CN106188442B - Chitosan derivative hydrogel and preparation method thereof - Google Patents
Chitosan derivative hydrogel and preparation method thereof Download PDFInfo
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- CN106188442B CN106188442B CN201610625357.8A CN201610625357A CN106188442B CN 106188442 B CN106188442 B CN 106188442B CN 201610625357 A CN201610625357 A CN 201610625357A CN 106188442 B CN106188442 B CN 106188442B
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- MQDJYUACMFCOFT-UHFFFAOYSA-N bis[2-(1-hydroxycyclohexyl)phenyl]methanone Chemical compound C=1C=CC=C(C(=O)C=2C(=CC=CC=2)C2(O)CCCCC2)C=1C1(O)CCCCC1 MQDJYUACMFCOFT-UHFFFAOYSA-N 0.000 claims description 4
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F299/00—Macromolecular compounds obtained by interreacting polymers involving only carbon-to-carbon unsaturated bond reactions, in the absence of non-macromolecular monomers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A—HUMAN NECESSITIES
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
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- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F2/00—Processes of polymerisation
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- C08F2/48—Polymerisation initiated by wave energy or particle radiation by ultraviolet or visible light
Abstract
The invention discloses a preparation method of chitosan derivative hydrogel, belonging to the field of biomedical materials. The preparation method of the chitosan derivative hydrogel comprises the steps of firstly preparing hydroxyethyl chitosan by a series of technological processes of alkalization, swelling, chlorohydrin modification, post-treatment and the like of chitosan, then carrying out graft modification by glycidyl methacrylate, and finally carrying out photochemical polymerization to form the hydrogel. The obtained hydrogel has the advantages of good biocompatibility, low toxicity, short gelling time, controllable and adjustable crosslinking density, good elasticity, good structural stability and the like. The preparation method has the characteristics of easily obtained raw materials, simple process, mild reaction conditions and the like, and the obtained hydrogel can be used for cell delivery, tissue engineering, three-dimensional cell culture and the like.
Description
Technical Field
The invention belongs to the field of biomedical materials, and particularly relates to a methacryloyl hydroxyethyl chitosan hydrogel and a preparation method thereof.
Background
The hydrogel is a polymer having a three-dimensional network structure, is generally formed by mutual crosslinking through covalent bonds, hydrogen bonds or van der waals forces, and absorbs a large amount of water in water to swell without dissolving. The hydrogel has various types, and has different classification methods based on different standards, and can be classified into natural polymer hydrogel and synthetic polymer hydrogel according to different material sources. The chitosan hydrogel is a typical representative of natural polymer hydrogel, has the advantages of low toxicity, good biocompatibility, biodegradability and the like, and is widely concerned in the field of biomedical materials. The chitosan which is the main raw material is derived from chitin, has the yield which is second to that of natural macromolecular polysaccharide of cellulose in nature, has wide sources, low price and easy obtainment and good biological performance, can be applied to the fields of environmental engineering, paper making, textile, food, medical treatment, biotechnology, pharmacy and the like, and particularly has wide application prospect in the research of biomedical materials, gene vectors and novel drug delivery systems. The chitosan molecular structure contains abundant active groups such as amino, hydroxyl and the like, and usually shows extremely strong affinity and modifiability. However, the solubility of chitosan in water is low, which greatly limits its application in biomedical materials. In previous reports, active hydroxyl and amino on a sugar ring in a chitosan molecular structure are chemically modified, so that the physical and chemical properties of chitosan are changed, the biological function of the chitosan is improved, and carboxymethylated chitosan is mainly used. Hydroxyethyl chitosan, as an important chitosan derivative, has better water solubility and biocompatibility, has higher solubility in organic solvents, reduces the crystallization property and hydrogen bonding effect of chitosan, and can improve the reaction activity [ CH Zhong, BHLuo, CR Zhou, et al.Synthesis of hydroxyl chitosan and its compatibility with poly (D, L-lactate) ]. chemical research and application,2010,22: 1102-.
