CN113198049B - Myocardial repair hydrogel and preparation method thereof - Google Patents
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Abstract
The invention relates to the technical field of bionic materials, in particular to a cardiac muscle repair hydrogel and a preparation method thereof. The myocardial repair hydrogel comprises the following components in parts by weight: 10-70 parts of glycidyl methacrylate modified glucan, 3-10 parts of polypyridine and 10 parts of vascular endothelial factor‑6～10‑5And (4) portions are obtained. The myocardial repair hydrogel has injectability, has the conductive characteristic similar to that of myocardial cells, has better compatibility, can attract the aggregation of vascular endothelial cells and platelets, and has obvious promotion effect on the proliferation of the myocardial cells.
Description
Technical Field
The invention relates to the technical field of bionic materials, in particular to a myocardial repair hydrogel and a preparation method thereof.
Background
The native myocardium is an electrically active tissue in which purkinje fibers with electrical conduction function are distributed. Electrical signals from the sinoatrial node, when they pass through the myocardium, induce cardiomyocyte synchronous contraction through "excitatory-contractile" coupling. Meanwhile, the myocardial tissue has excellent mechanical strength and can support continuous contraction and relaxation during the heart beating. Therefore, the scaffold material for engineering myocardial tissue construction should have both excellent electrical conductivity and elastic modulus.
The biological scaffold plays a critical role in tissue engineering reconstruction, can be used as a drug carrier, creates a bionic biological microenvironment for cells, is beneficial to the transmission of nutrient substances and the transportation of cell wastes, has higher requirements on the structure of the scaffold for partial deletion or damage, and has higher requirements on the convenience of operation. The requirements of the scaffold for repairing myocardial damage not only need to be favorable for adhesion and regeneration of myocardial cells, but also need to ensure that the damaged part does not expand due to stress action, so that the requirement on the contact degree of the scaffold material and the shape of the damaged part is higher, and more complete and synchronous shape recovery is achieved.
The hydrogel is a polymer which has hydrophilic groups, can swell in water and is insoluble in water and has a cross-linked three-dimensional network structure, is an important functional polymer material, and is one of the research hotspots of the current material science. The hydrogel contains hydrophilic groups, can absorb a large amount of water to swell in water, mostly contains higher water content and lower modulus, and can keep the solid property. The hydrogel material can absorb and retain a large amount of water, is beneficial to the transportation of cell nutrients and metabolites, has adjustable physical and chemical properties, and is widely researched in myocardial tissue engineering, but the hydrogel with a single component is usually an electrical insulating material and is not beneficial to the transmission of intercellular electrical signals.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide the myocardial repair hydrogel and the preparation method thereof.
In order to achieve the purpose, the invention adopts the technical scheme that:
the invention provides a myocardial repair hydrogel which comprises the following components in parts by weight:
10-70 parts of glycidyl methacrylate modified glucan, 3-10 parts of polypyridine and 10 parts of vascular endothelial factor-6～10-5And (4) portions are obtained.
The glucan modified by glycidyl methacrylate esterification has a double bond part of glycidyl methacrylate molecules, can generate self-crosslinking reaction under the action of a photoinitiator, does not need an additional crosslinking agent, and reduces the toxic effect; the polypyridine has good biocompatibility and can provide a conductive function, the function of the dextran modified by glycidyl methacrylate esterification is not limited after the polypyridine is used together with the dextran modified by glycidyl methacrylate esterification, a coordination effect can be achieved, the vascular endothelial factor can provide the cell compatibility of the hydrogel under a mild environment, the aggregation of vascular endothelial cells and platelets can be caused, and the further recovery of the myocardial function is facilitated. Through a large amount of researches and experiments, the inventor of the invention finds that the prepared hydrogel has an obvious conductive function by compounding the glycidol methacrylate modified glucan, the polypyridine and the vascular endothelial factor, can provide a signal transmission medium among myocardial cells, has better cell compatibility, and has an obvious promotion effect on the proliferation of the myocardial cells.
As a preferred embodiment of the myocardial repair hydrogel, the myocardial repair hydrogel comprises the following components in parts by weight:
30-60 parts of glycidyl methacrylate esterification modified glucan, 3-7 parts of polypyridine and 10 parts of vascular endothelial factor-6～10-5And (4) portions.
As a preferred embodiment of the myocardial repair hydrogel, the myocardial repair hydrogel comprises the following components in parts by weight:
50 parts of dextran modified by glycidyl methacrylate esterification, 3 parts of polypyridine and 10 parts of vascular endothelial factor-5And (4) portions are obtained.
