CN116747353A - Preparation and application of photocrosslinked hydrogel based on Volton gel acellular matrix - Google Patents
Preparation and application of photocrosslinked hydrogel based on Volton gel acellular matrix Download PDFInfo
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- CN116747353A CN116747353A CN202310785181.2A CN202310785181A CN116747353A CN 116747353 A CN116747353 A CN 116747353A CN 202310785181 A CN202310785181 A CN 202310785181A CN 116747353 A CN116747353 A CN 116747353A
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- OARRHUQTFTUEOS-UHFFFAOYSA-N safranin Chemical compound [Cl-].C=12C=C(N)C(C)=CC2=NC2=CC(C)=C(N)C=C2[N+]=1C1=CC=CC=C1 OARRHUQTFTUEOS-UHFFFAOYSA-N 0.000 description 1
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
- A61L27/3633—Extracellular matrix [ECM]
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
- A61L27/3645—Connective tissue
- A61L27/3654—Cartilage, e.g. meniscus
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- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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Abstract
The application relates to the field of regenerative medicine, and discloses a preparation method of photocrosslinked hydrogel based on Voyton gum acellular matrix, which comprises the following steps of (1) taking umbilical cord tissue for acellular treatment to obtain acellular Voyton gum; wherein the umbilical cord tissue is derived from discarded human or animal umbilical cord; (2) The acellular wharton's jelly obtained by enzymatic digestion is utilized to obtain a water-soluble acellular matrix; (3) Dissolving a water-soluble acellular matrix in deionized water, and marking a photosensitive group to obtain a photosensitive acellular Waton gel material; (4) Preparing a photosensitive decellularized Walton gel material, adding a photoinitiator, and obtaining the photosensitive decellularized Walton gel matrix hydrogel material under the illumination of a light source. The acellular Wharton gum is obtained through acellular, enzymatic digestion and modification of a labeled photosensitive group, and then the acellular Wharton gum is mixed with a photoinitiator and cured through photo-crosslinking, so that the acellular umbilical cord Wharton gum matrix hydrogel is obtained, the preparation method is simple, the operability is strong, and the prepared hydrogel is high in mechanical strength and good in biocompatibility.
Description
Technical Field
The application relates to the technical field of regenerative medicine, in particular to preparation and application of photocrosslinked hydrogel based on a Volton gel decellularized matrix.
Background
Cartilage damage caused by trauma or osteoarthritis is a clinically common disease, causing inconvenience to the normal life of the patient. Repair of cartilage defects is also very challenging in clinical treatment. Tissue engineering provides a promising approach for cartilage regeneration and repair. As a core component of tissue engineering, scaffolds constructed from biological materials have a critical impact on cartilage regeneration. The good biological material has functional and structural complexity, can simulate the microenvironment provided by extracellular matrix, is beneficial to cell attachment and migration, nutrient diffusion, retention of growth factors and mechanical flexibility and adjustable mechanical rigidity.
To date, most scaffold materials for tissue engineering regeneration do not have the complexity of the natural extracellular matrix. Decellularized extracellular matrix-derived hydrogels have natural advantages in providing a tissue growth specific microenvironment, supporting cell growth and maturation, because they have the intrinsic components of the natural ECM, and all cellular components are removed by the decellularization process, avoiding adverse immune reactions.
The tissue engineering scaffold for repairing bones and cartilages is required to have good mechanical strength, and the mechanical strength of the acellular extracellular matrix is inevitably reduced in the preparation process, so that the problem that the acellular wharton's jelly needs to be solved in order to improve the mechanical strength of the acellular wharton's jelly can be used for cartilage tissue engineering.
Disclosure of Invention
Aiming at the defects of the prior art, the application provides a preparation method and application of photocrosslinked hydrogel based on a Walton gel acellular matrix, and solves the problem that the mechanical strength of the acellular extracellular matrix is reduced in the preparation process.