The formation of carboxymethylated chitosan hydrogel is generally divided into chemical crosslinking and physical crosslinking, and the currently common methods are as follows: (1) crosslinking it using some chemical crosslinking agent; (2) the illumination or radiation and other macromolecules form a semi-interpenetrating or interpenetrating network; (3) some polyelectrolytes are added to cross-link them by electrostatic interaction or hydrogen bonding. The chemical crosslinking is usually carried out by using bifunctional crosslinking agents such as formaldehyde, glutaraldehyde and the like, so that the chitosan is crosslinked into gel through Schiff base, but the crosslinking agents have high toxicity, influence on cell activity and limit the research and application of the crosslinking agents in cells and organisms. Photo-crosslinking is a common way to construct hydrogels, and has mild reaction conditions, few byproducts, easily controlled reaction process, and high reaction efficiency, and is a good choice for constructing hydrogels [ XJYao, CX Pi, RM Li, et al. the Study on the synthesis of hydrogel by chemical Research,2002,13:22-24 ]. The hydrogel is prepared by utilizing a photo-crosslinking method, the curing process is relatively short, and the hydrogel can be gelled in situ. Therefore, the preparation of hydrogels by photocrosslinking has received much attention.
Disclosure of Invention
In order to overcome the defects of the existing chitosan derivative hydrogel and the preparation method thereof, the invention aims to provide a novel chitosan derivative hydrogel and the preparation method thereof. The hydrogel has the characteristics of injectability, short gelling time, adjustable crosslinking density, good elasticity, stable structure and the like. The preparation method has the advantages of easily obtained raw materials, simple process, mild reaction conditions and the like, and has wide application prospect in the aspects of cell delivery, tissue engineering, three-dimensional cell culture and the like.
In order to achieve the purpose, the invention adopts the following technical scheme.
A preparation method of chitosan derivative hydrogel comprises the steps of dissolving hydroxyethyl chitosan modified by glycidyl methacrylate in distilled water to form a solution with the mass concentration of 1% -4%, then adding a photoinitiator with the mass concentration of 0.05% -0.5% of the hydroxyethyl chitosan modified by glycidyl methacrylate, uniformly mixing, and irradiating by ultraviolet light for 0.5-8 minutes to obtain the hydroxyethyl chitosan hydrogel, namely the chitosan derivative hydrogel.
The invention further improves the preparation method of the hydroxyethyl chitosan modified by the glycidyl methacrylate, which comprises the following steps: preparing 0.2-2% hydroxyethyl chitosan solution, adding glycidyl methacrylate with 0.1-0.9 times of the number of repeating units of hydroxyethyl chitosan, adjusting the pH value to 8.0, magnetically stirring for 3-8 hours at 65-85 ℃, and dialyzing for 3 days by deionized water; and finally, freeze-drying the dialyzed product solution to obtain the glycidyl methacrylate modified hydroxyethyl chitosan.
A further development of the invention consists in that the pH is adjusted to 8.0 with a NaOH solution having a concentration of 1 mol/L.
The invention further improves that the cut-off molecular weight of the dialysis bag used for dialysis is 3500 Da.
The further improvement of the invention is that the freeze drying is carried out at minus 20 ℃ for 72 hours and at 20 ℃ for 4 hours.
The further improvement of the invention is that the preparation method of the hydroxyethyl chitosan comprises the following steps:
1) dispersing chitosan into a NaOH solution with the mass concentration of 30-60%, wherein the mass ratio of the chitosan to the NaOH is 3: 5; adding an isopropanol solution of chloroethanol, and then heating to 65 ℃ for reflux reaction for 4-7 hours; wherein, the number of the chitosan repeating units is as follows: chloroethanol: the molar ratio of the isopropanol is 1:3: 5-1: 6: 9;
2) after the reaction is finished, regulating the pH value to be neutral by hydrochloric acid, washing the obtained product for several times by using ethanol with the volume concentration of 75-90% and absolute ethanol, dissolving the product in water after vacuum drying, dialyzing for 48 hours, and then freeze-drying to obtain the hydroxyethyl chitosan.
The invention has the further improvement that the viscosity-average molecular mass of the chitosan is 300kDa to 1000kDa, and the deacetylation degree is more than 90 percent.
The further improvement of the invention is that the photoinitiator is 2-hydroxy-4' - (2-hydroxyethoxy) -2-methyl propiophenone or 1-hydroxycyclohexyl phenyl ketone.
The further improvement of the invention is that the parameters of the ultraviolet radiation are that the central wavelength is 365nm, and the light intensity is 1200mJ/cm2～1500mJ/cm2The irradiation distance was 15 cm.
A chitosan derivative hydrogel having a porous structure in communication with each other and having an average pore size of 165 μm.
