Hydrogel loaded with bone morphogenetic protein, preparation method and application
A hydrogel and protein technology, applied in the fields of pharmaceutical formula, medical science, prosthesis, etc., can solve the problems of poor biocompatibility, immunogenicity, insufficient reaction conditions, cytotoxicity, etc., shorten the molding time and improve the utilization rate, promote the effect of repair
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[0035] The invention provides a method for preparing a hydrogel loaded with bone morphogenic protein, comprising: mixing and reacting a cross-linking agent, a water-soluble natural degradable polymer, and bone morphogenic protein in a liquid for injection or an aqueous buffer solution to obtain a bone-loaded gel. A protein hydrogel is formed.
[0036] The bone morphogenic protein includes more than 30 related proteins, such as BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, etc., the bone morphogenic protein described in this application It is one or a mixture of two or more of the 30 related proteins.
[0037] The crosslinking agent is 1,4 butanediol diglycidyl ether, glutaraldehyde, N-hydroxysuccinimide ester, carbodiimide, genipin, cystamine, hexamethylene isodicyanate, azide One or more of nitrogen diphenylphosphine, polyethylene glycol diacrylate, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
[0038] The water-soluble natural degradable polymer is one o...
Embodiment 1
[0068] (1) Sterilization of raw materials
[0069] The polyglutamate powder with a weight-average molecular weight of 1.5 million Daltons was sterilized by irradiation with gamma-rays at a sterilization dose of 6 kGy.
[0070] (2) Preparation of hydrogel
[0071] The following procedures were performed under sterile conditions.
[0072] Dissolve 0.01 mL of 1,4-butanediol diglycidyl ether in 5 mL of double-distilled water, and add 2.5 mg of BMP-2 protein. Weigh 0.25 g of the sterilized polyglutamate powder with a weight-average molecular weight of 1.5 million Daltons, add it to the above solution, stir evenly, let stand for a while until the bubbles disappear, and keep warm at 37°C for 10 hours.
[0073] The release rate of bone morphogenetic protein in the products prepared by this method was 3.17%, 14.84%, 32.06%, 50.15%, 65.02%, 71.91%, 78.73% respectively on the 2d, 5d, 10d, 15d, 20d, 25d, and 30d; The porosity (area / total area occupied by pores) of the product reaches 9...
Embodiment 2
[0075] (1) Sterilization of raw materials
[0076] Hyaluronic acid with a weight-average molecular weight of 2.5 million Daltons was sterilized by irradiation with gamma-rays, and the sterilization dose was 10 kGy;
[0077] (2) Preparation of hydrogel
[0078] The following procedures were performed under sterile conditions.
[0079] Dissolve 0.05 mL of glutaraldehyde in 5 mL of borate buffer, and add 25 mg of BMP-2. Weigh 1.0 g of the sterilized hyaluronic acid with a weight-average molecular weight of 2.5 million Daltons, add it to the above solution, stir evenly, let it stand still for a while until the bubbles disappear, and keep warm at 60° C. for 2 hours.
[0080] The release rate of the bone morphogenic protein of the product prepared by this method on the 2d, 5d, 10d, 15d, 20d, 25d, and 30d was 2.00%, 13.00%, 30.00%, 48.00%, 63.0%, 70.0%, and 76.0% respectively; The porosity (area / total area occupied by pores) of the product reaches 97%; the cell proliferation rate ...
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