In order to achieve the above purpose, the application is realized by the following technical scheme: a preparation method of photocrosslinked hydrogel based on a Walton gel decellularized matrix comprises the following steps:
(1) Taking umbilical cord tissue, flushing with saline solution, removing adventitia and blood vessels, cutting into small pieces, and performing decellularization treatment to obtain decellularized Wayton gum; wherein the umbilical cord tissue is derived from discarded human or animal umbilical cord;
(2) The acellular wharton gel obtained by enzymatic digestion is centrifuged to remove supernatant, the reaction solution is dialyzed by deionized water, and the water-soluble acellular matrix is obtained after freeze drying;
(3) Dissolving the obtained water-soluble acellular matrix in deionized water, and marking the photosensitive group to obtain a photosensitive acellular Waton gel material; wherein, the marking method of the photosensitive group is to use acrylic anhydride molecules, methacrylic anhydride molecules, or acrylic glycidyl ester molecules, methacrylic glycidyl ester molecules, or acrylic chloride molecules, methacrylic chloride molecules;
(4) Preparing a photosensitive decellularized Walton gel material with the concentration of 5-20% w/v, adding a photoinitiator with the concentration of 0.1-1% w/v, and irradiating for 0.2-3min under the irradiation of light with the light source wavelength of 254-450nm to obtain the photosensitive decellularized Walton gel matrix hydrogel material.
Wherein, the labeling of the photosensitive group can be achieved by the following embodiments:
first embodiment: dissolving the decellularized Wauteron gum matrix in deionized water, cooling to 0-4 ℃, adding acrylic anhydride or methacrylic anhydride, slowly dripping 5M NaOH, reacting for 24 hours, pouring the reaction solution into a dialysis bag, dialyzing for 3-7d by using deionized water, and freeze-drying to obtain the photosensitive decellularized Wauteron gum matrix.
Second embodiment: dissolving a decellularized Wauteron gum matrix in deionized water, heating to 40 ℃ for stirring and dissolving, adding glycidyl acrylate or glycidyl methacrylate, adding 5M NaOH, reacting for 2-6h, pouring the reaction solution into a dialysis bag, dialyzing for 3-7d by using deionized water, and freeze-drying to obtain the photosensitive decellularized Wauteron gum matrix.
Third embodiment: dissolving a decellularized Wauteron gum matrix in anhydrous dimethyl sulfoxide, adding triethylamine, adding acryloyl chloride or methacryloyl chloride, dissolving in dichloromethane, reacting for 10 hours, pouring the reaction solution into ethanol for reprecipitation after the reaction is finished, dissolving the filtered crude product in deionized water again, dialyzing for 3-7d, and then freeze-drying to obtain the photosensitive decellularized Wauteron gum matrix.
Preferably, the salt solution in the step (1) is phosphate buffer solution.
Preferably, the decellularizing treatment in the step (1) includes:
s10, putting the Walton gel tissue into a 1M NaOH solution for decellularization for 3-6h;
s11, centrifuging to remove supernatant, placing the supernatant in a nuclease solution, dissolving the nuclease solution in a phosphate buffer solution, and stirring the solution for 2 to 8 hours at a constant temperature of 37 ℃;
s12, centrifuging to remove supernatant, placing in a phosphate buffer solution containing 5-15U/ml aprotinin, and oscillating at a constant temperature of 37 ℃ for 12-24h;
s13, centrifuging to remove the supernatant, and placing the supernatant in PBS solution for washing 3-5 times to obtain the decellularized Wauteron gel.
Preferably, in the step S11, the nuclease solution contains 30-70U/ml of DNase and 0.5-3U/ml of RNase A.
Preferably, in the step (2), the enzyme used in the enzymatic digestion is a biological enzyme capable of digesting collagen or polysaccharide macromolecules; the enzyme comprises collagenase, pepsin and pancreatin; the enzymatic digestion time is 6-48h.
Preferably, the photoinitiator is a water-soluble photoinitiator, preferably I2959, LAP, etc.
The application provides application of a photosensitive decellularized Walton gum matrix hydrogel material in the field of cartilage defect repair and regeneration, and particularly in the field of articular cartilage defect repair.