Compared with the existing chitosan derivative hydrogel and the preparation technology, the invention has the following beneficial effects:
(1) the invention takes hydroxyethyl chitosan modified by glycidyl methacrylate as a raw material, adopts a photochemical polymerization mechanism to form the chitosan derivative hydrogel, and has the advantages of mild reaction conditions (room temperature), high reaction speed (completion within a few minutes), simple process and the like. And the whole gel preparation process takes water as a solvent, does not need to use toxic organic solvents, and has good safety performance.
(2) The hydrogel has short gelling time, can form gel in situ, can realize in-situ embedding and in-vivo injection of cells, can fill damaged parts in any shapes by delivering gel solution into specific parts in vivo, and achieves the aim of closely contacting materials with surrounding tissues. And does not need surgical operation, can reduce the pain of the patient.
(3) The hydrogel has a connected porous structure, uniform pore size distribution and an average pore size of about 165 μm. The hydrogel has the advantages of good elasticity, stable mechanical property and the like, and can be used for cell delivery, tissue engineering, three-dimensional cell culture and the like.
Furthermore, the main raw material of the hydrogel is chitosan, and the hydrogel has the advantages of easily obtained raw materials, low harm, good biocompatibility and the like. And the hydroxyethyl chitosan is prepared by using the chlorohydrin modified chitosan, so that the water solubility of the chitosan is well improved.
Drawings
FIG. 1 shows the preparation of glycidyl methacrylate-modified hydroxyethyl chitosan according to the present invention1HNMR spectra.
FIG. 2 shows the change of the modulus of hydroxyethyl chitosan hydrogel with the increase of irradiation time.
FIG. 3 shows the change of storage modulus (G ') and loss modulus (G') with increasing frequency of hydroxyethyl chitosan of the present invention.
FIG. 4 is a graph showing the swelling properties of hydroxyethyl chitosan hydrogels of the present invention.
FIG. 5 is a scanning electron micrograph of a hydroxyethyl chitosan hydrogel of the present invention, wherein FIG. 5(a) is at 35 times magnification and FIG. 5(b) is at 500 times magnification.
Detailed Description
The present invention is further described below by way of examples, but the present invention is not limited thereto.
Example 1
1) Dispersing chitosan with the viscosity-average molecular mass of 700kDa and the deacetylation degree of 98% in NaOH solution with the mass concentration of 50%, wherein the mass ratio of the chitosan to the NaOH in the NaOH solution is 3: 5; adding an isopropanol solution of chloroethanol, and then heating to 65 ℃ for reflux reaction for 5 hours; wherein, the number of the chitosan repeating units is as follows: chloroethanol: the molar ratio of isopropanol was 1:4.5: 7.
2) After the reaction is finished, regulating the pH value to be neutral by hydrochloric acid, washing the obtained product for several times by using ethanol with the volume concentration of 85% and absolute ethanol, dissolving the product in water after vacuum drying, dialyzing for 48 hours by using a dialysis bag with the molecular weight cutoff of 3500Da, and then freeze-drying, specifically keeping the product at minus 20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the hydroxyethyl chitosan.
3) Preparing a hydroxyethyl chitosan solution with the mass concentration of 1%, then adding glycidyl methacrylate with the repeating unit number of 0.4 time of that of the hydroxyethyl chitosan, adjusting the pH value to 8.0 by adopting a NaOH solution with the concentration of 1mol/L, magnetically stirring for 6 hours at 70 ℃, and dialyzing for 3 days by deionized water by adopting a dialysis bag with the molecular weight cutoff of 3500 Da; and finally, freeze-drying the dialysis product solution, specifically keeping the dialysis product solution at-20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the glycidyl methacrylate modified hydroxyethyl chitosan.
4) Condensing methacrylic acidDissolving the hydroxyethyl chitosan modified by the water glyceride into distilled water to form a solution with the mass concentration of 3.5%, then adding a photoinitiator with the mass of 0.1% of that of the hydroxyethyl chitosan modified by the glycidyl methacrylate, magnetically stirring and uniformly mixing, and then irradiating by using ultraviolet light for 2 minutes to obtain hydroxyethyl chitosan hydrogel, namely chitosan derivative hydrogel; wherein the photoinitiator is 2-hydroxy-4' - (2-hydroxyethoxy) -2-methyl propiophenone; the parameters of ultraviolet irradiation are as follows: the central wavelength is 365nm, and the light intensity is 1200mJ/cm2～1500mJ/cm2The irradiation distance was 15 cm.
