CN116328029A - Micro-channel photocrosslinking hydrogel tissue engineering scaffold formed based on electrostatic spinning and preparation method and application thereof - Google Patents
Micro-channel photocrosslinking hydrogel tissue engineering scaffold formed based on electrostatic spinning and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the field of tissue engineering and biomedical materials, in particular to a micro-pore photocrosslinked hydrogel tissue engineering scaffold formed based on electrostatic spinning, and a preparation method and application thereof. The micro-pore photo-crosslinking hydrogel tissue engineering scaffold consists of an inner layer membrane and a photo-crosslinking hydrogel shell, wherein the inner layer is a polyvinyl alcohol electrospun fiber membrane as a sacrificial material, and the shell is gelatin methacryloyl ultraviolet crosslinking hydrogel as a supporting material; the micro-pore channel photocrosslinked hydrogel tissue engineering scaffold can effectively form micro-pore channels in the photocrosslinked hydrogel scaffold, and can furthest improve the degree of new blood vessels in engineering tissues according to the theory of contact guiding, and can uniformly distribute oxygen and other nutrient substances in the scaffold with a regenerated large tissue structure, thereby having the functions of promoting adhesion, proliferation and migration of endothelial cells. The invention can be applied to the fields of engineering tissue vascularization and skin grafting biomedical materials.
Description
Technical Field
The invention relates to the field of tissue engineering and biomedical materials, in particular to a micro-pore photocrosslinked hydrogel tissue engineering scaffold formed based on electrostatic spinning, and a preparation method and application thereof.
Background
Induction of angiogenesis by humans at the site of injury has been a long-felt problem, and the concept that structures can guide the desired biological function has been applied to the physical design of three-dimensional channel networks at the site of implantation. (Panel-Tala S, macNeil S, claystens F. The tissue-engineered vascular graft-past, present, and future. Tissue Engineering Part B-Reviews, 2016, 22 (1): 68-100). Damaged or injured blood vessels can occur at various levels of the vascular system, from large blood vessels (e.g., arteries and veins) to medium and micro blood vessels (e.g., arterioles, venules, and capillary networks). Vascular tissue engineering has become a promising approach to construct small caliber vascular grafts for occlusion arteries. The goal of vascularized tissue engineering is to construct medium, microvasculature for the pre-vascularization of engineered tissues and organs. The ideal small diameter vascular graft has biocompatibility, bridging and mechanical stability, while maintaining patency, promoting tissue remodeling. After implantation, the desired medium and microvasculature should rapidly bind to the host vessel and allow for nutrient and waste exchange throughout the structure. (Wang Z, mithieux S M, weiss A S. Fabrication techniques for vascular and vascularized tissue engineering [ J ]. Advanced Healthcare Materials, 2019, 8 (19): 1900742.). The diameter of the vascular graft passageway is currently typically on the millimeter or sub-millimeter level, whereas the diameter of human capillaries is approximately 5-10 microns. (Zhou Y, sooriyaarachchi D, tan G Z Fabrication of nanopores polylactic acid microtubes by core-sheath electrospinning for capillary vascularization [ J ]. Biomimetics, 2021, 6 (1): 15.). However, in the initial stage of implantation of a common artificial vascular tissue engineering scaffold, cells are difficult to survive due to lack of oxygen, nutrients and the like, and adhesion, activation, accumulation and the like of platelets are easily initiated after blood is directly contacted with biological materials, so that the scaffold is unfavorable for regeneration of blood vessels.
Electrospinning offers the possibility of manufacturing a variety of ECM-like scaffolds with high surface area and porosity by using natural and synthetic polymers. It is well known that the process conditions applied by an electrospinning machine may affect the characteristics of the finally produced electrospun fibers (MAT). These conditions can be generalized to a specific group including the electrospinning environment (e.g., humidity and temperature); electrospinning parameters (e.g., applied voltage (in kV range), tip-to-collector distance (cm), and flow rate of solution (μl/min)); and properties of the polymer solution such as polymer molecular weight, solution viscosity and solvent volatility. For example, the fiber diameter is increased not only because of the increased concentration and molecular weight of the polymer used but also because of the increased solution flow rate. (Nazarnezhad S, baino F, kim H W, et al Electrospun nanofibers for improved angiogenesis: promises for tissue engineering applications [ J ]. Nanominates, 2020, 10 (8): 1609.).