In the technical scheme of the invention, the specific components in parts by weight are matched with each other, so that the conductive function is optimal, the aggregation of vascular endothelial cells and platelets is better attracted, and the proliferation of myocardial cells is better and obviously promoted.
In a preferred embodiment of the myocardial repair hydrogel of the present invention, the glycidyl methacrylate-modified dextran is obtained by reacting dextran with glycidyl methacrylate.
In a preferred embodiment of the myocardial repair hydrogel according to the present invention, the dextran is an a linear dextran.
In the technical scheme of the invention, the alpha linear glucan has good shear thinning characteristic and stability, the structure of the alpha linear glucan is similar to that of mucopolysaccharide in a cell matrix, and the alpha linear glucan has no cytotoxicity and better safety.
As a preferred embodiment of the myocardial repair hydrogel, the preparation method of the glycidyl methacrylate esterification modified glucan comprises the following steps:
dissolving glucan in dimethyl sulfoxide to obtain a solution with the mass concentration of 1-5%; then adding 4-methyl pyrrolidone and glycidyl methacrylate into the solution, uniformly mixing, and reacting at room temperature for 24-48 hours; after the reaction is finished, adding sodium hydroxide, filtering, precipitating, washing, and freeze-drying to obtain the product.
The second purpose of the invention is to provide a preparation method of the myocardial repair hydrogel, which comprises the following steps:
s1, dissolving dextran modified by glycidyl methacrylate esterification in water, adding polypyridine and vascular endothelial factors, and uniformly mixing to obtain a mixed solution A;
s2, dissolving a photoinitiator in N-methyl pyrrolidone, and adding the photoinitiator into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by using ultraviolet light to obtain the myocardial repair hydrogel.
The invention adopts controllable photocuring time, so that the hydrogel has better operation convenience when repairing myocardial damage, and can improve the filling degree of the damaged part.
In a preferred embodiment of the method for preparing the myocardial repair hydrogel, the photoinitiator is photoinitiator I2959.
In the technical scheme of the invention, under the action of the photoinitiator I2959, the dextran modified by glycidyl methacrylate esterification can generate self-crosslinking reaction, no additional crosslinking agent is needed, the toxic effect is reduced, and the safety is high.
As a preferred embodiment of the method for preparing the myocardial repair hydrogel, the wavelength of the ultraviolet light is 365 nm.
As a preferred embodiment of the preparation method of the myocardial repair hydrogel, the ultraviolet irradiation time is 1-3 min.
Compared with the prior art, the invention has the following beneficial effects:
the myocardial repair hydrogel provided by the invention has an obvious conductive function, and is beneficial to the transmission of electric signals between myocardial cells; the hydrogel has better cell compatibility, can attract the aggregation of vascular endothelial cells and platelets, and has obvious promotion effect on the proliferation of myocardial cells.
Drawings
FIG. 1 is a graph showing the effect of using the myocardial repair hydrogel of the present invention on myocardial cell activity.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to the accompanying drawings and specific embodiments.
In the following examples and comparative examples, the starting materials used were all commercially available, unless otherwise specified.
Preparation of dextran modified by glycidyl methacrylate esterification:
the preparation method of the dextran modified by glycidyl methacrylate esterification comprises the following steps:
dissolving 5 parts by weight of dextran in 200 ml of anhydrous dimethyl sulfoxide solvent to obtain a solution; adding 1 part by weight of 4-methyl pyrrolidone and 2 parts by weight of glycidyl methacrylate into the solution, uniformly mixing, and reacting at room temperature for 24-48 hours; and after the reaction is finished, dropwise adding sodium hydroxide to generate a precipitate, filtering the precipitate, washing the precipitate with deionized water, washing the precipitate with hydrochloric acid to be neutral, and freeze-drying the neutral solution to obtain the glycidol methacrylate esterification modified glucan.
Example 1 myocardial repair hydrogel of the invention
The myocardial repair hydrogel comprises the following components in parts by weight:
50 parts of dextran modified by glycidyl methacrylate esterification, 3 parts of polypyridine and 10 parts of vascular endothelial factor-5And (4) portions are obtained.
The preparation method of the myocardial repair hydrogel comprises the following steps:
s1, dissolving glycidol methacrylate modified glucan in 200 ml of water, adding polypyridine and vascular endothelial factor, and uniformly mixing to obtain a mixed solution A;
s2, dissolving a photoinitiator I2959 in N-methylpyrrolidone to obtain a solution with the mass concentration of 0.1%, and adding 20 microliters of the solution into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by adopting ultraviolet light with the wavelength of 365nm for 3min to obtain the myocardial repair hydrogel.
