阅读说明：本技术 具有自修复性能的仿生纤维网水凝胶的制备方法及其应用 (Preparation method and application of bionic fiber mesh hydrogel with self-repairing performance ) 是由 李凯 曾艳 梁峰 于 2020-06-28 设计创作，主要内容包括：本发明提供了一种具有自修复性能的仿生纤维网水凝胶的制备方法及其应用,制备方法包括如下步骤：1)将海藻酸钠氧化为氧化海藻酸钠；2)以0.137 mol/L的己二酸作为溶剂配制含量为0.01%的壳聚糖溶液,将壳聚糖溶液缓慢滴加入聚丙烯酸溶液中,滴加完成后室温下反应1 h,待反应结束后先将溶液在-80℃冷冻,然后解冻离心洗涤,最后冷冻干燥得到壳聚糖纤维；3)以生理盐水为溶剂配制氧化海藻酸钠溶液和含有壳聚糖纤维的羧甲基壳聚糖溶液,氧化海藻酸钠溶液和羧甲基壳聚糖溶液按体积比为1:2混合进行反应得到多级次纤维网水凝胶。纤维网水凝胶具有可修复性能,与天然细胞外基质结构相似,在三维细胞培养、软组织修复等组织工程领域具有很好的应用前景。(The invention provides a preparation method and application of bionic fiber mesh hydrogel with self-repairing performance, wherein the preparation method comprises the following steps: 1) oxidizing sodium alginate into oxidized sodium alginate; 2) preparing a chitosan solution with the content of 0.01% by taking 0.137 mol/L adipic acid as a solvent, slowly dropwise adding the chitosan solution into a polyacrylic acid solution, reacting at room temperature for 1 h after dropwise adding is completed, freezing the solution at-80 ℃ after the reaction is completed, then thawing, centrifuging, washing, and finally freeze-drying to obtain chitosan fibers; 3) preparing an oxidized sodium alginate solution and a carboxymethyl chitosan solution containing chitosan fibers by using normal saline as a solvent, and mixing the oxidized sodium alginate solution and the carboxymethyl chitosan solution according to the volume ratio of 1:2 for reaction to obtain the multi-level fiber mesh hydrogel. The fiber mesh hydrogel has repairable performance, is similar to a natural extracellular matrix structure, and has good application prospect in the tissue engineering fields of three-dimensional cell culture, soft tissue repair and the like.)

1. A preparation method of bionic fiber mesh hydrogel with self-repairing performance is characterized by comprising the following steps:

1) oxidizing sodium alginate into oxidized sodium alginate by using sodium periodate as an oxidizing agent;

2) preparing 0.01 percent chitosan solution by taking 0.137 mol/L adipic acid as a solvent, preparing 1 percent polyacrylic acid solution with volume fraction by taking distilled water as a solvent, slowly dripping the chitosan solution into the polyacrylic acid solution, reacting for 1 h at room temperature after dripping is finished, freezing the solution at-80 ℃ after the reaction is finished, then unfreezing, centrifuging, washing, and finally freeze-drying to obtain chitosan fibers;

3) preparing an oxidized sodium alginate solution and a carboxymethyl chitosan solution containing chitosan fibers by using normal saline as a solvent, wherein the concentration of the oxidized sodium alginate solution is 100 mg/mL, the concentration of the carboxymethyl chitosan solution is 25 mg/mL, and the oxidized sodium alginate solution and the carboxymethyl chitosan solution are mixed according to the volume ratio of 1:2 for reaction to obtain the multi-level fiber mesh hydrogel.

2. The method for preparing a biomimetic fiber mesh hydrogel with self-repairing properties according to claim 1, wherein the oxidation degree of oxidized sodium alginate is 60%.

3. The method for preparing a biomimetic fiber mesh hydrogel with self-repairing properties according to claim 1, wherein the final volume ratio of the chitosan solution to the polyacrylic acid solution in the process of preparing the chitosan fiber is 1: 3.

4. The method for preparing a biomimetic fiber mesh hydrogel with self-repairing properties according to claim 1, wherein the chitosan fiber concentration in the carboxymethyl chitosan solution is 1 mg/mL.

5. The application of the bionic fiber mesh hydrogel with self-repairing performance is characterized in that the bionic fiber mesh hydrogel is applied to cell culture or soft tissue repair.

Technical Field

The invention belongs to the field of biological materials, and relates to a preparation method and application of a bionic fiber mesh hydrogel with self-repairing performance.

Background

Extracellular matrix (ECM) is widely present in various extracellular spaces, even penetrating into small spaces of muscles, tendons, and other tissues, as a representative of biological soft matter. The layered porous medium consisting of the fiber mesh consisting of fiber bundles (micron scale) and gel-like matrix (nanometer scale) not only enables the ECM to fix organs on epithelial tissues and attach to other underlying tissues, but also enables more efficient long-range transmission of molecular signals. This multilevel structure of ECM directly affects its physical properties and response to mechanical stress, its fiber content, structure and arrangement are different in different organ tissues and different pathophysiological states, and influences life processes through changes in the multilevel state. For example, the fibrous structure of the ECM affects the transmission of growth factor signals. Growth factors can bind to the surface of ECM fibers, bringing them into close proximity to the cell surface. In this way the ECM promotes the interaction between growth factors and the cell surface.