Hydrogels mimic the extracellular matrix (ECM) surrounding cells and have been widely used in the fields of tissue engineering and regenerative medicine. Their high water content, tunable chemical and physical properties, and the ability to encapsulate cells, biological macromolecules (e.g., polypeptides/proteins, nucleotides, antibodies) and therapeutic agents opens up a variety of potential applications. Photoinduced photocrosslinking has been widely used to prepare cell-loaded hydrogels with rapid gel times and adjustable physical properties.
Disclosure of Invention
The invention overcomes the defects of the prior art and provides a micro-channel photocrosslinking hydrogel tissue engineering scaffold based on electrostatic spinning;
in order to solve the technical problems, the invention adopts the following technical scheme: the micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning consists of an inner layer membrane and a photocrosslinked hydrogel shell, wherein the inner layer is a polyvinyl alcohol electrospun fiber membrane as a sacrificial material, and the shell is gelatin methacryloyl ultraviolet crosslinked hydrogel as a supporting material; the polyvinyl alcohol fiber membrane formed by electrostatic spinning is used as a sacrificial material, and the photocrosslinked hydrogel is filled on the fiber membrane to form the photocrosslinked hydrogel tissue engineering scaffold containing micro-pore channels.
Further, the alcoholysis degree of the polyvinyl alcohol of the inner layer fiber film is 78.5-81.5 mol percent, and the viscosity is 45.0-51.0 mPa.s; the gelatin methacryloyl ultraviolet crosslinking hydrogel can be rapidly photocrosslinked, the mechanical property can reach 100 kPa, and the crosslinking time is 15-30 s.
In addition, the invention also provides a preparation method of the crosslinked hydrogel tissue engineering scaffold, namely, firstly, forming a sacrificial material through electrostatic spinning, then preparing a photocrosslinking hydrogel pre-crosslinked solution to be filled in a fibrous membrane, and eluting after ultraviolet crosslinking gel formation to form the hydrogel tissue engineering scaffold with micro-pore channels inside.
The invention takes polyvinyl alcohol and gelatin methyl acryloyl as raw materials, and prepares a micro-channel photo-crosslinking hydrogel tissue engineering scaffold material by a solution electrospinning and ultraviolet crosslinking gel forming method. The hydrogel tissue engineering scaffold material takes PVA fiber formed by electrostatic spinning as a sacrificial material, takes photo-crosslinked hydrogel GelMA as a filling material, can effectively form micro-channels in the photo-crosslinked hydrogel scaffold, improves the degree of new blood vessels in engineering tissues to the maximum extent according to the theory of contact guiding, can uniformly distribute oxygen and other nutrient substances in the scaffold with a regenerated large tissue structure, and has the functions of promoting endothelial cell adhesion, proliferation and migration.
Further, the preparation method specifically comprises the following steps:
(1) Dissolving polyvinyl alcohol in a mixed solvent of water and hexafluoroisopropanol;
(2) Electrospinning the solution obtained in the step (1) to obtain PVA microfibers;
(3) Adding PBS into a brown bottle filled with an initiator LAP, heating in a water bath at 40-50 ℃ for about 15 min under the condition of avoiding light, oscillating for several times until the solution is completely dissolved to obtain an initiator standard solution, adding the initiator standard solution into GelMA, oscillating to fully infiltrate the GelMA, heating in a water bath at 60-70 ℃ for avoiding light for 20-30 min, oscillating for several times until the solution is completely dissolved, and preparing into GelMA solution to be crosslinked;
(4) Injecting the GelMA pre-polymerization solution in the step (3) into a PDMS mold filled with the electrospun PVA microfiber in the step (2) by using a needle, and carrying out ultraviolet light source irradiation to gel the GelMA pre-polymerization solution;
(5) Soaking the photo-crosslinked GelMA hydrogel embedded with the PVA fibers obtained in the step (4) in PBS solution, continuously replacing the PBS solution until the PVA is completely dissolved in water, and then continuously flushing with PBS until the PVA is completely eluted, thereby obtaining the photo-crosslinked hydrogel tissue engineering scaffold.
