the screening device is installed under the solution storage box and connected through the pipeline; the screening device comprises a screening plate and a substrate (502), and a screening groove is drawn on one side of the substrate, on both sides of the front side of the screening groove A screening board is installed.

The invention provides a method for preparing the medical biological tissue structure, characterized in that the method comprises the following steps: 1) under the control of the control unit, the X-direction moving device and the Y-direction moving device drive the rotating forming table to move to the middle The set position below the empty tube forming nozzle starts the hollow tube forming nozzle, and the natural polymer solution and the crosslinking agent in the hollow tube forming nozzle are mixed and extruded to form one or a set of hollow tube structures arranged in parallel;

2) Rotating the forming table is driven by the X-direction moving device and the Y-direction moving device, moves to the set position below the curved track, activates the first functional layer nozzle, and sprays the biological material with or without cells to the hollow tube. Surface; then initiating a second functional layer nozzle, spraying a second or no cell-containing biological material onto the surface of the hollow tube, forming a layer containing the plurality of biological materials and cells on the surface of the hollow tube structure formed in step 1) Functional layer

3) The Z-direction moving device drives the hollow tube forming nozzle to move, and the parallel hollow tubes formed in the step 1) are vertically pressed to press another set of parallel-arranged hollow tubes, and the steps 2) are repeated to form a composite formed body;

4) repeating the above steps 1), 2), 3) to form a composite formed body of a multilayer structure;

5) The protective film nozzle is activated, and the synthetic polymer solution is sprayed in a mist form to form a protective film on the surface of the composite molded body to produce a tissue organ precursor having a spatially complex shape and a composite material.

In the method of the present invention, the natural polymer solution and the crosslinking agent in the step 1) are respectively a sodium alginate solution and an aqueous calcium chloride solution, or a fibrinogen and a thrombin solution, respectively, or They are respectively a collagen solution and a cell culture medium solution, or a polylactic acid glycolic acid solution and water, respectively, or a polyurethane solution and water, respectively, or a calcium hydrogen phosphate dihydrate and calcium hydroxide solution, respectively, or a monohydrated phosphoric acid. a solution of calcium hydride and calcium oxide, or a solution of tetracalcium phosphate and calcium hydrogen phosphate, respectively, or a solution of calcium phosphate solution and polylactic acid glycolic acid, or a solution of hydroxyapatite water and P polylactic acid glycolic acid, respectively; The concentration of the sodium alginate solution and the calcium chloride solution is: the sodium alginate is dissolved in the cell culture medium, the mass concentration is 0.1 - 5% (w / v), the calcium chloride aqueous solution is in deionized water, The mass concentration is 1 - 10% (w / v); the concentration of fibrinogen and thrombin solution is: the fibrinogen powder is dissolved in the cell culture medium solution, the mass concentration is 0.01-5% (w/v), the thrombin is dissolved in deionized water at a concentration of 50-200 Unt; the concentration of the collagen solution is: The collagen is dissolved in the cell culture medium at a concentration of 0.01-5. /. (w/v) _{; the} concentrations of calcium hydrogen phosphate dihydrate and calcium hydroxide solution are respectively: mixing calcium hydrogen phosphate and calcium hydroxide in a ratio of 1-20% (w/v) by mass with 1M disodium hydrogen phosphate solution; The concentrations of the monohydrate calcium dihydrogen phosphate and the calcium oxide solution are respectively: The monohydrate calcium dihydrogen phosphate and the calcium oxide particles are divided into mass concentrations of 1-20. /. (w/v) The ratio is mixed with 1M phosphate buffer; the mass concentration of tetracalcium phosphate and calcium hydrogen phosphate solution are: 1-20% (w/v) of phosphate tetracalcium phosphate and calcium phosphate phosphate respectively. Mixing; polylactic acid glycolic acid solution and polyurethane solution concentration are: polylactic acid glycolic acid, polyurethane dissolved in tetraethylene glycol, mass concentration is 0.1-15<1⁄2: w/v) _; the organism described in step 2) Materials include cells, growth factors, cell cryopreservation, anticoagulant materials, drugs, gelatin, hyaluronic acid, collagen, silk fibroin, laminin, elastin, monosaccharide, disaccharide, dextrose, viscous One or several combinations of polysaccharide, heparin, chitosan, phosphorylated chitosan, sulfated chitosan and matrigel, wherein the cell density is 1 X10 ² /ΓΤ1_-1 Χ10 ⁷ /rrl_, The cells are adult cells, such as one or a combination of osteoblasts, hepatocytes, cardiomyocytes, stellate cells, fibroblasts, embryonic stem cells, induced pluripotent stem cells, and adipose stem cells, and the growth factor concentration is 10- 50ng/rrL, cell cryopreservation body Aqueous solution of dimethyl sulfoxide, glycerol or a solution, or dextrose solution, concentrations are 1-20. 3⁄4^, anticoagulant material, drug, gelatin, hyaluronic acid, collagen, silk fibroin, laminin, elastin, monosaccharide, disaccharide, dextrose, mucopolysaccharide, heparin, chitosan, phosphorylation The mass concentration of chitosan, sulfated chitosan and matrigel is 0.1-20. 3⁄4w/v) _; The synthetic polymer solution described in the step 3) dissolves the synthetic polymer in an organic solvent, and the synthetic polymer is polyurethane, or polylactic acid glycolic acid, or polyethylene, or polypropylene, or poly One or a combination of caprolactone, or polycarbonate, or polyethylene glycol, or polyhydroxy acid ester, the organic solvent is synthesized using dimethyl sulfoxide, tetraethylene glycol or 1.4 dioxane The mass concentration of the polymer solution is 1-30% (w/v).