As can be seen from FIG. 1, the chemical positions at 5.63ppm and 6.06ppm are the chemical shifts of adjacent protons on the carbon-carbon double bond in glycidyl methacrylate, confirming that glycidyl methacrylate-modified hydroxyethyl chitosan was successfully synthesized.
As can be seen from FIG. 2, the gel formation time of the hydrogel was 5.6 seconds, and the gel formation time was short.
As can be seen from FIG. 3, the storage modulus G 'is much greater than the loss modulus G', which indicates that the hydrogel has good elastic properties, and the modulus remains substantially unchanged with the change of frequency, so that the gel has strong structural stability.
As can be seen from FIG. 4, the hydrogel absorbed water very rapidly, had an equilibrium swelling time of about 12 hours and a maximum swelling ratio of 12, indicating that the hydrogel absorbed water rapidly and had a stable three-dimensional network structure.
FIG. 5 is a scanning electron microscope image of the hydrogel with 35 times and 500 times magnification, respectively, and from FIG. 5(a) and FIG. 5(b), it can be seen that the gel has a connected porous structure, which is favorable for the communication of water and nutrients, and has uniform pore size distribution of about 165 μm, such pore size is suitable for the growth of cells, and the pore walls are smooth.
Example 2
1) Dispersing chitosan with the viscosity-average molecular mass of 300kDa and the deacetylation degree of 95% in a NaOH solution with the mass concentration of 30%, wherein the mass ratio of the chitosan to the NaOH in the NaOH solution is 3: 5; adding an isopropanol solution of chloroethanol, and then heating to 65 ℃ for reflux reaction for 7 hours; wherein, the number of the chitosan repeating units is as follows: chloroethanol: the molar ratio of isopropanol was 1:3: 5.
2) After the reaction is finished, regulating the pH value to be neutral by hydrochloric acid, washing the obtained product for several times by using ethanol with the volume concentration of 85% and absolute ethanol, dissolving the product in water after vacuum drying, dialyzing for 48 hours by using a dialysis bag with the molecular weight cutoff of 3500Da, and then freeze-drying, specifically keeping the product at minus 20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the hydroxyethyl chitosan.
3) Preparing a hydroxyethyl chitosan solution with the mass concentration of 2%, then adding glycidyl methacrylate with the repeating unit number of 0.1 time of that of the hydroxyethyl chitosan, adjusting the pH value to 8.0 by adopting a NaOH solution with the concentration of 1mol/L, magnetically stirring for 6 hours at 70 ℃, and dialyzing for 3 days by deionized water by adopting a dialysis bag with the molecular weight cutoff of 3500 Da; and finally, freeze-drying the dialysis product solution, specifically keeping the dialysis product solution at-20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the glycidyl methacrylate modified hydroxyethyl chitosan.
4) Dissolving hydroxyethyl chitosan modified by glycidyl methacrylate in distilled water to form a solution with the mass concentration of 1%, then adding a photoinitiator with the mass of 0.2% of that of the hydroxyethyl chitosan modified by the glycidyl methacrylate, magnetically stirring and uniformly mixing, and then irradiating by using ultraviolet light for 0.5 minute to obtain hydroxyethyl chitosan hydrogel, namely chitosan derivative hydrogel; wherein the photoinitiator is 2-hydroxy-4' - (2-hydroxyethoxy) -2-methyl propiophenone; the parameters of ultraviolet irradiation are as follows: the central wavelength is 365nm, and the light intensity is 1200mJ/cm2The irradiation distance was 15 cm.
Example 3
1) Dispersing chitosan with the viscosity-average molecular mass of 1000kDa and the deacetylation degree of 91 percent in NaOH solution with the mass concentration of 60 percent, wherein the mass ratio of the chitosan to the NaOH in the NaOH solution is 3: 5; adding an isopropanol solution of chloroethanol, and then heating to 65 ℃ for reflux reaction for 4 hours; wherein, the number of the chitosan repeating units is as follows: chloroethanol: the molar ratio of isopropanol was 1:4: 7.
2) After the reaction is finished, regulating the pH value to be neutral by hydrochloric acid, washing the obtained product for several times by using 75% ethanol and absolute ethanol, dissolving the product in water after vacuum drying, dialyzing for 48 hours by using a dialysis bag with the molecular weight cutoff of 3500Da, and then freeze-drying, specifically keeping the product at minus 20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the hydroxyethyl chitosan.