Further, in the step (1), the product ratio of water to hexafluoroisopropanol is 1:2-4, and the concentration of the prepared polyvinyl alcohol solution is 100-150 mg/mL; when in dissolution, firstly, polyvinyl alcohol is dissolved in hexafluoroisopropanol, stirred for 6 hours, water is added after the polyvinyl alcohol is uniform, and stirring is continued to prepare uniform solution.
Further, the electrospinning conditions in the step (2) are as follows: the injection rate is 4-20 mL/h, the voltage of electrospinning is 5-15 kV, the receiving distance is 10-22 cm, and the electrospinning is performed for 0.2-0.5 h to obtain the electrospinning fiber material.
Further, the volume to mass ratio of PBS to initiator LAP is 5 ml:0.025 g; the mass volume ratio of GelMA to the initiator standard solution is 1000 mg:5 ml, the concentration of the resulting GelMA solution was 20 wt%.
Further, the needle used in the step (4) is a 27G needle; in the step (4), a 405 ultraviolet light source nm is used for irradiating 30 s to gel the GelMA prepolymerization solution, and the gel strength can be regulated and controlled through the irradiation time and the intensity; the photo-crosslinked GelMA hydrogel with PVA fibers embedded in step (5) had a soak time of 1 h in PBS solution.
In addition, the invention also provides application of the photocrosslinked hydrogel tissue engineering scaffold in tissue engineering capillary regeneration materials and application of the photocrosslinked hydrogel tissue engineering scaffold in skin grafting biomedical materials.
Compared with the prior art, the invention has the following beneficial effects:
the invention has the advantages that the photocrosslinking hydrogel tissue engineering scaffold material is internally provided with the micro-pore canal formed by the electrostatic spinning PVA fiber, and the problem of slow vascularization in vitro tissue culture can be effectively solved through contact guidance. The polyvinyl alcohol electrospinning sacrificial material is dissolved at room temperature, and when a micro-pore channel network is formed in the hydrogel tissue engineering scaffold as a supporting material, the polyvinyl alcohol electrospinning sacrificial material can be dissolved without heating, so that the requirements and the damage to the scaffold are reduced, and the polyvinyl alcohol electrospinning sacrificial material can play a role in temporary supporting in the pre-vascularized hydrogel tissue engineering scaffold. Meanwhile, antibacterial factors (such as ZnO, nano Ag and the like) and growth factors can be added into PVA to dissolve PVA so as to diffuse and release PVA, and promote migration, adhesion and proliferation of endothelial cells. The photocrosslinked hydrogel bracket material can be quickly glued in a short time (15-30 s), and can be quickly packaged by being filled into the sacrificial material, so that the PVA electrospinning sacrificial material is prevented from being dissolved in water and difficult to form micro-tunnels. In addition, the introduction of the micro-channels can effectively promote the migration of cells, thereby leading the cells in the hydrogel scaffold to become vascularized. Meanwhile, the GelMA photocrosslinking hydrogel stent material has better biocompatibility, is beneficial to the growth of endothelial cells into the micro-channel, promotes the formation of vascular buds at the initial stage of vascular formation, and can provide mechanical properties for vascular stents more similar to those of natural blood vessels. The invention is applied to the fields of tissue engineering capillary regenerated materials and skin grafting biomedical materials.
Drawings
FIG. 1 is an SEM photograph of a PVA electrospun film of about 10 μm prepared according to example 1 of the present invention.