Conventional hydrogels have many limitations in ECM simulation, firstly their structure is too simple compared to the complex structure of native tissue, and secondly they lack mechanical stimulation to induce changes in cell phenotype and thus direct the formation of corresponding tissue structures.

Disclosure of Invention

The invention aims to provide a preparation method and application of bionic fiber mesh hydrogel with self-repairing performance.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the preparation method of the bionic fiber mesh hydrogel with the self-repairing performance comprises the following steps:

1) oxidizing sodium alginate into oxidized sodium alginate by using sodium periodate as an oxidizing agent;

2) preparing 0.01 percent chitosan solution by taking 0.137 mol/L adipic acid as a solvent, preparing 1 percent polyacrylic acid solution with volume fraction by taking distilled water as a solvent, slowly dripping the chitosan solution into the polyacrylic acid solution, reacting for 1 h at room temperature after dripping is finished, freezing the solution at-80 ℃ after the reaction is finished, then unfreezing, centrifuging, washing, and finally freeze-drying to obtain chitosan fibers;

3) preparing an oxidized sodium alginate solution and a carboxymethyl chitosan solution containing chitosan fibers by using normal saline as a solvent, wherein the concentration of the oxidized sodium alginate solution is 100 mg/mL, the concentration of the carboxymethyl chitosan solution is 25 mg/mL, and the oxidized sodium alginate solution and the carboxymethyl chitosan solution are mixed according to the volume ratio of 1:2 for reaction to obtain the multi-level fiber mesh hydrogel.

In the method, the oxidation degree of the oxidized sodium alginate is 60%.

In the method, the final volume ratio of the chitosan solution to the polyacrylic acid solution in the chitosan fiber preparation process is 1: 3.

In the method, the concentration of chitosan fibers in the carboxymethyl chitosan solution is 1 mg/mL.

The invention also provides a bionic fiber net hydrogel with self-repairing performance, which can be applied to cell culture or soft tissue repair.

The chitosan fiber (CMF) of the present invention is a rod-like fiber having a diameter of about 1 to several tens of micrometers.

The implementation of the invention has the following beneficial effects:

1. according to the invention, Oxidized Sodium Alginate (OSA) and carboxymethyl chitosan (CMCS) containing chitosan fiber (CMF) are mixed and crosslinked according to the Schiff base reaction principle to prepare the fiber hydrogel (OSA-CMCS-CMF) with high similarity to the ECM structure. The fiber mesh hydrogel has repairable performance, is similar to a natural extracellular matrix structure, and has good application prospect in the tissue engineering fields of three-dimensional cell culture, soft tissue repair and the like.

2. The hydrogel obtained by crosslinking the Schiff base dynamic covalent bonds has a dynamic self-healing effect, and has certain pH response capability due to the acid sensitivity of Schiff base reaction. After the CMF is added, the microstructure of the hydrogel is greatly changed, so that the hydrogel has a reticular fiber structure, and the reticular fiber has high similarity with the rat connective hoof tissue. The hydrogel is used as a substrate for two-dimensional cell culture, excellent biocompatibility is shown, and cells can migrate from the surface of the hydrogel to the interior of the hydrogel and aggregate on the surface of fibers. The results show that the hydrogel disclosed by the invention is not only similar to the ECM in structure, but also good in cell compatibility, and has a good application prospect in the field of tissue engineering.

3. The invention provides a brand-new strategy of imitating the ECM hydrogel, which is simpler and easier to implement, namely, the fiber hydrogel is prepared by mixing the gel after the fiber is prepared by electrostatic self-assembly.

4. The selected sodium alginate and chitosan are natural macromolecular polysaccharides and have good biocompatibility, so that the hydrogel prepared by the invention has good biocompatibility.

5. The material selected by the invention is simple and easy to obtain, has low price and is easy to realize industrial application.

Drawings

FIG. 1 is a flow chart of the preparation of a biomimetic fibrous web hydrogel of example 1 of the present invention;

FIG. 2 is a scanning electron micrograph and a transmission electron micrograph of a chitosan fiber (CMF) of example 1 of the present invention;

FIG. 3 is a graph showing the self-healing effect of the hydrogel of example 1 of the present invention;

FIG. 4 is a scanning electron micrograph of rat colonic tissue (left) and a hydrogel of example 1 of the present invention (right);

FIG. 5 shows an internal scanning electron micrograph (left) of a hydrogel of example 1 of the present invention after cell culture and after lateral cutting, and a bright field micrograph (upper right) and a fluorescence micrograph (lower right) under an optical microscope.

Detailed Description

The technical solution of the present invention is described below with reference to the accompanying drawings and examples.