The application provides a preparation method and application of photocrosslinked hydrogel based on a Walton gel decellularized matrix. The beneficial effects are as follows:
1. according to the application, the acellular umbilical cord wharton gel matrix hydrogel is obtained through acellular, pepsin digestion and methacrylic anhydride modification, and is obtained through photo-crosslinking and solidification by mixing with a photoinitiator.
2. The raw material Wobbe gel for preparing the photocrosslinked hydrogel has wide sources, so that the photosensitive decellularized Wobbe gel has clinical feasibility for repairing cartilage defects, has the characteristics of injectability and rapid prototyping, can be used for preparing scaffolds with various forms by a 3D printing technology, and is beneficial to repairing cartilage defects.
Drawings
FIG. 1 is a photograph of the umbilical cord and, the Waton's gum removed from the adventitia and blood vessels of the umbilical cord, the general appearance of the Waton's gum after decellularization, histochemical staining and DAPI staining; wherein g is the cord tissue safranine solid-green staining, h is the cord tissue alisxin blue nucleus solid-red staining;
FIG. 2 is a nuclear magnetic hydrogen spectrum of a photosensitive decellularized Walton gum matrix;
FIG. 3 is a schematic diagram of a photosensitive decellularized Waton gum gel;
FIG. 4 is a freeze-dried electron microscope image of a photosensitive decellularized Waton gum;
FIG. 5 is a Live/read staining chart of photosensitive decellularized Walton gel loaded bone marrow mesenchymal stem cells.
Detailed Description
The following description of the embodiments of the present application will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
Embodiment one: preparation of photosensitive decellularized gum matrix
Preparation of a photosensitive decellularized Wauteron gum matrix: fresh umbilical cord tissue was taken, washed with phosphate buffer, adventitia and blood vessels were removed, and the umbilical cord tissue was cut into small pieces with scissors (as shown in fig. 1). Then the following steps are carried out in turn:
1) Immersing the clean Walton gum tissue into 1mol L-1NaOH solution for decellularization for 3 hours;
2) Centrifuging to remove supernatant, placing in nuclease solution (containing 50U/ml DNase and 1U/ml RNase A), dissolving in phosphate buffer solution, stirring at 37deg.C under constant temperature for 4 hr;
3) Centrifuging to remove supernatant, placing in phosphate buffer solution containing aprotinin 10U/ml, and oscillating at 37deg.C for 20 hr;
4) The supernatant was removed by centrifugation and washed well in PBS.
5) Finally, the acellular Waton gum matrix is sheared and placed in a refrigerator at the temperature of minus 80 ℃ for standby.
Preparation of a water-soluble acellular wharton's jelly matrix:
placing the acellular Waton gum matrix in 0.1% w/v pepsin solution (PH=2.0), vibrating and stirring at 37 ℃ for 24 hours, centrifuging to remove the supernatant, pouring the reaction solution into a dialysis bag (MWCO 3500), dialyzing for 3-7d with deionized water, and freeze-drying to obtain the water-soluble acellular matrix.
Preparation of methacryloyl acellular matrix:
dissolving 1g of water-soluble acellular matrix in deionized water, cooling to 0-4 ℃, adding 2mL of methacrylic anhydride, adjusting the pH to 8.5 with 5M NaOH, reacting for 24h, pouring the reaction solution into a dialysis bag (MWCO 3500), dialyzing for 3-7d with deionized water, and freeze-drying to obtain the methacryloyl acellular Wobton gum matrix.
The grafting of the decellularized Waton gum double bond was identified by nuclear magnetic resonance spectroscopy (as shown in FIG. 2).
Embodiment two: preparation of photosensitive decellularized Walton gum matrix hydrogel, gel formation general diagram and characterization of photosensitive decellularized matrix hydrogel preparation:
by formulating a concentration of the photosensitive decellularized matrix precursor solution (5% w/v-20% w/v), the hydrogel is rapidly crosslinked under light (405 nm) to form a hydrogel (as shown in FIG. 3).