FIG. 2 is a graph showing dissolution of 1. 1 h in PBS solution prior to preparation of a PVA electrospun film of about 10 μm according to example 1 of the present invention.
FIG. 3 is a photograph of a photo-crosslinked hydrogel scaffold prepared in example 1 of the present invention having anti-diffusion dye infused micro-tunnels.
FIG. 4 is an optical micrograph of a photo-crosslinked hydrogel scaffold prepared in example 1 of the present invention after being infused.
Detailed Description
The invention is further illustrated below with reference to specific examples.
The micro-channel photocrosslinked hydrogel tissue engineering scaffold formed by electrostatic spinning consists of an inner membrane and an outer shell, wherein the inner layer is a polyvinyl alcohol electrospun fiber membrane which is used as a sacrificial material and consists of fibers with the thickness of 500-1000 mu m and 1-45 mu m. The shell is gelatin methacryloyl ultraviolet crosslinked hydrogel, and the thickness of the gelatin methacryloyl ultraviolet crosslinked hydrogel is 1-2 mm as a supporting material.
The alcoholysis degree of the polyvinyl alcohol is 78.5-81.5 mol percent, and the viscosity is 45.0-51.0 mPa.s.
The preparation method of the micro-channel photocrosslinked hydrogel tissue engineering scaffold formed by electrostatic spinning comprises the following steps:
(1) And (3) dissolving polyvinyl alcohol in a mixed solution of water and hexafluoroisopropanol, wherein the volume ratio of the solvent is 1 (2-4), the concentration of the prepared solution is 100-150 mg/mL, dissolving the polyvinyl alcohol in hexafluoroisopropanol firstly during dissolving, stirring for 6h, adding water after being uniform, and continuously stirring to prepare a uniform solution.
(2) Electrospinning the solution obtained in the step (1), wherein the electrospinning conditions are as follows: the injection rate is 4-20 mL/h, the voltage of electrospinning is 5-15 kV, the receiving distance is 10-22 cm, and the electrospinning is performed for 0.2-0.5 h to obtain the electrospinning fiber material. Afterwards, the micro-fiber materials with different diameters are electrospun under the adjustment condition.
(3) 5 ml of PBS solution is taken and added into a brown bottle (containing 0.025 g of LAP) with the pink color of the initiator LAP, and the mixture is heated for about 15 min in a water bath at 40-50 ℃ under the condition of light shielding, and the mixture is oscillated for a plurality of times until the mixture is completely dissolved. Weighing 1000 mg GelMA, placing into a centrifuge tube, adding 5 ml initiator standard solution into the centrifuge tube, oscillating to fully infiltrate the GelMA, heating in a water bath at 60-70deg.C in the dark for 20-30 min, oscillating for several times until completely dissolving, preparing into GelMA solution with concentration of 20 wt%, and waiting for crosslinking.
(4) Injecting the GelMA pre-polymerization solution in the step (3) into a PDMS mold provided with the electrostatic spinning PVA microfiber in the step (2) by using a 27G needle, irradiating 15 s by using a 405 nm ultraviolet light source to gel the GelMA pre-polymerization solution, and regulating the gel strength by using the illumination time and the strength.
(5) Soaking the photo-crosslinked GelMA hydrogel embedded with the PVA fibers obtained in the step (4) in PBS solution for 1 h, continuously replacing the PBS solution until the PVA is completely dissolved in water, and then continuously flushing with PBS until the PVA is completely eluted.
Example 1
Polyvinyl alcohol is dissolved in a mixed solution of water and hexafluoroisopropanol, and the solvent volume ratio is 1:3, so that the solution with the concentration of 150mg/mL is prepared. When in dissolution, firstly, polyvinyl alcohol is dissolved in hexafluoroisopropanol, and then is stirred for 6h, and after the polyvinyl alcohol is dissolved uniformly, water is added, and stirring is continued to prepare uniform solution.
