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A kind of preparation method of human-derived acellular dermal matrix

A decellularized dermis and decellularized technology, which is applied in the field of medical materials, can solve the problems of complicated process, complicated steps, and increased environmental pollution, and achieve the effects of small immune rejection, simple and reliable production process, and reduced environmental pollution

Active Publication Date: 2021-08-17
上海仁康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many preparation methods of acellular dermal matrix, there are more or less certain problems: the current preparation methods of acellular dermal matrix are very complicated, need enzyme digestion, lye treatment, etc., the steps are complicated and the process is complicated, which greatly increases the cost. Production cost; most of the current preparation methods of acellular dermal matrix use chemical fixatives, such as glutaraldehyde, which greatly changes the natural structure of acellular dermal matrix and hinders the acellular dermal matrix in vivo Degradation; At the same time, glutaraldehyde residues cause greater cytotoxicity and genotoxicity of decellularized products, which greatly affects clinical use; At present, there are also many acellular dermal matrix preparation methods that introduce alkaline solution and organic solvent treatment, which not only affects the decellularized products. The structure of the dermal matrix also increases the possibility of environmental pollution, and the treatment of waste liquid also increases the cost; such as repeated freezing and thawing, this method belongs to the use of physical methods for decellularization, which takes a long time and a long cycle. no longer needed

Method used

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  • A kind of preparation method of human-derived acellular dermal matrix
  • A kind of preparation method of human-derived acellular dermal matrix
  • A kind of preparation method of human-derived acellular dermal matrix

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] A preparation method of human-derived acellular dermal matrix, the specific steps are as follows:

[0038] (1) Take the stored human skin;

[0039] (2) Remove subcutaneous fat components;

[0040] (3) Step (2) removes the subcutaneous skin with fatty components, cleans and disinfects with 2% iodophor, deiodines with 75% alcohol, and then repeatedly soaks and cleans the skin with sterile distilled water;

[0041] (4) Treat the skin after step (3) washing with 2.5mg / ml Dispase II solution overnight;

[0042] (5) Discard the Dispase II solution, remove the epidermis, and then wash with physiological saline to obtain the dermis;

[0043] (6) the dermis that step (5) obtains contains Ca with 2mol / L 2+ NaCl solution for 18h, Ca 2+ The final concentration is 5mM / L;

[0044] (7) After discarding the 2mol / L sodium chloride solution, wash with normal saline;

[0045] (8) Place the dermis cleaned in step (7) in the prepared "decellularized solution", which contains the followi...

Embodiment 2

[0048] Example 2 Internal Structure of Acellular Dermal Matrix

[0049] (1) Cut the acellular dermal matrix obtained above into a size of 1 cm × 1 cm, and paste it on a metal tray; the section of the acellular dermal matrix is ​​quenched with liquid nitrogen to make it hard and brittle, and then broken; the section faces upward Adhesive to the metal tray.

[0050] (2) After spraying gold in the sample preparation room of the electron microscope, it was put on the machine, observed and photographed by the scanning electron microscope, and the results were as follows: image 3 shown.

[0051] The results showed that there were no cell residues in the tissue, and the collagen structure was complete, similar to normal skin.

Embodiment 3

[0052] Example 3 HE staining of acellular dermal matrix

[0053] (1) Fix the normal skin tissue and the acellular dermal matrix in 4% paraformaldehyde fixative solution for 24 hours respectively;

[0054] (2) Alcohol dehydration, xylene transparency;

[0055] (3) Dip in wax for 30 minutes, embed, and make sagittal paraffin sections with a thickness of 5 μm;

[0056] (4) Gradient alcohol carries out dewaxing, dehydration, HE staining;

[0057] (5) Transparent, sealed, observed under a light microscope, the results of HE staining of normal skin tissue are as follows: Figure 4 As shown, the results of HE staining of acellular dermal matrix are as follows Figure 5 shown.

[0058] The results showed that there were no cell residues in the skin tissue.

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Abstract

The invention discloses a preparation method of a human-sourced acellular dermal matrix. The specific steps are as follows: take the stored human-sourced skin; remove subcutaneous fat components; clean and disinfect with iodophor and alcohol, and then repeatedly soak and clean with a hypotonic solution Skin; treated overnight with Dispase II solution; removed epidermis, and then washed with normal saline to obtain dermis; treated with hypertonic saline solution; washed with normal saline; treated with decellularized solution in a dialysis device to obtain decellularized dermal matrix; Wash with normal saline to obtain human-derived acellular dermal matrix. The preparation method of the present invention is simple, the production process is simple and reliable, the production efficiency is improved, the production cost is saved, and the environmental pollution can be reduced to the greatest extent. At the same time, the obtained acellular dermal matrix retains the skin basement membrane, which is more conducive to cell adhesion and proliferation. Rich in growth factors, it has good biocompatibility and is more conducive to supporting the growth of cells.

Description

technical field [0001] The invention relates to the technical field of medical materials, in particular to a method for preparing human-derived acellular dermal matrix. Background technique [0002] Dermal substitutes play an important role in the wound healing process. They can provide a dermal scaffold at the bottom of the wound, improve the survival rate of the cultured epidermal membrane, and reduce wound shrinkage and scar formation. Skin grafts are the best treatment for patients with extensive burns or trauma. Acellular dermal matrix (ADM) has an internal structure similar to that of skin dermis, and has been used as a dermis substitute in both domestic and foreign experiments and clinics. [0003] Currently, there are two main types of acellular dermal matrix: allogeneic acellular dermal matrix and heterogeneous acellular dermal matrix. Compared with the heterogeneous acellular dermal matrix, practice and clinical use have confirmed that the allogeneic acellular de...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/60
CPCA61L27/362A61L27/3687A61L27/60
Inventor 董亚兵朱宁文朱迎春
Owner 上海仁康科技有限公司
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