Embodiment III: rheological and microstructure analysis of photosensitive decellularized wharton's jelly matrix hydrogels
By formulating a concentration of the photosensitive Waton gum decellularized matrix precursor solution (5% w/v-20% w/v), the hydrogel is formed by rapid crosslinking under light (405 nm). Freezing in-80deg.C refrigerator for 12 hr, lyophilizing, spraying gold, and observing microscopic morphology under scanning electron microscope (shown in figure 4).
Embodiment four: photosensitive decellularized wharton gel matrix hydrogel biocompatibility analysis
The rabbit bone marrow mesenchymal stem cells were sterilized by preparing a concentration of the photosensitive Waton's gel decellularized matrix precursor solution (5% w/v-20% w/v), mixing the rabbit bone marrow mesenchymal stem cells with the photosensitive Waton's gel decellularized matrix precursor solution uniformly at a cell amount of 2mil/ml, rapidly crosslinking to form a hydrogel under illumination (405 nm), culturing in a 24-well plate, performing Calcein AM/PI staining after 24 hours, and analyzing the biocompatibility (as shown in FIG. 5).
Although embodiments of the present application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the application, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. The preparation method of the photocrosslinked hydrogel based on the Walton gel acellular matrix is characterized by comprising the following steps of:
(1) Taking umbilical cord tissue, flushing with saline solution, removing adventitia and blood vessels, cutting into small pieces, and performing decellularization treatment to obtain decellularized Wayton gum;
(2) The acellular wharton gel obtained by enzymatic digestion is centrifuged to remove supernatant, the reaction solution is dialyzed by deionized water, and the water-soluble acellular matrix is obtained after freeze drying;
(3) Dissolving the obtained water-soluble acellular matrix in deionized water, and marking the photosensitive group to obtain a photosensitive acellular Waton gel material; wherein, the marking method of the photosensitive group is to use acrylic anhydride molecules, methacrylic anhydride molecules, or acrylic glycidyl ester molecules, methacrylic glycidyl ester molecules, or acrylic chloride molecules, methacrylic chloride molecules;
(4) Preparing a photosensitive decellularized Walton gel material with the concentration of 5-20% w/v, adding a photoinitiator with the concentration of 0.1-1% w/v, and irradiating for 0.2-3min under the irradiation of light with the light source wavelength of 254-450nm to obtain the photosensitive decellularized Walton gel matrix hydrogel material.
2. The method for preparing a photocrosslinked hydrogel based on a Waton gum cell matrix according to claim 1, wherein the salt solution in the step (1) is a phosphate buffer solution.
3. The method for preparing a photocrosslinked hydrogel based on a Waton gum decellularized matrix as claimed in claim 1, wherein the decellularizing treatment in the step (1) comprises:
s10, putting the Walton gel tissue into a 1M NaOH solution for decellularization for 3-6h;
s11, centrifuging to remove supernatant, placing the supernatant in a nuclease solution, dissolving the nuclease solution in a phosphate buffer solution, and stirring the solution for 2 to 8 hours at a constant temperature of 37 ℃;
s12, centrifuging to remove supernatant, placing in a phosphate buffer solution containing 5-15U/ml aprotinin, and oscillating at a constant temperature of 37 ℃ for 12-24h;
s13, centrifuging to remove the supernatant, and placing the supernatant in PBS solution for washing 3-5 times to obtain the decellularized Wauteron gel.
4. The method for preparing a photocrosslinked hydrogel based on a Waton' S cell matrix according to claim 3, wherein in the step S11, the nuclease solution contains 30-70U/ml of DNase and 0.5-3U/ml of RNase A.
5. The method for preparing a photocrosslinked hydrogel based on a Waton gum cell matrix according to claim 1, wherein in the step (2), the enzyme used for enzymatic digestion is a biological enzyme capable of digesting collagen or polysaccharide macromolecules; the enzyme comprises collagenase, pepsin and pancreatin; the enzymatic digestion time is 6-48h.