3) Preparing a hydroxyethyl chitosan solution with the mass concentration of 1.5%, then adding glycidyl methacrylate with the repeating unit number of 0.4 time of that of the hydroxyethyl chitosan, adjusting the pH value to 8.0 by adopting a NaOH solution with the concentration of 1mol/L, magnetically stirring for 8 hours at 65 ℃, and dialyzing for 3 days by using deionized water through a dialysis bag with the molecular weight cutoff of 3500 Da; and finally, freeze-drying the dialysis product solution, specifically keeping the dialysis product solution at-20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the glycidyl methacrylate modified hydroxyethyl chitosan.
4) Dissolving hydroxyethyl chitosan modified by glycidyl methacrylate in distilled water to form a solution with the mass concentration of 2%, then adding a photoinitiator with the mass of 0.5% of that of the hydroxyethyl chitosan modified by the glycidyl methacrylate, magnetically stirring and uniformly mixing, and then irradiating by using ultraviolet light for 2 minutes to obtain hydroxyethyl chitosan hydrogel, namely chitosan derivative hydrogel; wherein the photoinitiator is 1-hydroxycyclohexyl phenyl ketone; the parameters of ultraviolet irradiation are as follows: the central wavelength is 365nm, and the light intensity is 1500mJ/cm2The irradiation distance was 15 cm.
Example 4
1) Dispersing chitosan with viscosity-average molecular weight of 500kDa and deacetylation degree of 93 percent in NaOH solution with mass concentration of 40 percent, wherein the mass ratio of the chitosan to the NaOH in the NaOH solution is 3: 5; adding an isopropanol solution of chloroethanol, and then heating to 65 ℃ for reflux reaction for 6 hours; wherein, the number of the chitosan repeating units is as follows: chloroethanol: the molar ratio of isopropanol was 1:6: 9.
2) After the reaction is finished, regulating the pH value to be neutral by hydrochloric acid, washing the obtained product for several times by using ethanol with the volume concentration of 80% and absolute ethanol, dissolving the product in water after vacuum drying, dialyzing for 48 hours by using a dialysis bag with the molecular weight cutoff of 3500Da, and then freeze-drying, specifically keeping the product at minus 20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the hydroxyethyl chitosan.
3) Preparing 0.2% hydroxyethyl chitosan solution, adding 0.7 times of glycidyl methacrylate of the number of repeating units of hydroxyethyl chitosan, adjusting the pH value to 8.0 by adopting 1mol/L NaOH solution, magnetically stirring for 4 hours at 75 ℃, and dialyzing for 3 days by using deionized water by adopting a dialysis bag with the molecular weight cutoff of 3500 Da; and finally, freeze-drying the dialysis product solution, specifically keeping the dialysis product solution at-20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the glycidyl methacrylate modified hydroxyethyl chitosan.
4) Dissolving hydroxyethyl chitosan modified by glycidyl methacrylate in distilled water to form a solution with the mass concentration of 3%, then adding a photoinitiator with the mass of 0.05% of that of the hydroxyethyl chitosan modified by the glycidyl methacrylate, magnetically stirring and uniformly mixing, and then irradiating by using ultraviolet light for 5 minutes to obtain hydroxyethyl chitosan hydrogel, namely chitosan derivative hydrogel; wherein the photoinitiator is 2-hydroxy-4' - (2-hydroxyethoxy) -2-methyl propiophenone; the parameters of ultraviolet irradiation are as follows: the central wavelength is 365nm, and the light intensity is 1300mJ/cm2The irradiation distance was 15 cm.
Example 5
1) Dispersing chitosan with the viscosity-average molecular mass of 700kDa and the deacetylation degree of more than 96% in a NaOH solution with the mass concentration of 50%, wherein the mass ratio of the chitosan to the NaOH in the NaOH solution is 3: 5; adding an isopropanol solution of chloroethanol, and then heating to 65 ℃ for reflux reaction for 5 hours; wherein, the number of the chitosan repeating units is as follows: chloroethanol: the molar ratio of isopropanol was 1:5: 8.
2) After the reaction is finished, regulating the pH value to be neutral by hydrochloric acid, washing the obtained product for several times by using ethanol with the volume concentration of 90% and absolute ethanol, dissolving the product in water after vacuum drying, dialyzing for 48 hours by using a dialysis bag with the molecular weight cutoff of 3500Da, and then freeze-drying, specifically keeping the product at minus 20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the hydroxyethyl chitosan.