Example 2 myocardial repair hydrogel of the invention
The myocardial repair hydrogel comprises the following components in parts by weight:
30 parts of dextran modified by glycidyl methacrylate esterification, 3 parts of polypyridine and 10 parts of vascular endothelial factor-5And (4) portions are obtained.
The preparation method of the myocardial repair hydrogel comprises the following steps:
s1, dissolving glycidol methacrylate modified glucan in 200 ml of water, adding polypyridine and vascular endothelial factor, and uniformly mixing to obtain a mixed solution A;
s2, dissolving the photoinitiator I2959 in N-methyl pyrrolidone to obtain a solution with the mass concentration of 0.1%, and adding 20 microliters of the solution into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by adopting ultraviolet light with the wavelength of 365nm for 3min to obtain the myocardial repair hydrogel.
Example 3 myocardial repair hydrogel of the invention
The myocardial repair hydrogel comprises the following components in parts by weight:
10 parts of dextran modified by glycidyl methacrylate esterification, 3 parts of polypyridine and 10 parts of vascular endothelial factor-5And (4) portions are obtained.
The preparation method of the myocardial repair hydrogel comprises the following steps:
s1, dissolving glycidol methacrylate modified glucan in 200 ml of water, adding polypyridine and vascular endothelial factor, and uniformly mixing to obtain a mixed solution A;
s2, dissolving the photoinitiator I2959 in N-methyl pyrrolidone to obtain a solution with the mass concentration of 0.1%, and adding 20 microliters of the solution into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by adopting ultraviolet light with the wavelength of 365nm for 3min to obtain the myocardial repair hydrogel.
Example 4 myocardial repair hydrogel of the invention
The myocardial repair hydrogel comprises the following components in parts by weight:
50 parts of dextran modified by glycidyl methacrylate esterification, 3 parts of polypyridine and 10 parts of vascular endothelial factor-5And (4) portions are obtained.
The preparation method of the myocardial repair hydrogel comprises the following steps:
s1, dissolving glycidol methacrylate modified glucan in 200 ml of water, adding polypyridine and vascular endothelial factor, and uniformly mixing to obtain a mixed solution A;
s2, dissolving the photoinitiator I2959 in N-methyl pyrrolidone to obtain a solution with the mass concentration of 0.1%, and adding 20 microliters of the solution into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by adopting ultraviolet light with the wavelength of 365nm for 1min to obtain the myocardial repair hydrogel. Only different from example 1 is that step S3 was irradiated for 1 min.
Example 5 myocardial repair hydrogel of the invention
The myocardial repair hydrogel comprises the following components in parts by weight:
60 parts of dextran modified by glycidyl methacrylate esterification, 7 parts of polypyridine and 10 parts of vascular endothelial factor-6And (4) portions.
The preparation method of the myocardial repair hydrogel comprises the following steps:
s1, dissolving glycidyl methacrylate esterification modified glucan in 200 ml of water, adding polypyridine and vascular endothelial factor, and uniformly mixing to obtain a mixed solution A;
s2, dissolving the photoinitiator I2959 in N-methyl pyrrolidone to obtain a solution with the mass concentration of 0.1%, and adding 20 microliters of the solution into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by adopting ultraviolet light with the wavelength of 365nm for 3min to obtain the myocardial repair hydrogel.
Example 6 myocardial repair hydrogel of the invention
The myocardial repair hydrogel comprises the following components in parts by weight:
70 parts of dextran modified by glycidyl methacrylate esterification, 10 parts of polypyridine and 10 parts of vascular endothelial factor-5And (4) portions.
The preparation method of the myocardial repair hydrogel comprises the following steps:
s1, dissolving glycidol methacrylate modified glucan in 200 ml of water, adding polypyridine and vascular endothelial factor, and uniformly mixing to obtain a mixed solution A;
s2, dissolving the photoinitiator I2959 in N-methyl pyrrolidone to obtain a solution with the mass concentration of 0.1%, and adding 20 microliters of the solution into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by adopting ultraviolet light with the wavelength of 365nm for 3min to obtain the myocardial repair hydrogel.
Comparative example 1
Similar to example 1, except that the myocardial repair hydrogel did not contain vascular endothelial factor, the remaining raw materials were the same as in example 1.
Comparative example 2
Similar to example 1, except that the myocardium repair hydrogel does not contain polypyridine, the rest of the raw materials are the same as example 1.