The invention has the following advantages and outstanding effects: 1) The invention adopts an adhesive cross-linking curing forming method, can realize various materials at room temperature forming, can greatly improve the survival rate of cells, and make a plurality of cells fixedly distributed in different reservations. The position, relative to low-temperature deposition manufacturing, avoids complicated and expensive refrigeration equipment, eliminates the use of cryopreservation reagents such as cryopreservation agents, and greatly simplifies the forming process. 2) The invention uses multiple sets of spraying devices to work together, can spray a variety of materials and cells, and multiple sets of spraying devices can move independently with each other, eliminating mutual interference between multiple sets of spraying devices, and greatly reducing equipment. volume of. 3) The present invention employs a combination of specific pneumatic extrusion, spray coating and piezoelectric spraying, wherein the pneumatic spraying has high precision and fast response speed. And the spray valve sprays the spray liquid after atomization, and the contact area between the liquid and the air is increased, the solvent can be quickly volatilized to improve the forming efficiency, and the sprayed cells can be reliably combined with the existing surface, and the single layer of cells can be sprayed. The spray width is large and the spraying efficiency is high. Piezoelectric spraying can also be sprayed in a spot for precise spraying to achieve precise positioning and encoding of cells. The combination of the two is that the distribution of cells in the structure is more uniform, and the cell spraying technique is quantified and accurate, and the randomness is reduced. 4) The invention has multi-degree of freedom motion, can accurately process the circle and the ring cross section, and the relative angle between the central axis of the nozzle and the surface of the rotary forming table can also be changed, and the side surface of the forming body can be sprayed, which is convenient and complicated. The manufacture of curved surfaces.

DRAWINGS

1 is a schematic view of a medical biological tissue structure provided by the present invention.

Fig. 2 is a schematic diagram showing the three-dimensional structure of a special device for preparing a medical biological tissue structure. Fig. 3 is a schematic view showing the appearance of a hollow tube forming nozzle.

Fig. 4 is a schematic view showing the internal structure of a hollow tube forming nozzle.

Figure 5 is a schematic view showing the structure of the first functional layer nozzle.

Figure 6 is a schematic view showing the structure of a screening device.

Figure 7 is a schematic view of the structure of the spray valve.

Fig. 8 is a schematic view showing the electrical control of a medical biological tissue structure-dedicated device of the present invention.

Figure 9 is a process flow diagram of the process of the present invention.

In the figure: 101-protective film nozzle; 102-second function layer nozzle; 103-first functional layer nozzle; 104-curved track; 105-X-direction moving device; 106-rotating forming table; 107-Y-direction moving device 108- bottom plate; 109-Z direction moving device; 110- hollow tube forming nozzle; 111-high pressure gas source; 112-electric cabinet; 113-control unit; 301-A component syringe; 302-B component syringe; 303-crosslinking chamber; 304-central shaft; 303-nozzle; 401-hanger; 402-solution storage box; 403-screening device; 404-piezoelectric spray valve; 501-screening plate; 502-substrate; Groove; 601-inlet; 602-internal tubing; 603-inlet; 604-outer sleeve; 801-hollow tube; 802-functional layer; 803-protective film.

Specific actual 51^

In order to further understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.

1 is a schematic view of a medical biological tissue structure provided by the present invention, the medical biological tissue structure comprising a layer-by-layer stacked hollow tube 801 containing or not containing cells, adhered to or contained on the hollow tube The functional layer 802 of the cell and the synthetic polymer protective film 803; the hollow tube 801 is distributed inside the protective film 803, and the two ports of the hollow tube 801 extend out of the protective film; the functional layer is included or not a cell-containing natural polymer aqueous solution or hydrogel, and a composite formed body with the hollow tube 801; the protective film 803 is located outside the composite molded body, and is a synthetic polymer material; the hollow tube 801 is composed of a natural polymer solution and The crosslinking agent is formed by mixing and crosslinking, wherein the natural polymer solution may or may not contain cells. Each of the hollow tubes 801 of the layer-by-layer stacked hollow tubes 801 is arranged in a quadrangular shape, a serpentine shape, a wave shape or a circular shape, or a personalized structure according to clinical needs; adjacent two layers of hollow tubes are arranged in a cross arrangement The inner diameter of the hollow tube is generally 0.01 5, and the thickness of the functional layer 802 with or without cells is 0.01 0.1. The protective film 803 has a porous structure and the film thickness is 0.01 20 .

2 is a schematic diagram of a three-dimensional structure of a special apparatus for preparing a medical biological tissue structure, which comprises a protective film nozzle 101, a second functional layer nozzle 102, a first functional layer nozzle 103, and an arc-shaped track 104 X-direction movement. The apparatus 105 rotates the forming table 106, the Y-moving device 107, the bottom plate 108, the Z-direction moving device 109, the hollow tube forming nozzle 110, the high-pressure gas source 111, the electrical cabinet 112, and the control unit 113. The curved track 104Y to the moving device 107 Z to the moving device 109, the high pressure gas source 111 and the electrical cabinet 112 are all mounted on the bottom plate 108. Wherein, the curved track 104 is mounted at the front end of the bottom plate 108, the Y-direction moving device 107 is mounted at the central axis position of the bottom plate 108, and the Z-direction moving device 109 is mounted at the rear end of the bottom plate 108 opposite to the Y-direction moving device 107, and Y To the center line and curved track of the moving device 107 104. The center line of the Z-direction moving device 109 is in the same plane; the high-pressure gas source 1 1 1 and the electrical cabinet 1 12 are installed side by side at the rear end position of the bottom plate 108.