And (3) electrospinning the solution at the temperature of 31 ℃, the humidity of 57%, the injection rate of 7.2 mL/h, the electrospinning voltage of 15kV and the receiving distance of 17 cm, and electrospinning 0.25 h to obtain the electrospun fiber material with the thickness of 1 mm, wherein the electrospun fiber material consists of fibers with the diameter of 6.7-11.8 mu m.
The photoinitiator LAP was dissolved in PBS solution to prepare a 0.25. 0.25wt% LAP photoinitiator standard solution. Weighing 1000 mg GelMA, placing into a centrifuge tube, adding 5 ml initiator standard solution into the centrifuge tube, oscillating to fully infiltrate the GelMA, heating in a water bath at 65deg.C for 25 min in the dark, oscillating for several times until completely dissolving, and preparing into GelMA pre-crosslinked solution with concentration of 20 wt%.
Injecting the GelMA pre-crosslinked solution into a PDMS mold containing a PVA electro-spinning sacrificial material by using a 27G needle to fill the GelMA into the gaps of PVA fibers, immediately irradiating with 405 nm ultraviolet light source and 30 s to gel the GelMA pre-crosslinked solution, and regulating the gel strength by the irradiation time and the intensity. The resulting photo-crosslinked GelMA hydrogel embedded PVA fibers were soaked with PBS solution for 1 h, the PBS solution was continuously changed until PVA was completely dissolved in water, and then washed with PBS until complete elution.
SEM photographs of the electrospun PVA fibers are shown in fig. 1. The in-water degradation curve of the electrospun PVA fibers is shown in fig. 2. An optical micrograph of the micro-tunnels formed in the photo-crosslinked hydrogel scaffold is shown in fig. 3.
Example 2
Polyvinyl alcohol is dissolved in a mixed solution of water and hexafluoroisopropanol, and the volume ratio of the solvent is 1:4, so that the solution with the concentration of 100 mg/mL is prepared. When in dissolution, firstly, polyvinyl alcohol is dissolved in hexafluoroisopropanol, and then is stirred for 6h, and after the polyvinyl alcohol is dissolved uniformly, water is added, and stirring is continued to prepare uniform solution.
And (3) electrospinning the solution at the temperature of 32 ℃, the humidity of 50%, the injection rate of 3.6 mL/h, the voltage of electrospinning of 5kV and the receiving distance of 10 cm, and electrospinning 0.4 h to obtain the electrospun fiber material with the thickness of 1 mm, wherein the electrospun fiber material consists of fibers with the diameter of 7.2-19.4 mu m.
The photoinitiator LAP was dissolved in PBS solution to prepare a 0.25. 0.25wt% LAP photoinitiator standard solution. Weighing 1000 mg GelMA, placing into a centrifuge tube, adding 5 ml initiator standard solution into the centrifuge tube, oscillating to fully infiltrate the GelMA, heating in a water bath at 65deg.C for 25 min in the dark, oscillating for several times until completely dissolving, and preparing into 15 wt% GelMA pre-crosslinked solution.
Injecting the GelMA pre-crosslinked solution into a PDMS mold containing a PVA electro-spinning sacrificial material by using a 27G needle to fill the GelMA into the gaps of PVA fibers, immediately irradiating 20 s by using a 405 nm ultraviolet light source to gel the GelMA pre-crosslinked solution, and regulating the gel strength by using the irradiation time and the strength. The resulting photo-crosslinked GelMA hydrogel embedded PVA fibers were soaked with PBS solution for 1 h, the PBS solution was continuously changed until PVA was completely dissolved in water, and then washed with PBS until complete elution.
Example 3
Polyvinyl alcohol is dissolved in a mixed solution of water and hexafluoroisopropanol, and the volume ratio of the solvent is 1:3, so that the solution with the concentration of 100 mg/mL is prepared. When in dissolution, firstly, polyvinyl alcohol is dissolved in hexafluoroisopropanol, and then is stirred for 6h, and after the polyvinyl alcohol is dissolved uniformly, water is added, and stirring is continued to prepare uniform solution.
