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Silk fibroin base integrated osteochondral two-layer bracket, preparation and application thereof

A technology of silk fibroin and protein scaffolds, applied in coatings, prostheses, medical science, etc., can solve the problems of application limitations, poor osteoinductivity and osteoconductivity, etc., achieve good biocompatibility, and facilitate cell adhesion , good bone repair effect

Active Publication Date: 2013-04-10
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its disadvantage is that it is difficult to form weakly crystalline hydroxyapatite. Although sufficient calcium sources can be obtained, the osteoinductivity and osteoconductivity are poor, so the application is greatly limited.

Method used

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  • Silk fibroin base integrated osteochondral two-layer bracket, preparation and application thereof
  • Silk fibroin base integrated osteochondral two-layer bracket, preparation and application thereof
  • Silk fibroin base integrated osteochondral two-layer bracket, preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Preparation of silk fibroin solution: a) Degumming: put 100g of silkworm silk (Zhejiang Huazhi Silk Co., Ltd., 5A grade) into 5L of 2M sodium carbonate aqueous solution, bathe in 96°C water for 30min, wash with purified water, and repeat the process 3 times, remove the sericin, leave the silk fibroin, and dry the silk fibroin at 50°C to obtain 70g of dried silk fibroin for later use; b) Dissolving: the above-mentioned dried silk fibroin is Dissolve in 9.3M lithium bromide (LiBr) aqueous solution at a volume ratio of 0.2:1, bathe in water at 60°C for 90 minutes until the silk fibroin is fully dissolved, and obtain a mixed solution containing silk fibroin and a small amount of insoluble particles; c) Dialysis: Dilute the mixed solution with Dialyze the regenerated cellulose dialysis bag (molecular weight cut off 4000 Daltons), dialyze 12 times with sterile purified water of 10 times the volume of the mixed solution in 3 days, remove LiBr ions in the solution, and obtain th...

Embodiment 2

[0042] Example 2: Silk fibroin-based integrated osteochondral double-layer scaffold

[0043] (1) Silk fibroin solution: take 100 mg of silk fibroin after freeze-drying by the method of Example 1, dissolve it in deionized water at 4° C. for 3 days, and prepare 2 ml of a 50 mg / ml silk fibroin solution.

[0044] (2) Mold making: Use 5 1ml disposable volumetric syringes (both outer diameters are 6.5mm), including the barrel of the syringe and the push rod with the piston, as the mold; and the barrel needle of the disposable volumetric syringe The end of the seat is sintered and closed, and the mold cylinder is made of through holes dug on the closed surface. The hole diameter is equivalent to that of sodium chloride particles (ie, 45-50 μm), and the hole distance is 2 mm.

[0045] (3) Material compression: Pour 0.28g (0.2ml) of pre-ground NaCl particles with a particle size of 45-50μm into the mold, and then add 0.2ml of the silk fibroin solution in step (1). Quickly push in the ...

Embodiment 3

[0049] Example 3: Silk fibroin-based integrated osteochondral double-layer scaffold

[0050] (1) Silk fibroin solution: the silk fibroin prepared in Example 1 was dissolved in deionized water at 4° C. for 3 days to prepare 15 ml of a 60 mg / ml silk fibroin solution.

[0051] (2) Mold making: Use four 10ml disposable volumetric syringes (outside diameter of 16.5mm), including the cylinder of the syringe and the push rod with the piston, as the mold; and the needle seat of the cylinder of the disposable volumetric syringe This end is sintered closed and the said mold cylinder is made by digging through holes on the closed surface, the hole diameter is equivalent to that of sodium chloride particles (150-200 μm), and the hole distance is 3mm.

[0052] (3) Material compression: Pour 2.8g (2ml) of pre-ground NaCl particles with a particle size of 150-200μm into each mold, and then add 3ml of the silk fibroin solution prepared in step (1) at a rate of 3cm / Push in the solution at a ...

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Abstract

The invention discloses a silk fibroin base integrated osteochondral two-layer bracket, preparation and application thereof. The preparation of the silk fibroin base integrated osteochondral two-layer bracket is carried out by the following steps of: respectively soaking parts of 1 / 2-3 / 4 height of a three-dimensional silk fibroin bracket in a calcium chloride ethanol solution, absolute ethyl alcohol, a dipotassium phosphate aqueous solution and de-ionized water; repeating the above operation for 1-15 times, taking the silk fibroin bracket finally soaked by the de-ionized water to soak in the calcium chloride ethanol solution for 5-30 min and subsequently using the de-ionized water to soak the silk fibroin bracket for 3-10 min and pre-calcify the three-dimensional silk fibroin bracket; then soaking the pre-calcified part of the three-dimensional silk fibroin bracket in a bionic calcium ion buffer solution and culturing for 4-8 days at a constant temperature of 37 DEG C to obtain the silk fibroin base integrated osteochondral two-layer bracket. In the pre-calcification process of the invention, the silk fibroin bracket can obtain a nucleating locus of hydroxyapatite crystals; and evenly dispersed weak crystallization nano hydroxyapatites are formed on the surfaces of the porous channels of the bracket by culturing the bracket in the bionic calcium ion buffer solution.

Description

(1) Technical field [0001] The present invention relates to a preparation method of a double-layer scaffold material that can be used for osteochondral repair, in particular to a silk fibroin-based integrated osteochondral double-layer scaffold and its preparation method and application, that is, a bionic construction with silk fibroin as a matrix material A Structurally Integrated Approach to Bilayer Scaffold Materials for Osteochondral Repair. (2) Background technology [0002] Osteochondral defects caused by traffic accidents, sports trauma, and various arthritis diseases have brought great pain and heavy economic burden to patients and their families, not only affecting the quality of life of patients, but also increasing social pressure. At present, autologous osteochondral transplantation and allogeneic or heterogeneous osteochondral transplantation are generally used in the clinical treatment of articular osteochondral defects. The former has limited sources and crea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/40A61L27/22A61L27/32A61L31/12
Inventor 徐国伟赵洪石陈隆坤欧阳宏伟
Owner ZHEJIANG UNIV
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