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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  • 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

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

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0037] The specific method for preparing the biomimetic mineralized type I collagen membrane is:

[0038] The lyophilized calf epidermis powder was dissolved in a 0.1mol / L acetic acid solution (pH=2.5) to prepare a 3-5g / L collagen solution, stored at 4°C for 24h, and it was effective within three months. Pour the collagen solution into the prefabricated mold and put it into a closed container containing 1% ammonia water. After neutralizing for 4 hours, remove it and place it in a 36~38℃ incubator to continue culturing for 20 hours. In this process, the collagen completes self-assembly and becomes Gel like. In order to stabilize the collagen structure, cross-linked with 0.025% glutaraldehyde for 1 hour, the preparation of the pure collagen gel is completed. Put the pure collagen gel in a biomimetic mineralization solution at 37°C for 2 days to 4 days to prepare collagen gels with different biomimetic mineralization degrees. The collagen gel is gradually cooled to -60°C and placed...

Example Embodiment

[0039] Example 1:

[0040] (1) The specific method for synthesizing the biomimetic mineralized liquid containing amorphous calcium phosphate is:

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

Example Embodiment

[0044] Example 2:

[0045] (1) The specific method for synthesizing the biomimetic mineralized liquid containing amorphous calcium phosphate is:

[0046] 9g / L polyacrylic acid was used as a stabilizer for amorphous calcium phosphate, and a 400mM glutamic acid solution was used as a remineralization accelerator. At 37.5℃, use 0.17M, pH 9.5±0.1 disodium hydrogen phosphate aqueous solution and 0.28M calcium chloride aqueous solution to make the mother liquor; use a 50ml syringe to add the calcium chloride aqueous solution in a volume ratio of 1:1 within 37 minutes In the aqueous solution of disodium hydrogen phosphate, use a magnetic stirrer to stir during the reaction, and use a 0.1M sodium hydroxide solution to adjust the pH to maintain the pH in the range 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 is 6.3mM, and the final concentration of glutamic acid is 75mM....

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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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