The present invention enables multi-degree of freedom motion, including three linear motions in the X-direction, Y-direction, and Z-direction, as well as rotational motion about the Z-direction rotation. The X-direction moving device 105 is fixedly mounted on the top of the Y-direction moving device 107, and the rotary forming table 106 is mounted on the top of the X-direction moving device 105.

The invention has a total of four spraying devices installed, and the four spraying devices are respectively a hollow tube forming nozzle 110, a first functional layer nozzle 103, a second functional layer nozzle 102 and a protective film nozzle 101, between the four spraying devices. Exercise independently of each other. The hollow tube forming nozzle 1 10 is mounted on the slider of the Z-direction moving device 109, and the first functional layer head 103, the second functional layer head 102, and the protective film head 101 are mounted side by side on the curved track 104.

The appearance of the hollow tube forming nozzle 1 10 is as shown in Fig. 3. The hollow tube forming nozzle 110 is mounted on the slider of the Z-direction moving device 109, which is combined with the X-direction moving device 105, the Y-direction moving device 107, and the rotary forming. The coordinated movement of the table 106 can form a complex three-dimensional structure.

Hollow tube forming nozzle 1 10 The internal structure is shown in Fig. 4. The structure of the hollow tube forming nozzle 1 10 includes an A-component syringe 301, a B-component syringe 302, a crosslinking chamber 303, a central shaft 304, and a nozzle 305.

The A-component syringe 301 and the B-component syringe 302 are respectively provided with different pre-crosslinking solutions, and the two solutions are mixed and crosslinked in the crosslinking chamber 303 to form a gel state of a certain mechanical property, and then the gelled substance formed by crosslinking is formed. Extrusion by the nozzle 305 under the push of air pressure, because the central position of the nozzle is mounted with the central shaft 304, at which point the extrudate forms a hollow tubular structure.

The structure of the first functional layer showerhead 103 is as shown in Fig. 5. The structure of the first functional layer showerhead 103 includes a pylon 401, a solution storage cartridge 402, a screening device 403, and a piezoelectric injection valve 404.

The solution storage case 402, the screening device 403, and the piezoelectric injection valve 404 are both fixed to the slider of the curved track 104 by being mounted on the pylon 401, thereby achieving the purpose of moving on the curved track.

The aqueous solution containing the cells is contained in the solution storage case 402, and the solution is screened as it flows through the screening device 403, and the cells containing the cells are sprayed through the piezoelectric injection valve 404. The screening device 403 is mounted below the solution storage box 402, and the piezoelectric injection valve 404 is mounted below the screening device 403, and the three are connected by a pipe.

The screening device 403 increases the content of cells in the spray droplets and reduces the probability of spraying without cell droplets. The structure is shown in Fig. 6, and includes a screening plate 501, a substrate 502, and a screening tank 503. The material of the substrate 502 is plastic or glass, and the groove is formed on the surface thereof to form a liquid-passing screening groove 503; the screening plate 501 is made of a copper plate, and two copper plates are a group, and the screening plate 501 is installed at the beginning of the screening groove 503. Because the droplets containing cells have a certain charge polarity, the alternating current of the positive voltage on the two copper plates will form an electric field of a certain frequency, so that the liquid containing the cells will be deflected by the electromagnetic force, and the liquid containing no cells. The droplets flow freely without being affected by the electromagnetic force.

The piezo spray valve ₄₀₄ can spray a small amount of droplets to achieve precise spraying of individual cells.

The second functional layer nozzle 102 and the protective film nozzle 101 are both pneumatic spray valves, and the structure thereof is as shown in FIG. 7 , and includes an inner pipe 602 and an outer sleeve 604 . The difference between the two is that the liquid to be sprayed is different, and the second functional layer The sprayed solution of the spray head is a cell-containing solution to form a cell layer 802, and the solution sprayed by the protective film spray head 101 is a cell-free polymer solution to form a protective film 803 on the outer surface of the structure. The top of the inner pipe 602 is a liquid inlet 601, and the solution to be sprayed enters the nozzle through the liquid inlet 601; the side wall of the outer sleeve 604 is opened with an air inlet 603, and the filtered high-pressure gas enters the nozzle through the air inlet 603. The liquid is broken up at the spray of the spray head to form a mist.

