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Formula and preparation method for gel support used for in-vitro construction of tissue-engineered cartilage

A tissue engineering, gel-like technology, applied in the field of cartilage tissue engineering scaffold and its preparation, can solve the problem of secondary injury of patients

Active Publication Date: 2015-12-30
科医联创(浙江)生物科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, the source of chondrocytes is generally taken from the patient's own cartilage in the non-weight-bearing area. The more chondrocytes are needed, the more cartilage needs to be taken, which will cause secondary damage to the patient. For large-area cartilage defects, cartilage tissue engineering There is a good curative effect, but due to the above reasons, although the number of treatments for articular cartilage is huge, the existing feasible cell therapy methods cannot effectively meet this demand

Method used

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  • Formula and preparation method for gel support used for in-vitro construction of tissue-engineered cartilage
  • Formula and preparation method for gel support used for in-vitro construction of tissue-engineered cartilage

Examples

Experimental program
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Embodiment 1

[0025] This example provides a gel-like scaffold formulation for constructing tissue engineered cartilage in vitro, including gel and additives, wherein the gel includes fibrin 100mg / ml, collagen 25mg / ml, hyaluronic acid 5%, thrombin 50U / ml ml; the final concentration of additives in DMEM / F12 (1:1) basal medium is:

[0026] Vitamin C: 233μM~287μM, Linoleic acid: 3.7μM~8.9μM, Cholesterol: 9μM~17μM, Dexamethasone: 6nM~15nM, Acetylcysteine: 43μM~58μM, Transferrin: 18μg / mL~32μg / mL, Sodium Selenite: 22nM~52nM, Sodium Pantothenate: 13μM~24μM, Biotin: 28μM~43μM, Insulin: 7μg / mL~18μg / mL, Epidermal Growth Factor: 2ng / mL~9ng / mL, Fibroblasts Growth factor: 2ng / mL~9ng / mL, platelet-derived growth factor: 2ng / mL~9ng / mL, human serum albumin 0.5%~2.5%.

[0027] When fibrin and thrombin are mixed, the last step of the coagulation chain reaction is simulated, and the fibrinogen is gradually polymerized through the activation of thrombin on fibrinogen, and finally a fibrin solidified product i...

Embodiment 2

[0029] This embodiment provides a preparation method of a gel-like scaffold for constructing tissue engineered cartilage in vitro, including:

[0030] 1) The vitamin C, linoleic acid, cholesterol, dexamethasone, acetylcysteine, transferrin, sodium selenite, sodium pantothenate, biotin, insulin, epidermal growth factor, fibroblast growth factor, platelet derived Growth factors, human serum albumin, were added to the basal medium DMEM / F12 so that the final concentration of the additive in the basal medium was:

[0031] Vitamin C: 233μM~287μM, Linoleic acid: 3.7μM~8.9μM, Cholesterol: 9μM~17μM, Dexamethasone: 6nM~15nM, Acetylcysteine: 43μM~58μM, Transferrin: 18μg / mL~32μg / mL, Sodium Selenite: 22nM~52nM, Sodium Pantothenate: 13μM~24μM, Biotin: 28μM~43μM, Insulin: 7μg / mL~18μg / mL, Epidermal Growth Factor: 2ng / mL~9ng / mL, Fibroblasts Growth factor: 2ng / mL~9ng / mL, platelet-derived growth factor: 2ng / mL~9ng / mL, human serum albumin 0.5%~2.5%;

[0032] 2) Dissolve and dilute fibrin, coll...

Embodiment 3

[0034]Additives: Vitamin C, Linoleic Acid, Cholesterol, Dexamethasone, Acetyl Cysteine, Transferrin, Sodium Selenite, Sodium Pantothenate, Biotin, Insulin, Epidermal Growth Factor, Fibroblast Growth Factor, Platelet Derivative Growth factors, human serum albumin were dissolved in the basal medium DMEM / F12 (1:1) in proportion; so that the final concentrations of each supplementary component in the basal medium were: vitamin C: 250 μM, linoleic acid: 4.5 μM , Cholesterol: 13μM, Dexamethasone: 10nM, Acetylcysteine: 50μM, Transferrin: 25μg / mL, Sodium Selenite: 30nM, Sodium Pantothenate: 17μM, Biotin: 33μM, Insulin: 10μg / mL, Epidermal growth factor: 5ng / mL, fibroblast growth factor: 5ng / mL, platelet-derived growth factor: 5ng / mL, human serum albumin 1%.

[0035] Dissolve and dilute fibrin, collagen, hyaluronic acid and thrombin with the solution prepared in the previous step to make the final concentration of fibrin 100mg / ml; collagen 25mg / ml; hyaluronic acid 5%; thrombin 50U / ml ....

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Abstract

The invention provides a formula and a preparation method for a gel support used for in-vitro construction of tissue-engineered cartilage. The formula comprises gel and an additive, wherein the gel may be fibrin, collagen, hyaluronic acid and thrombin, and the additive may be vitamin C, linoleic acid, cholesterol, dexamethasone, transferrin, biotin and the like. With the formula and the support prepared by using the method, cartilage cells cultured in vitro maintain activity of original cells; and the gel support with plasticity can be provided according to the shape of a cartilage defect of a patient in clinical application of cartilage tissue engineering.

Description

technical field [0001] The invention relates to the field of cartilage repair, in particular to a cartilage tissue engineering scaffold and a preparation method thereof. Background technique [0002] Articular cartilage damage, whether caused by trauma or pathological causes, cannot be repaired spontaneously, and usually leads to degeneration of cartilage, resulting in osteoarthritis. At present, the application of cartilage tissue engineering technology can treat such traumatic (or osteochondritis exfoliative) knee articular cartilage defects. arthroscopic equipment, and used in cartilage tissue engineering technology. [0003] For example: Chinese Patent Publication No. CN103877615B discloses a cartilage tissue engineering scaffold and a preparation method thereof. The method includes preparing a demineralized bone matrix, preparing a natural polymer hydrogel solution, and mixing the natural polymer hydrogel solution with the Injection into the decalcified bone matrix, s...

Claims

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Application Information

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IPC IPC(8): A61L27/26A61L27/20A61L27/22A61L27/24A61L27/54
Inventor 柴勋吴国祥李文琳袁彩艳韩武梅张易
Owner 科医联创(浙江)生物科技集团有限公司