3) Preparing 0.8% hydroxyethyl chitosan solution, adding 0.9 times of glycidyl methacrylate of the number of repeating units of hydroxyethyl chitosan, adjusting the pH value to 8.0 by adopting 1mol/L NaOH solution, magnetically stirring for 3 hours at 85 ℃, and dialyzing for 3 days by adopting a dialysis bag with the molecular weight cutoff of 3500Da through deionized water; and finally, freeze-drying the dialysis product solution, specifically keeping the dialysis product solution at-20 ℃ for 72 hours and 20 ℃ for 4 hours to obtain the glycidyl methacrylate modified hydroxyethyl chitosan.
4) Dissolving hydroxyethyl chitosan modified by glycidyl methacrylate in distilled water to form a solution with the mass concentration of 4%, then adding a photoinitiator with the mass of 0.4% of that of the hydroxyethyl chitosan modified by the glycidyl methacrylate, magnetically stirring and uniformly mixing, and irradiating by using ultraviolet light for 8 minutes to obtain hydroxyethyl chitosan hydrogel, namely chitosan derivative hydrogel; wherein the photoinitiator is 1-hydroxycyclohexyl phenyl ketone; the parameters of ultraviolet irradiation are as follows: the central wavelength is 365nm, and the light intensity is 1400mJ/cm2The irradiation distance was 15 cm.
Claims (2)
1. A preparation method of chitosan derivative hydrogel is characterized in that hydroxyethyl chitosan modified by glycidyl methacrylate is dissolved in distilled water to form a solution with the mass concentration of 1% -4%, then a photoinitiator with the mass concentration of 0.05% -0.5% of the hydroxyethyl chitosan modified by glycidyl methacrylate is added, and after uniform mixing, ultraviolet light is irradiated for 0.5-8 minutes to obtain hydroxyethyl chitosan hydrogel, namely the chitosan derivative hydrogel; the hydrogel has a communicated porous structure and the average pore diameter is 165 mu m;
wherein the photoinitiator is 2-hydroxy-4' - (2-hydroxyethoxy) -2-methyl propiophenone or 1-hydroxycyclohexyl phenyl ketone; the parameters of the ultraviolet irradiation are that the central wavelength is 365nm, and the light intensity is 1200mJ/cm2~ 1500 mJ/cm2The irradiation distance is 15 cm;
the preparation method of the glycidyl methacrylate modified hydroxyethyl chitosan comprises the following steps: preparing a hydroxyethyl chitosan solution with the mass concentration of 0.2-2%, then adding glycidyl methacrylate with the number of repeating units of the hydroxyethyl chitosan being 0.1-0.9 times, adjusting the pH value to 8.0 by using a NaOH solution with the concentration of 1mol/L, magnetically stirring for 3-8 hours at the temperature of 65-85 ℃, dialyzing for 3 days by using deionized water, wherein the cut-off molecular weight of a dialysis bag used for dialysis is 3500 Da; finally, freeze-drying the dialysis product solution, wherein the freeze-drying is carried out for 72 hours at the temperature of minus 20 ℃ and 4 hours at the temperature of 20 ℃ to obtain the glycidyl methacrylate modified hydroxyethyl chitosan;
the preparation method of the hydroxyethyl chitosan comprises the following steps:
1) dispersing chitosan into a NaOH solution with the mass concentration of 30-60%, wherein the mass ratio of the chitosan to the NaOH is 3: 5; adding an isopropanol solution of chloroethanol, and then heating to 65 ℃ for reflux reaction for 4-7 hours; wherein, the number of the chitosan repeating units is as follows: chloroethanol: the molar ratio of the isopropanol is 1:3: 5-1: 6: 9;
2) after the reaction is finished, regulating the pH value to be neutral by hydrochloric acid, washing the obtained product for several times by using 75-90% volume concentration ethanol and absolute ethanol, dissolving the product in water after vacuum drying, dialyzing for 48 hours, and freeze-drying to obtain hydroxyethyl chitosan;
the viscosity-average molecular weight of the chitosan is 300-1000 kDa, and the deacetylation degree is more than 90 percent.
2. A chitosan derivative hydrogel produced according to the method of claim 1, wherein the hydrogel has a interconnected porous structure and an average pore size of 165 μm.
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