The high-pressure gas used in the present invention is generated by a high-pressure gas source 11 1 , and the high-pressure gas source 11 1 and the shower head are connected by a gas path, and the gas path structure includes an air compressor, a pressure gauge, a gas storage tank, a cooler, The filter and the nozzle controller, wherein the air compressor, the pressure gauge, the gas storage tank, the cooler, and the filter are all installed inside the casing of the high pressure gas source 1 1 1 . The air compressor generates high-pressure air, and then the high-pressure air is sent to the storage tank for storage. The gas storage tank not only has the function of storing compressed gas, but also reduces the fluctuation of the pressure of the compressed gas. The pressure gauge shows the gas pressure value in the gas tank. The high-pressure air sent from the gas storage tank has a very high temperature, and needs to be cooled by the cooler to a temperature value that meets the working requirements, and then a filter is used to thoroughly filter the water, oil and other impurity particles in the high-pressure air. . The high pressure gas that meets the requirements after filtration through the filter is divided into two paths. One of the channels is connected to the spray solution syringe, and the high pressure gas provides pressure to the liquid in the spray solution syringe. The other high-pressure gas is connected to the nozzle controller installed inside the electrical cabinet 12, and the high-pressure gas from the nozzle controller is connected to the air inlet of the nozzle to control the start and stop of the nozzle.

The X-direction motion device 1 05, the Y-direction motion device 1 07 and the Z-direction motion device 1 08 are composed of a ball screw pair, a linear guide, a slider, a coupling, and a stepping motor. Wherein, the two ends of the ball screw and the linear guide are respectively fixed on the sliders on the left and right sides of the Y-direction moving device 1 08, and the stepping motor is coupled through the coupling and the ball screw.

The three nozzles mounted on the curved track can be moved to different positions and cooperate with the rotary forming table 105. The relative angle between the central axis of the nozzle and the surface of the rotary forming table 105 can be changed. The surface is sprayed to facilitate the manufacture of complex surfaces.

The electrical cabinet 1 12 is equipped with a control driver for each motion device and a nozzle controller for the pneumatic nozzle. They are connected to the control unit 1 13 via electrical lines.

The control unit 1 13 provides a user-friendly user interface, analyzes and processes the 3D file to be formed, outputs correct motion and spray start and stop commands, compensates for mechanical deviations, calibrates the working state of the nozzle and test equipment. The spraying device set under the control of the control unit 1 13 starts to accurately position the material to be sprayed, so that various materials, including high viscosity gels, slurries, solutions, and low viscosity cell-containing solutions, The polymer solution is sprayed at a set spatial position. Three-dimensional controlled assembly of two or more cell and scaffold materials is achieved.

The control circuit of the special equipment used for the medical biological tissue structure of the present invention is as shown in FIG. 8. Using the CAN bus control mode, the seven stepping motors used in the present invention are connected to the bus through the control driver to receive the control unit 1 13 . The control signal, such as the stepping motor of the X-direction device 105, is connected to the bus through the X-direction control driver; the nozzle controller is connected to the bus through the I/O controller, receives the signal of the control unit 1 13, and then passes through the gas path. On and off to control the start and stop of the nozzle.

The product flow processed by the present invention is as shown in FIG. 9. The hollow tube 801 is extruded from the hollow tube forming nozzle 110, and the first functional layer nozzle 103 and the second functional layer nozzle are equipped with different cell solutions. 1 02 ejects the corresponding cell solution to form a functional layer 802 on the surface of the hollow tube 801, and then the protective film nozzle 101 sprays the polyurethane or PLG liquid to the periphery of the formed hepatocyte layer to form the outermost protective film 803 . The working process of the present invention will be described below with reference to FIG. 9 and the following embodiments:

The materials of the experiment were selected and arranged in a suitable ratio to form a forming material for use.

The natural polymer solution and the crosslinking agent are respectively a fibrinogen and a thrombin solution. The fibrinogen solution is prepared by dissolving fibrinogen powder in an OVEM solution at a mass concentration of 0.1. /. (w/v), Thrombin was dissolved in deionized water at a concentration of 100 U/rrL. The adipose stem cells are mixed into the fibrinogen solution at a density of 1×10 ⁶ /rrl_. The endothelial cell growth factor (50 ng/rrL) was added thereto, and the fibrinogen solution containing cell and endothelial cell growth factor and the thrombin solution were separately placed in two different syringes of the hollow tube forming nozzle 110 for use.

Functional cells such as hepatocytes and stellate cells are extracted from the patient. The above cells are mixed with a natural polymer fibrinogen solution. The density of hepatocytes is 1 X 10 ⁷ / rrL, and the density of stellate cells is 1 X 10 ³ / rrl_. The cell-containing polymer solution is separately charged into the corresponding syringe for use.

The polyurethane was dissolved in tetraethylene glycol to a solution having a concentration of 5% (w/v), and was placed in a spray solution syringe of the protective film nozzle 101 for use.

The rotary forming table 106 is moved to the set position below the hollow tube forming nozzle under the driving of the X-direction moving device 105 and the Y-direction moving device 107, and the hollow tube forming nozzle 110 extrudes the hollow tube 801 containing cells and growth factors. Then, the X-moving device 105 and the Y-moving device 107 are driven to rotate the forming table 106 to a position set below the curved track 104, and the first functional layer head 103 and the second functional layer head 102 are provided with different cell solutions. The corresponding cell solution is sprayed on the surface of the hollow tube to form a functional layer 802, and the polyurethane solution is sprayed onto the periphery of the formed hepatocytes and the star-shaped functional layer 802 by a protective film nozzle 101 equipped with a spray valve to form an outermost layer. The protective film 803 of the layer achieves mechanical properties that match the mechanical properties of the hepatic arteries and venous vessels.

The 3D model of the liver blade was established by using 3D modeling software, and the model was layered by layered processing software to obtain the NC code for forming, and the layer file and processing parameters were input into the computer control software.