And (3) electrospinning the solution at the temperature of 32 ℃, the humidity of 45%, the injection rate of 18 mL/h, the voltage of 10 kV, the receiving distance of 12 cm and 0.2 h to obtain the electrospun fiber material with the thickness of 1 mm, wherein the electrospun fiber material consists of fibers with the diameter of 7.4-20.2 mu m.
The photoinitiator LAP was dissolved in PBS to prepare a 0.25wt% LAP photoinitiator standard solution. Weighing 1000 mg GelMA, placing into a centrifuge tube, adding 5 ml initiator standard solution into the centrifuge tube, oscillating to fully infiltrate the GelMA, heating in a water bath at 65deg.C for 25 min in the dark, oscillating for several times until completely dissolving, and preparing into GelMA pre-crosslinked solution with concentration of 20 wt%.
Injecting the GelMA pre-crosslinked solution into a PDMS mold containing a PVA electro-spinning sacrificial material by using a 27G needle to fill the GelMA into the gaps of PVA fibers, immediately irradiating 20 s by using a 405 nm ultraviolet light source to gel the GelMA pre-crosslinked solution, and regulating the gel strength by using the irradiation time and the strength. The resulting photo-crosslinked GelMA hydrogel embedded PVA fibers were soaked with PBS solution for 1 h, the PBS solution was continuously changed until PVA was completely dissolved in water, and then washed with PBS until complete elution.
Example 4
Polyvinyl alcohol is dissolved in a mixed solution of water and hexafluoroisopropanol, and the solvent volume ratio is 1:2, so that the solution with the concentration of 150mg/mL is prepared. When in dissolution, firstly, polyvinyl alcohol is dissolved in hexafluoroisopropanol, and then is stirred for 6h, and after the polyvinyl alcohol is dissolved uniformly, water is added, and stirring is continued to prepare uniform solution.
And (3) electrospinning the solution at the temperature of 30 ℃, the humidity of 59%, the injection rate of 14.4 mL/h, the voltage of 10 kV, the receiving distance of 17 cm and 0.2 h to obtain the electrospun fiber material with the thickness of 1 mm, wherein the electrospun fiber material consists of fibers with the diameter of 12.3-35.8 mu m.
The photoinitiator LAP was dissolved in PBS to prepare a 0.25wt% LAP photoinitiator standard solution. Weighing 1000 mg GelMA, placing into a centrifuge tube, adding 5 ml initiator standard solution into the centrifuge tube, oscillating to fully infiltrate the GelMA, heating in a water bath at 65deg.C for 25 min in the dark, oscillating for several times until completely dissolving, and preparing into GelMA pre-crosslinked solution with concentration of 20 wt%.
Injecting the GelMA pre-crosslinked solution into a PDMS mold containing a PVA electro-spinning sacrificial material by using a 27G needle to fill the GelMA into the gaps of PVA fibers, immediately irradiating with 405 nm ultraviolet light source and 30 s to gel the GelMA pre-crosslinked solution, and regulating the gel strength by the irradiation time and the intensity. The resulting photo-crosslinked GelMA hydrogel embedded PVA fibers were soaked with PBS solution for 1 h, the PBS solution was continuously changed until PVA was completely dissolved in water, and then washed with PBS until complete elution.
Claims (10)
1. The micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning is characterized by comprising an inner layer membrane and a photocrosslinked hydrogel shell, wherein the inner layer is a polyvinyl alcohol electrospun fiber membrane serving as a sacrificial material, and the shell is gelatin methacryloyl ultraviolet crosslinked hydrogel serving as a supporting material; the polyvinyl alcohol fiber membrane formed by electrostatic spinning is used as a sacrificial material, and the photocrosslinked hydrogel is filled on the fiber membrane to form the photocrosslinked hydrogel tissue engineering scaffold containing micro-pore channels.