6. The method for preparing a photocrosslinked hydrogel based on a Waton gum acellular matrix according to claim 5, wherein in the step (3), when acrylic anhydride or methacrylic anhydride is used for marking a photosensitive group, the acellular Waton gum acetum is firstly dissolved in deionized water, cooled to 0-4 ℃, acrylic anhydride or methacrylic anhydride is added, 5M NaOH is slowly added dropwise for reaction for 24 hours, then the reaction solution is poured into a dialysis bag, dialyzed for 3-7d by deionized water, and then freeze-dried, so that the photosensitive acetum gum acetum matrix is obtained.
7. The method for preparing photocrosslinking hydrogel based on a Waton gum acellular matrix according to claim 1, wherein in the step (3), when the photosensitive group is marked by using glycidyl acrylate or glycidyl methacrylate, the acellular Waton gum matrix is firstly dissolved in deionized water, heated to 40 ℃ and stirred for dissolution, the glycidyl acrylate or the glycidyl methacrylate is added, then 5M NaOH is added, after the reaction is carried out for 2-6 hours, the reaction solution is poured into a dialysis bag, dialyzed for 3-7d by deionized water, and then freeze-dried, so that the photosensitive acellular Waton gum matrix is obtained.
8. The method for preparing photocrosslinking hydrogel based on a Volton gel acellular matrix according to claim 1, wherein in the step (3), when the photosensitive group is marked by using the acryloyl chloride or the methacryloyl chloride, the acellular Volton gel matrix is firstly dissolved in anhydrous dimethyl sulfoxide, triethylamine is added, then the acryloyl chloride or the methacryloyl chloride is added, the acellular Volton gel matrix is dissolved in dichloromethane, the reaction is carried out for 10 hours, after the reaction is finished, the reaction solution is poured into ethanol for reprecipitation, the crude product obtained by filtration is redissolved in deionized water, dialyzed for 3-7d, and then freeze-dried, so that the photosensitive acellular Volton gel matrix can be obtained.
9. The method for preparing a photocrosslinked hydrogel based on a Waton gum decellularized matrix as claimed in claim 1, wherein the photoinitiator is a water-soluble photoinitiator.
10. Use of a method for preparing a photocrosslinked hydrogel based on a wharton's jelly cell matrix according to any of claims 1 to 9 in the field of cartilage defect repair and regeneration.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102198292A (en) * | 2010-03-26 | 2011-09-28 | 卢世璧 | Scaffolds of umbilical cord decellularized Wharton jelly for tissue engineering and preparation method thereof |
| US20130095143A1 (en) * | 2010-03-30 | 2013-04-18 | Histocell, S.L. | Biomaterial from wharton's jelly umbilical cord |
| CN109675114A (en) * | 2018-12-20 | 2019-04-26 | 康泽生医学生物科技(武汉)有限公司 | A kind of preparation method of people's umbilical cord China Tong Shi glue tissue engineering bracket |
| CN115475279A (en) * | 2021-05-31 | 2022-12-16 | 上海交通大学医学院附属第九人民医院 | Photosensitive cartilage acellular matrix hydrogel material and preparation method and application thereof |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102198292A (en) * | 2010-03-26 | 2011-09-28 | 卢世璧 | Scaffolds of umbilical cord decellularized Wharton jelly for tissue engineering and preparation method thereof |
| US20130095143A1 (en) * | 2010-03-30 | 2013-04-18 | Histocell, S.L. | Biomaterial from wharton's jelly umbilical cord |
| CN109675114A (en) * | 2018-12-20 | 2019-04-26 | 康泽生医学生物科技(武汉)有限公司 | A kind of preparation method of people's umbilical cord China Tong Shi glue tissue engineering bracket |
| CN115475279A (en) * | 2021-05-31 | 2022-12-16 | 上海交通大学医学院附属第九人民医院 | Photosensitive cartilage acellular matrix hydrogel material and preparation method and application thereof |
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