The initial coordinates of the X-direction moving device 105, the Y-direction moving device 107, the Z-direction moving device 109, and the rotary forming table 106 are set.

The movement parameter of the movement mechanism is controlled by the control unit 113 according to the input machining parameters, and the rotary forming table 106 is moved by the X-direction movement device 105 and the Y-direction movement device 107 to the set position below the hollow tube forming nozzle 110, and then The hollow tube forming nozzle 110 starts to work, and the hollow tube 801 extruded from the head is solidified and formed in a normal temperature environment, and layers are stacked. As the accumulation of each layer is completed, the hollow tube forming nozzle 110 is raised by the Z-direction moving device 109.

If it is necessary to process the ring-like structure, under the control of the control unit 113, the moving mechanism drives the optional forming table 106 to the designated position, starts the rotary forming table 106, and rotates the forming table 106 around its central axis, and so on. After one rotation of the table 105, the Z-moving device 107 drives the hollow tube forming nozzle 110 to raise the set height to form a continuous shape.

According to the setting procedure, after the intermediate tubular structure is formed, the rotary forming table 106 is moved by the Y-direction moving device 107 to a set position below the curved track 104, and the first functional layer head 103 is activated on the curved track 104. Moving to the set position, the cell-containing solution is sprayed onto the surface of the tubular structure 801; then, the second functional layer showerhead 102 is activated, spraying another type of cell solution at a specific location of the tubular structure, and continuing controlled forming. Finally, the rotary forming table 106 is kept rotated, and the polyurethane or PL solution is sprayed onto the periphery of the formed functional layer 802 by the protective film nozzle 101 equipped with the spray valve to form the outermost protective film 803.

After the different materials are formed on the upper surface of the rotary forming table 106, the rotary forming table 106 is moved out of the forming processing area in accordance with the procedure, the forming process is finished, and then the formed structure can be taken out.

Example 2: The natural polymer solution and the crosslinking agent were a sodium alginate solution and a calcium chloride solution. The sodium alginate solution has a concentration of 5% (w/v) and the calcium chloride solution has a concentration of 1% (w/v). The endothelial cells were mixed into a sodium alginate solution at a density of 1 X 10 ⁵ / rrl_. The sodium alginate solution containing the endothelial cells and the calcium chloride solution were separately placed in two different component syringes of the hollow tube forming nozzle 1 10 for use.

Cardiomyocytes and Schwann cells are extracted from the patient. The above cells were mixed with a sodium alginate solution. The myocardial cell density is 1 X 10 ⁶ / rrl_, and the Schwann cell density is 1 X 10 ⁴ / rrl_. The cell-containing polymer solution is separately charged into the corresponding syringe for use.

The PL was dissolved in tetraethylene glycol solution to a concentration of 20. /. The solution of (w/ v) is placed in the spray solution syringe of the protective film nozzle 101 for use.

The rotary forming table 106 is moved by the X-direction moving device 105 and the Y-direction moving device 107 to a set position below the hollow tube forming nozzle, and the hollow tube forming nozzle 110 extrudes a hollow tube containing cells and growth factors. Structure 801; Then, the X-moving forming device 106 and the Y-moving device 107 are driven to rotate the forming table 106 to a position set below the curved track 104, and the first functional layer head 103 and the second functional layer are provided with different cell solutions. The nozzle 102 is sprayed with a corresponding cell solution adhered to the surface of the hollow tube 801 to form a functional layer 802. The polyurethane solution is sprayed onto the periphery of the formed functional layer 802 by a protective film nozzle 101 equipped with a spray valve to form an outermost layer. The protective film 803 achieves mechanical properties that match the mechanical properties of the heart arteries and venous vessels.

The control unit 1 13 controls the motion parameters of the motion mechanism according to the input machining parameters, and the rotary forming table 106 is moved by the X-direction motion device 105 and the Y-direction motion device 107 to the set position below the hollow tube forming nozzle 1 10 . Then, the hollow tube forming nozzle 1 10 starts to work, and the hollow tube 801 extruded from the nozzle is solidified and formed in a normal temperature environment, and layers are stacked. As the accumulation of each layer is completed, the hollow tube forming nozzle 1 10 is raised by a Z-direction moving device 109 by a set height.

If it is necessary to process the ring-like structure, under the control of the control unit 136, the moving mechanism drives the optional forming table 106 to the designated position, starts the rotary forming table 106, and rotates the forming table 106 around its central axis, and so on. After one rotation of the forming table 105, the Z-moving device 107 drives the hollow tube forming nozzle 1 10 to raise the set height to form a continuous shape.

According to the setting procedure, after the middle hollow tube 801 is formed, the rotary forming table 106 is moved by the Y-direction moving device 107 to a set position below the curved track 104, and the first functional layer head 103 is activated in the curved track. The upper portion 104 is moved to the set position, and the cell-containing solution is sprayed onto the surface of the tubular structure 801; then, the second functional layer head 102 is activated, and another type of cell solution is sprayed at a specific position of the hollow tube 801 to continue controlled forming. Finally, the rotary forming table 106 is kept rotating, and the protective film nozzle 101 is activated to spray the PLG liquid to the periphery of the formed cell-containing functional layer 802 to form the outermost protective film 803.