2. The micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning according to claim 1, wherein the alcoholysis degree of the polyvinyl alcohol of the inner layer fiber film is 78.5-81.5 mol%, and the viscosity is 45.0-51.0 mpa.s; the gelatin methacryloyl ultraviolet crosslinking hydrogel can be rapidly photocrosslinked, the mechanical property can reach 100 kPa, and the crosslinking time is 15-30 s.
3. A preparation method of a micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning is characterized in that a sacrificial material is formed by electrostatic spinning, a photocrosslinked hydrogel precrosslinked solution is prepared and filled in a fibrous membrane, ultraviolet crosslinking is performed to gel, and then elution is performed to form the hydrogel tissue engineering scaffold containing micro-channels.
4. The method for preparing the micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning according to claim 3, which is characterized by comprising the following steps:
(1) Dissolving polyvinyl alcohol in a mixed solvent of water and hexafluoroisopropanol;
(2) Electrospinning the solution obtained in the step (1) to obtain PVA microfibers;
(3) Adding PBS into a brown bottle filled with an initiator LAP, heating in a water bath at 40-50 ℃ for about 15 min under the condition of avoiding light, oscillating for several times until the solution is completely dissolved to obtain an initiator standard solution, adding the initiator standard solution into GelMA, oscillating to fully infiltrate the GelMA, heating in a water bath at 60-70 ℃ for avoiding light for 20-30 min, oscillating for several times until the solution is completely dissolved, and preparing into the GelMA solution to be crosslinked;
(4) Injecting the GelMA pre-polymerization solution in the step (3) into a PDMS mold filled with the electrospun PVA microfiber in the step (2) by using a needle, and carrying out ultraviolet light source irradiation to gel the GelMA pre-polymerization solution;
(5) Soaking the photo-crosslinked GelMA hydrogel embedded with the PVA fibers obtained in the step (4) in PBS solution, continuously replacing the PBS solution until the PVA is completely dissolved in water, and then continuously flushing with PBS until the PVA is completely eluted, thereby obtaining the photo-crosslinked hydrogel tissue engineering scaffold.
5. The preparation method of the micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning is characterized in that the product ratio of water to hexafluoroisopropanol in the step (1) is 1:2-4, and the concentration of the prepared polyvinyl alcohol solution is 100-150 mg/mL; when in dissolution, firstly, polyvinyl alcohol is dissolved in hexafluoroisopropanol, stirred for 6 hours, water is added after the polyvinyl alcohol is uniform, and stirring is continued to prepare uniform solution.
6. The method for preparing a micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning according to claim 4, wherein the electrospinning conditions in the step (2) are as follows: the injection rate is 4-20 mL/h, the voltage of electrospinning is 5-15 kV, the receiving distance is 10-22 cm, and the electrospinning is performed for 0.2-0.5 h to obtain the electrospinning fiber material.
7. The method for preparing a micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning according to claim 4, wherein the volume-mass ratio of PBS to initiator LAP is 5 ml:0.025 g; the mass volume ratio of GelMA to the initiator standard solution is 1000 mg:5 ml, the concentration of the resulting GelMA solution was 20 wt%.
8. The method for preparing a micro-channel photocrosslinked hydrogel tissue engineering scaffold based on electrostatic spinning according to claim 4, wherein the needle used in the step (4) is a 27G needle; in the step (4), a 405 ultraviolet light source and a nm ultraviolet light source are used for irradiating 15 to 30 s to gel the GelMA prepolymerization solution; the photo-crosslinked GelMA hydrogel with PVA fibers embedded in step (5) had a soak time of 1 h in PBS solution.
9. Use of a photocrosslinked hydrogel tissue engineering scaffold according to any one of claims 1 to 2 or a photocrosslinked hydrogel tissue engineering scaffold obtained by the preparation method according to any one of claims 3 to 8 in tissue engineering capillary regeneration materials.
10. Use of the photocrosslinked hydrogel tissue engineering scaffold according to any one of claims 1 to 2 or the photocrosslinked hydrogel tissue engineering scaffold obtained by the preparation method according to any one of claims 3 to 8 in skin grafting biomedical materials.
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