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Preparing method of heterogeneous acellular dermal matrix substrate with good biocompatibility

A decellularized dermis and biocompatibility technology, applied in the field of preparation of heterogeneous acellular dermal matrix, can solve the problems of crosslinking agent toxicity, uncontrollable, unstable cytotoxicity, etc., to reduce the dielectric constant and improve tissue phase Capacitance, the effect of protecting physical properties

Active Publication Date: 2015-11-25
HEBEI AINENG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at the same time of cross-linking, more importantly, the toxicity of the cross-linking agent itself and its residue become a hidden cause affecting the cytocompatibility
3. In addition, in the process of our industrialization research, we found that a major problem of products prepared by decellularization technology is that the cytotoxicity is unstable and difficult to control
Domestic marketed products, uncrosslinked and crosslinked tissue engineering materials still have cytotoxic substandard products, although they have been washed with phosphate buffer and purified water in the preparation process of decellularization technology, they still have cytotoxicity

Method used

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  • Preparing method of heterogeneous acellular dermal matrix substrate with good biocompatibility
  • Preparing method of heterogeneous acellular dermal matrix substrate with good biocompatibility
  • Preparing method of heterogeneous acellular dermal matrix substrate with good biocompatibility

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] (1) Preparation of tomographic skin slices

[0069] Take 0.6mm thick porcine split skin slices, cut into 5*7cm size, wash with PBS three times, remove fat with 0.1% Promojieer for 30 minutes, then wash with PBS three times;

[0070] (2) Separation of epidermis and dermis

[0071] Soak the split skin piece obtained in step (1) with 0.2% (mass volume ratio g / 100ml) trypsin in PBS solution at 20°C for 16 hours, wash with 70% ethanol for 8 hours, and wash with 3% hydrogen peroxide for 15 minutes, then Wash with PBS for 3 times, then wash with purified water for 24 times, and change the medium every 1 hour;

[0072] (3) Decellularization treatment

[0073] Wash the material after step (2) with 0.3% (mass volume ratio g / 100ml) SDS aqueous solution for 4 hours, wash with 4% ethanol for 4 hours, continue to wash with purified water for 12 times, change the medium every 1 hour, and finally PBS Wash with liquid 24 times, change the liquid every two hours;

[0074] (4) Cross-l...

Embodiment 2

[0085] (1) Preparation of tomographic skin slices

[0086] Take a 0.6mm thick sectioned animal skin slice, cut it into 5*7cm size, wash it three times with PBS solution, remove fat with 0.1% Promojieer for 30 minutes, and then wash it three times with PBS solution;

[0087] (2) Separation of epidermis and dermis

[0088] Use 0.15% (mass volume ratio g / 100ml) trypsin in PBS solution to soak the split skin piece obtained in step (1) at 4°C for 30 hours, wash with 40% ethanol for 10 hours, and wash with 3% hydrogen peroxide for 15 minutes. Then wash 3 times with PBS, then wash 24 times with purified water, and change the medium every 1 hour;

[0089] (3) Decellularization treatment

[0090] Wash the material after step (2) with 0.1% (mass volume ratio g / 100ml) SDS aqueous solution for 6 hours, 2% ethanol for 6 hours, continue to wash with purified water for 12 times, change the medium every 1 hour, and finally PBS Wash with liquid 24 times, change the liquid every two hours; ...

Embodiment 3

[0101] (1) Preparation of tomographic skin slices

[0102] Take a 0.6mm thick sectioned animal skin slice, cut it into 5*7cm, wash it three times with PBS solution, remove fat with 0.1% Promogeril for 30 minutes, and wash it three times with PBS solution;

[0103] (2) Separation of epidermis and dermis

[0104] Use 0.25% (mass volume ratio g / 100ml) of trypsin in PBS solution obtained in step (1), soak for 24 hours at 10°C, wash with 75% ethanol for 6 hours, and wash with 3% hydrogen peroxide for 15 minutes. Then wash 3 times with PBS, then wash 24 times with purified water, and change the medium every 1 hour;

[0105] (3) Decellularization treatment

[0106] Wash the material after step (2) with 0.5% (mass volume ratio g / 100ml) SDS aqueous solution for 3 hours, wash with 10% ethanol for 2 hours, continue to wash with purified water for 12 times, change the medium every 1 hour, and finally PBS Wash with liquid 24 times, change the liquid every two hours;

[0107] (4) Cross-...

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Abstract

The invention relates to a preparing method of a heterogeneous acellular dermal matrix substrate with good biocompatibility. The method concretely comprises the following steps of (1) split thickness skin graft preparation; (2) epidermis and dermis separation; (3) acellular processing; (4) cross linking; (5) virus inactivation; (6) refining; (7) Co-60 irradiation sterilization. The ecellular tissue engineering materials obtained by the method provided by the invention has the advantages that the toxicity is stable, and the cell compatibility is better.

Description

Technical field [0001] The present invention involves the field of organizational engineering and specifically involves a preparation method for the dermis matrix of alien dehydration in a good biocompatibility. Background technique [0002] The development of modern biomedicine pays great attention to cell -based studies, including the composition and change of alignment of intracellular biomass.On the one hand, the research on molecular levels is valued, and on the other hand, the relationship between cells, organs, and the completion of the body is also a very important research content.Such awareness prompts people to pay more attention to the substances of mechanical support and physical intensity for tissue, organs, and even the integrity of the entire body in the lower level of the epithelium or endothelial cells, and the integrity of the entire body.In recent years, research on the types, structures, functions, and regulation of extracellular matrix has become very import...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
Inventor 宋灵杰李一礼李晶谢美娜王云泽杨国峰
Owner HEBEI AINENG BIOTECH CO LTD
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