The number and type of nozzles mounted on the curved track 104 can be interchanged, including pneumatic nozzles, motor boosters or piezoelectric nozzles, all of which can squeeze or eject the solution in a filament shape. In addition, a plurality of sets of spraying devices can be integrated and fixedly mounted on the Z-direction moving structure 109, and simultaneously moved under the control of the control unit 1 13 , but only one set of spraying devices squeezes and ejects materials at a time.

Example 3: The natural polymer solution and the crosslinking agent were a solution of calcium hydrogen phosphate dihydrate (DCPD) and calcium hydroxide (1 M disodium hydrogen phosphate). The weight percentages of DCPD and calcium hydroxide in 1 M disodium hydrogen phosphate solution were 20% and 10% (w/v), respectively. The DCPD and calcium hydroxide solutions were separately placed in two different component syringes of the hollow tube forming nozzle 1 10 for use.

Osteoblasts and adipose stem cells are extracted from the patient. The above cells were mixed with a 5% (w/v) gelatin (PBS) solution. The density of osteoblasts was 1 10 ⁵ / and the density of adipose stem cells was 1 10 ² /. Adding endothelial cell growth factor d Ong/ rrL) The cell-containing polymer solution was separately filled into the corresponding syringe for use.

The PU was dissolved in a tetraethylene glycol solution to prepare a solution having a concentration of 10% (w/v), which was placed in a spray solution syringe of the protective film nozzle 101 for use.

The rotary forming table 106 is moved by the X-direction moving device 105 and the Y-direction moving device 107 to a set position below the hollow tube forming nozzle 110, and the hollow tube forming nozzle 110 is extruded into cells and growth factors. The empty tube 801; then, the X-moving device 105 and the Y-moving device 107 are driven to rotate the forming table 106 to a position set below the curved track 104, the first functional layer nozzle 103 and the second function with different cell solutions. The layer nozzle 102 sprays a corresponding cell solution to adhere to the surface of the hollow tube to form a functional layer 802 containing a cell layer, and sprays a polyurethane solution onto the periphery of the formed functional layer by a protective film nozzle 101 to form an outermost protective film. 803 can be connected to an artery or a venous blood vessel.

The control unit 1 13 controls the motion parameters of the motion mechanism according to the input machining parameters, and the rotary forming table 106 is moved by the X-direction motion device 105 and the Y-direction motion device 107 to the set position below the hollow tube forming nozzle 1 10 . Then, the hollow tube forming nozzle 1 10 starts to work, and the hollow tube 801 extruded from the nozzle is solidified and formed in a normal temperature environment, and layers are stacked. As the accumulation of each layer is completed, the hollow tube forming nozzle 1 10 is raised by a specific height by the Z-direction moving device 109.

When processing the ring-like structure, under the control of the control unit 136, the moving mechanism drives the optional forming table 106 to the designated position, starts the rotary forming table 106, and rotates the forming table 106 about its central axis, and so on. After one rotation of the stage 105, the Z-moving device 107 drives the hollow tube forming nozzle 1 10 to raise the set height to form a continuous shape.

According to the setting procedure, after the middle hollow tube 801 is formed, the rotary forming table 106 is moved by the Y-direction moving device 107 to a set position below the curved track 104, and the first functional layer head 103 is activated in the curved track. 104 moves to the set position, spraying the cell-containing solution onto the surface of the hollow tube 801; then, the second functional layer nozzle 102 is activated, in the hollow tube Another type of cell solution was sprayed at 801 specific locations and continued controlled formation.

Finally, the rotary forming table 106 is kept rotating, and the PU solution is sprayed onto the periphery of the formed functional layer 802 by the protective film nozzle 101 equipped with the spray valve to form the outermost protective film 803.

Example 4: The natural polymer solution and the crosslinking agent were a polylactic acid glycolic acid (PL) solution and water, respectively. The weight percentage of PL in 1.4 dioxane is 20% (w/v), respectively. The PLG liquid and water are separately charged into the two different component syringes of the hollow tube forming nozzle 1 10 for use.

5% (w / v) gelatin (PBS), 1% (w / v) fibrinogen solution was mixed, respectively, added to 0.01. /. (w/ v) Heparin and osteoblast growth factor were placed in the corresponding syringes for later use.

The PU was dissolved in a tetraethylene glycol solution to prepare a solution having a concentration of 20% (w/v), which was placed in a spray solution syringe of the protective film nozzle 101 for use.

The rotary forming table 106 is moved by the X-direction moving device 105 and the Y-direction moving device 107 to a set position below the hollow tube forming nozzle, and the hollow tube forming nozzle 110 extrudes the hollow tubular structure 801; then, X The rotating forming table 106 is moved to the set position below the curved track 104 to the moving device 105 and the Y-moving device 107, and the first functional layer head 103 and the second functional layer head 102 are filled with different polymer solutions, and are ejected. The corresponding polymer solution adheres to the surface of the hollow tube 801 to form a functional layer 802 of a multi-layered biomaterial structure, and the polyurethane solution is sprayed onto the periphery of the formed functional layer 802 by a protective film nozzle 101 equipped with a spray valve to form the outermost layer. A protective film 803 of the layer.

According to the setting procedure, after the middle hollow tube 801 is formed, the rotary forming table 106 is moved by the Y-direction moving device 107 to a set position below the curved track 104, and the first functional layer head 103 is activated in the curved track. The upper portion 104 is moved to the set position, and the cell-containing solution is sprayed onto the surface of the hollow tube 801; then, the second functional layer head 102 is activated, and another type of cell solution is sprayed at a specific position of the hollow tube 801 to form a functional layer 802.