Test example, myocardial repair hydrogel Performance test
The in vitro physicochemical properties including mechanical strength and conductivity of the myocardial repair hydrogels obtained in the above examples 1 to 6 and comparative examples 1 to 2 were evaluated under the same environmental conditions by using the GB/T16886.5-2017 standard.
The proliferation of cardiomyocytes by the myocardial repair hydrogel of the present invention was measured according to the following procedure.
1) A suspension of H9C2 cells was prepared.
2) Cell suspensions were seeded in 96-well plates, 5000 cells per well, with at least three replicates per group.
3) The plates were pre-incubated in an incubator for 12,24 and 36 hours (37 ℃, 5% CO)2)。
4) Each well was filled with 10. mu.L of CCK-8 solution.
5) The culture plate is put into an incubator to be incubated for 1-4 h.
6) The absorbance (OD) at 450nm was measured with a microplate reader.
The specific results of the experiment are shown in tables 1-2 and FIG. 1:
TABLE 1
TABLE 2
| Group of | Number of parallel groups | Electrical conductivity (10)-4S/cm) |
| Example 1 | 3 | 27.3±2 |
| Example 2 | 3 | 24.8±0.8 |
| Example 3 | 3 | 20.4±0.6 |
| Example 4 | 3 | 25.3±0.6 |
| Example 5 | 3 | 58.2±0.4 |
| Example 6 | 3 | 95.2±0.7 |
| Comparative example 1 | 3 | 26.3±0.3 |
| Comparative example 2 | 3 | 3.4±0.3 |
As can be seen from the data in tables 1 and 2, the hydrogel for myocardial repair prepared according to the present invention has better mechanical strength and electrical conductivity. Compared with the prior art, the hydrogel does not contain polypyridine, the conductivity of the hydrogel is the lowest, and the inventor of the invention finds that the prepared hydrogel has an obvious conductive function by compounding the glycidol methacrylate modified dextran, the polypyridine and the vascular endothelial factor through a large amount of research and experiments, can provide a signal transmission medium among myocardial cells, and has better compatibility of the cells.
Referring to fig. 1, the myocardial repair hydrogel prepared by the present invention has an obvious effect of promoting proliferation of myocardial cells, and from the comprehensive performance comparison, embodiment 1 is the best embodiment, which is beneficial to the transmission of electrical signals between myocardial cells; the hydrogel has good cell compatibility, can attract the aggregation of vascular endothelial cells and platelets, and has obvious promotion effect on the proliferation of myocardial cells.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (7)
1. The myocardial repair hydrogel is characterized by comprising the following components in parts by weight:
10-70 parts of glycidyl methacrylate modified glucan, 3-10 parts of polypyridine and 10 parts of vascular endothelial factor-6～10-5Preparing;
the preparation method of the glycidyl methacrylate esterification modified glucan comprises the following steps:
dissolving glucan in dimethyl sulfoxide to obtain a solution with the mass concentration of 1-5%; then adding 4-methyl pyrrolidone and glycidyl methacrylate into the solution, uniformly mixing, and reacting at room temperature for 24-48 hours; after the reaction is finished, adding sodium hydroxide, filtering, precipitating, washing, and freeze-drying to obtain the product;
the preparation method of the myocardial repair hydrogel comprises the following steps:
s1, dissolving dextran modified by glycidyl methacrylate esterification in water, adding polypyridine and vascular endothelial factors, and uniformly mixing to obtain a mixed solution A;
s2, dissolving a photoinitiator in N-methyl pyrrolidone, and adding the photoinitiator into the mixed solution A to obtain a mixed solution B;
and S3, irradiating the mixed solution B by using ultraviolet light to obtain the myocardial repair hydrogel.
2. The myocardial repair hydrogel of claim 1, comprising the following components in parts by weight:
30-60 parts of glycidyl methacrylate esterification modified glucan, 3-7 parts of polypyridine and 10 parts of vascular endothelial factor-6～10-5And (4) portions are obtained.
3. The myocardial repair hydrogel of claim 2, comprising the following components in parts by weight:
50 parts of dextran modified by glycidyl methacrylate esterification, 3 parts of polypyridine and 10 parts of vascular endothelial factor-5And (4) portions are obtained.
4. The myocardial repair hydrogel of claim 1, wherein the dextran is an a linear dextran.
5. The myocardial repair hydrogel of claim 1, wherein the photoinitiator is photoinitiator I2959.
6. The myocardial repair hydrogel of claim 5, wherein the ultraviolet light has a wavelength of 365 nm.
7. The myocardial repair hydrogel according to claim 5, wherein the UV irradiation time is 1 to 3 min.
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