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Preparation method of intra-fibrous biomimetic mineralized collagen membrane capable of promoting osteogenic differentiation of cells

A biomimetic mineralization and osteogenic differentiation technology, applied in tissue regeneration, medical science, prostheses, etc., can solve problems such as bionics, achieve good cell adhesion, good biocompatibility, and promote cell osteogenic differentiation

Active Publication Date: 2020-03-24
THE FIRST AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Extrafibrous mineralization can biomime the chemical composition and a small part of the structure of the extracellular matrix of bone tissue, but the materials prepared by this method cannot achieve real biomimesis of the microstructure, formation process and microenvironment of the bone tissue matrix

Method used

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  • Preparation method of intra-fibrous biomimetic mineralized collagen membrane capable of promoting osteogenic differentiation of cells
  • Preparation method of intra-fibrous biomimetic mineralized collagen membrane capable of promoting osteogenic differentiation of cells
  • Preparation method of intra-fibrous biomimetic mineralized collagen membrane capable of promoting osteogenic differentiation of cells

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Experimental program
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Effect test

preparation example Construction

[0037] The specific method for preparing the type I collagen film of biomimetic mineralization is:

[0038] Dissolve the lyophilized powder of calf epidermis in 0.1mol / L acetic acid solution (pH=2.5) to prepare a 3-5g / L collagen solution, store it at 4°C for 24 hours and use it within three months. Pour the collagen solution into a prefabricated mold and put it into a closed container containing 1% ammonia water. After neutralizing for 4 hours, remove it and put it in a 36-38°C incubator to continue culturing for 20 hours. During this process, the collagen completes self-assembly and becomes Gel-like. In order to stabilize the collagen structure, 0.025% glutaraldehyde was used for cross-linking for 1 h. At this point, the pure collagen gel was prepared. Put the pure collagen gel in the biomimetic mineralization solution and culture it at 37°C for 2 days to 4 days respectively to prepare collagen gels with different degrees of biomimetic mineralization, cool down the collagen ...

Embodiment 1

[0040] (1) The concrete method of synthesizing the biomimetic mineralization liquid containing amorphous calcium phosphate is:

[0041] 8g / L polyacrylic acid was used as a stabilizer for amorphous calcium phosphate, and 400mM glutamic acid solution was used as a remineralization accelerator. At 36°C, use 0.1M disodium hydrogen phosphate aqueous solution with a pH of 9.5±0.1 and 0.2M calcium chloride aqueous solution to make mother liquor; use a 50ml syringe to add calcium chloride aqueous solution at a volume ratio of 1:1 within 30 minutes In the disodium hydrogen phosphate aqueous solution, stir with a magnetic stirrer during the reaction process, and use 0.1M sodium hydroxide solution to control the pH to maintain the interval of 9.5±0.5; so that the final concentration of calcium ions in the prepared biomimetic mineralization solution is 9.0mM , the final concentration of phosphorus ions was 5.0 mM, and the final concentration of glutamic acid was 10 mM. The final concentr...

Embodiment 2

[0045] (1) The concrete method of synthesizing the biomimetic mineralization liquid containing amorphous calcium phosphate is:

[0046] 9g / L polyacrylic acid was used as a stabilizer for amorphous calcium phosphate, and 400mM glutamic acid solution was used as a remineralization accelerator. At 37.5°C, use 0.17M disodium hydrogen phosphate aqueous solution with a pH of 9.5±0.1 and 0.28M calcium chloride aqueous solution to make mother liquor; use a 50ml syringe to add calcium chloride aqueous solution at a volume ratio of 1:1 within 37 minutes In the disodium hydrogen phosphate aqueous solution, a magnetic stirrer is used to stir during the reaction, and a 0.1M sodium hydroxide solution is used to control the pH to maintain the interval of 9.5±0.5; so that the final concentration of calcium ions in the prepared biomimetic mineralization solution is 10.0mM , the final concentration of phosphorus ions was 6.3 mM, and the final concentration of glutamic acid was 75 mM. The final...

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Abstract

The invention discloses a preparation method of an intra-fibrous biomimetic mineralized collagen membrane capable of promoting osteogenic differentiation of cells. According to the invention, the preparation method comprises the following steps: preparing a collagen solution; pouring the collagen solution into a prefabricated mold, putting the mold into a closed container containing ammonia water,carrying out neutralizing, moving the neutralized solution out and then putting the solution into a constant-temperature box to carry out culturing continuously, and completing self-assembling and forming a gel mode; carrying out cross-linking on the gel to obtain pure collagen gel; and putting the pure collagen gel into a biomimetic mineralization solution and carrying out culturing for a plurality of days to obtain collagen gel with different biomimetic mineralization degrees; and cooling the collagen gel and carrying out freeze-drying on the cooled gel in a freeze dryer to obtain the finalintra-fibrous biomimetic mineralized collagen membrane. According to the invention, intra-fibrous mineralization of the collagen fiber can be formed by biomimetic mineralization; the fiber network structure of the collagen membrane is reserved and utilized while the mechanical property and the biological property of the hydroxyapatite crystal are utilized.

Description

technical field [0001] The invention relates to a preparation method of a collagen film, in particular to a preparation method of a biomimetic mineralized type I collagen film in fibers that can promote osteogenic differentiation of cells. Background technique [0002] Dental implants have now become a standard and commonly used procedure to restore missing teeth, but the lack of bone in the maxillofacial region due to factors such as trauma, tumors, and periodontal diseases has always been a challenge for oral implants and restorations. Therefore, there are many methods trying to solve this problem, including autologous bone grafting, alveolar ridge splitting and guided bone tissue regeneration (GBR) and so on. Among them, GBR is one of the most commonly used methods and is widely used in anterior implants. Bone substitute materials combined with barrier membranes are required in GBR to block the rapidly proliferating epithelium and connective tissue to ensure osteoblast g...

Claims

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Application Information

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IPC IPC(8): A61L27/12A61L27/24A61L27/50
CPCA61L27/12A61L27/24A61L27/50A61L2430/12
Inventor 顾新华王婧茅彩云陈超群孙健
Owner THE FIRST AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE
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