Finally, the rotary forming table 106 is kept rotating, and the PU solution is sprayed onto the periphery of the formed cell-containing functional layer 802 by a protective film nozzle 101 equipped with a spray valve to form an outermost protective film 803.

After the different materials are formed on the upper surface of the rotary forming table 106, the rotary forming table 106 is moved out of the forming processing area in accordance with the procedure, the forming process is completed, and then the formed structure can be taken out. In practical applications, the number and type of nozzles mounted on the curved track 104 can be exchanged, including pneumatic nozzles, motor boosters or piezoelectric nozzles, all of which can squeeze or eject the solution in a filament shape. In addition, multiple sets of spraying devices can also be integrated and fixedly mounted on the Z-direction moving structure 109, and simultaneously moved under the control of the control unit 1 13, but only one set of spraying devices squeezes and ejects materials at a time.

The invention relates to a medical biomaterial assembly device and method based on adhesion cross-linking curing, which has the advantages that: a plurality of materials and a plurality of cells (including single cells) can be spatially positioned in a normal temperature environment. Accurate positioning, the realization of complex curved surface manufacturing and spraying, greatly improve the cell survival rate and simplify the forming process.

The present invention has been described in detail with reference to the preferred embodiments of the present invention. All modifications, equivalents, improvements, etc., within the spirit and scope of the present invention are intended to be included within the scope of the present invention. , and not limited to the above embodiments.

Claims

What is claimed is: 1. A medical biological tissue structure, characterized in that: the medical biological tissue structure comprises a layer-by-layer stacked hollow tube (801) containing or not containing cells, and a cell containing or not adhering to the hollow tube The functional layer (802) and the synthetic polymer protective film (803); the hollow tube (801) is distributed inside the protective film (803), and the two ports of the hollow tube (801) protrude out of the protective film; The functional layer is a natural polymer aqueous solution or hydrogel with or without cells, and forms a composite formed body with the hollow tube (801); the protective film (803) is located outside the composite formed body, and is highly synthetic. The molecular material; the hollow tube (801) is formed by mixing and curing the natural polymer solution and the crosslinking agent, wherein the natural polymer solution contains or does not contain cells.

2. A medical biological tissue structure according to claim 1, wherein each of said hollow tubes (801) stacked one by one is in parallel, quadrangular, serpentine, wavy. Or round, or according to clinical needs to design a personalized structure; adjacent two layers of hollow tubes are arranged in a cross, hollow tube inner diameter is 0.01~5

3. A medical biological tissue structure according to claim 1 or 2, wherein: the thickness of the functional layer (802) with or without cells is 0.01 to 0. Imrt

4. The medical biological tissue structure according to claim 1 or 2, wherein: the protective film (803) has a porous structure, and the film thickness is 0.01 to 20 nm.

5. A special device for preparing the medical biological tissue structure according to claim 1, the device comprising a bottom plate (108), a Y-direction moving device (107), an X-direction moving device (105), and a Z-direction moving device (109) a rotary forming station (106), a high pressure gas source (111), a control unit (113), and a plurality of nozzles; the Y-direction moving device (107) is mounted on the bottom plate (108), and the X-direction moving device (105) is mounted on On the Y-direction moving device (107), the rotary forming table (106) is mounted on the top of the X-direction moving device (105), characterized in that: the device further comprises an arc-shaped track (104) with meshing teeth, the plurality of The nozzle includes a protective film nozzle (101), a first functional layer nozzle (103), a second functional layer nozzle (102), and a hollow tube forming nozzle (110) mounted on the Z-direction moving device (109); (104) mounted on the base plate (108), the central axis of the curved track (104) coincides with the central axis of the Y-direction moving device (107); the curved track (104) is mounted with three stepper motors Slider with meshing teeth, said protective film spray The head (101), the first functional layer showerhead (103) and the second functional layer showerhead (102) are respectively mounted on the three sliders by a pylon.

6. A special device for preparing a medical biological tissue structure according to claim 5, wherein: said hollow tube forming nozzle (110) comprises an A component syringe (301) and a B component syringe (302). ) and and nozzle (305); at the nozzle

The (305) has a cross-linking chamber (304) and a central shaft (303). The A-component syringe (301) and the B-component syringe (302) are connected to the cross-linking chamber (304).

7. A special device for preparing a medical biological tissue structure according to claim 5, wherein: said second The functional layer nozzle (102) and the protective film nozzle (101) are pneumatic spray valves, and the structure thereof comprises an internal pipe (602) and an external sleeve (604), and the pneumatic spray valve is connected to the high pressure gas source (111) through the gas pipeline. connection.

8. The special device for preparing a medical biological tissue structure according to claim 5, wherein: the first functional layer nozzle (103) comprises a rack (401), a solution storage box (402), and a screening a device (403) and a piezoelectric injection valve (404); a solution storage box (402), a screening device (403), and a piezoelectric injection valve (404) are mounted on the hanger (401);

(403) is installed under the solution storage box (402) and communicated through the pipeline; the screening device (403) includes a screening plate (501) and a substrate (502), and the substrate (502) is provided with a screening groove on the side ( 503), a screening plate (501) is installed on both sides of the front side of the screening tank (503).

A method of preparing a medical biological tissue structure according to claim 1, wherein the method comprises the following steps:

1) Under the control of the control unit (113), the X-direction moving device (105) and the Y-direction moving device (107) drive the rotary forming table (106) to move to a set position below the hollow tube forming nozzle (110). Starting a hollow tube forming nozzle (110), the natural polymer solution and the crosslinking agent in the hollow tube forming nozzle (110) are mixed and extruded to form one or a set of hollow tube structures arranged in parallel;

2) The rotary forming table (106) is moved by the X-direction moving device (105) and the Y-direction moving device (107) to move to the set position below the curved track (104) to activate the first functional layer nozzle (103) Spraying the biological material with or without cells onto the surface of the hollow tube; then starting the second functional layer nozzle (102), spraying the second or no cell-containing biological material onto the surface of the hollow tube, in the step 1) The formed hollow tube (801) structured surface forms a functional layer (802) containing multiple layers of biological material and cells;

3) The Z-direction moving device (107) drives the hollow tube forming nozzle (110) to move, and presses another parallel-arranged hollow tube (801) in the vertical direction of the parallel hollow tube formed in step 1), repeating step 2 Forming a composite formed body;

5) The protective film nozzle (101) is activated, and the synthetic polymer solution is sprayed in a mist form to form a protective film (803) on the surface of the composite molded body to produce a tissue organ precursor having a spatially complex shape and a composite material. .

The method for preparing a medical biological tissue structure according to claim 9, wherein: the natural polymer solution and the crosslinking agent in step 1) are sodium alginate solution and calcium chloride solution, respectively, or They are fibrinogen and thrombin solution, respectively, or collagen solution and cell culture medium solution, respectively, or polylactic acid glycolic acid solution and water, respectively, or polyurethane solution and water, respectively, or dicalcium phosphate dihydrate and hydrogen, respectively. Calcium oxide solution, or calcium dihydrogen phosphate monohydrate and calcium oxide solution, respectively, or tetracalcium phosphate and calcium hydrogen phosphate solution, or calcium phosphate aqueous solution and polylactic acid glycolic acid solution, respectively, or hydroxyapatite Water and P polylactic acid glycolic acid solution; wherein, sodium alginate solution and calcium chloride solution The concentration is: Dissolve sodium alginate in the cell culture medium at a concentration of 0.1-5. /. (w/v), the calcium chloride aqueous solution is in deionized water at a concentration of 1-10% (w/v); the concentration of fibrinogen and thrombin solution is: Dissolving fibrinogen powder in cell culture medium In the solution, the mass concentration is 0.01-5. /. (w/v), dissolving thrombin in deionized water at a concentration of 50-200 U rrL; concentration of collagen solution: dissolving collagen in cell culture medium at a concentration of 0.01-5%

(w/v) _{; the} concentration of calcium hydrogen phosphate dihydrate and calcium hydroxide solution are respectively: mixing calcium hydrogen phosphate and calcium hydroxide in a ratio of 1-20% (w/v) of mass concentration with 1M disodium hydrogen phosphate solution; The concentrations of calcium dihydrogen phosphate monohydrate and calcium oxide solution are as follows: The monohydrate calcium dihydrogen phosphate and calcium oxide particles are mixed with 1 M phosphate buffer at a mass concentration of 1 - 20% (w/v); The mass concentration of calcium and calcium hydrogen phosphate solution is: 1-20 for tetracalcium phosphate and calcium hydrogen phosphate. /. (w/v) mixed with phosphate buffer separately; polylactic acid glycolic acid solution and polyurethane solution concentration are: Polylactic acid glycolic acid, polyurethane dissolved in tetraethylene glycol, mass concentration of 0.1-15% (w / v _); step 2) comprises the biological material is a cell, growth factors, cell freezing medium, anticoagulant materials, drugs, gelatin, hyaluronic acid, collagen, fibroin, laminin, elastin, monosaccharides , one or more combinations of disaccharide, dextrose, mucopolysaccharide, heparin, chitosan, phosphorylated chitosan, sulfated chitosan and matrigel, wherein the cell density is ¹ ×10 ² / ΓΤ 1_- 1Χ10 ⁷ /rrl_, the cells are one or a combination of adult cells such as osteoblasts, hepatocytes, cardiomyocytes, stellate cells, fibroblasts, embryonic stem cells, induced pluripotent stem cells, and adipose stem cells. The growth factor concentration is 10-50 ng/rrL, and the cell cryopreservation volume is dimethyl sulfoxide aqueous solution, or glycerin solution, or dextrose solution, and the mass concentration is 1-20. 3⁄4^, anticoagulant material, drug, gelatin, hyaluronic acid, collagen, silk fibroin, laminin, elastin, monosaccharide, disaccharide, dextrose, mucopolysaccharide, heparin, chitosan, phosphorylation The mass concentration of chitosan, sulfated chitosan and matrigel is 0.1-20. 3⁄4w/v) _; The synthetic polymer solution described in the step 3) dissolves the synthetic polymer in an organic solvent, and the synthetic polymer is polyurethane, or polylactic acid glycolic acid, or polyethylene, or polypropylene, or poly One or a combination of caprolactone, or polycarbonate, or polyethylene glycol, or polyhydroxy acid ester, the organic solvent is synthesized using dimethyl sulfoxide, tetraethylene glycol or 1.4 dioxane The mass concentration of the polymer solution is 1-30% (w/v).