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Artificial skin containing human bone marrow mesenchymal stem cell and its construction method

A bone marrow mesenchymal and artificial skin technology, applied in the field of artificial skin, can solve the problems of toxic hazards in the body, no artificial skin, strong immune antigenicity, etc., achieve good flexibility and enzyme resistance, and induce autologous cell growth , accelerated healing effect

Inactive Publication Date: 2004-07-14
RADIOLOGY INST ACAD OF MILITARY MEDICINE SCI PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The U.S. patent of US 777419 provides a method for constructing artificial composite skin using bovine type I and III collagen, but the artificial skin has the following disadvantages: 1. Glutaraldehyde is used as a cross-linking agent, which is toxic to the body and has a low strength. Not high; 2. Unremoved collagen epitopes, strong immunogenicity; 3. Using pure type I and III collagen as a scaffold without adding epidermal growth factor
The U.S. patent of US 5800811 provides a method for constructing artificial skin using a collagen matrix containing transforming growth factor, but the patent has the following disadvantages: 1. The artificial skin is only a collagen scaffold without adding cell components; 2. The collagen antigen is not removed to determine 3. Although it has been proved that this scaffold can combine with bone marrow mesenchymal stem cells, artificial skin with bone marrow mesenchymal stem cells growing on both sides has not been constructed.
The US patent of US 6391059 discloses the technology of implanting bone marrow mesenchymal stem cells in the artificial skin scaffold for repairing bone defects, but the scaffold components are simple and have not been double-layer implanted

Method used

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  • Artificial skin containing human bone marrow mesenchymal stem cell and its construction method
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Examples

Experimental program
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Effect test

Embodiment 1

[0035]The fetal cowhide is depilated, fascia, fat and blood stains are removed, cleaned and frozen according to the conventional method, cut into 0.5mm thin slices, dehydrated with acetone three times, degreased with petroleum ether three times, and then vacuumed to remove petroleum ether to make dehydrated degreasing Fetal calf leather is used as a raw material. 10 g of the raw material is taken, washed with double distilled water, and then added to the reactor, and an aqueous solution prepared from 0.1 g of gastric enzyme and 100 g of water is added. Carry out an enzyme treatment, at 20°C, pH=2, mechanically shake for 24 hours to degrade the covalent bonds between tissue fibers, centrifuge, discard the supernatant, take the precipitate, wash three times with double distilled water, add 500g 0.5M acetic acid aqueous solution, at 0°C, mechanically vibrate for 32 hours, then add sodium chloride to make the concentration of sodium chloride in the solution reach 2M, after the prec...

Embodiment 2

[0039] The bovine Achilles tendon is dehydrated, degreased, and vacuumized to remove petroleum ether according to the method in Example 1, and the dehydrated and degreased bovine Achilles tendon is made as a raw material. Take 10 g of the raw material, wash it with double distilled water, and add it to the reactor. Add 0.3 1 g of trypsin and 100 g of the prepared aqueous solution were subjected to an enzyme treatment. Under the conditions of 30 ° C and pH = 6, mechanical vibration was performed for 16 hours to degrade the covalent bonds between tissue fibers. Wash with distilled water for four times, add 1000g 0.5M acetic acid aqueous solution, mechanically vibrate for 28 hours at 10°C, then add sodium chloride to make the concentration of sodium chloride in the solution reach 2M, after the precipitation is complete, centrifuge and discard the supernatant , take the precipitate, put it into the dialysis unit and carry out dialysis four times, collect the collagen solution after...

Embodiment 3

[0043] Use the bovine Achilles tendon dehydrated and degreased in Example 2 as raw material, get 10g, add in the reaction vessel after washing with double distilled water, add the aqueous solution that is prepared by 0.5g pineapple enzyme and 100g water and carry out enzymatic treatment once, at 40 ℃, pH Mechanical vibration under the condition of =8 for 8 hours degrades the covalent bonds between tissue fibers, centrifuges, discards the supernatant, takes the precipitate, washes it four times with double distilled water, adds 1500 g of 0.5M acetic acid aqueous solution, and dissolves it at 20°C After 24 hours of mechanical vibration, add sodium chloride to make the concentration of sodium chloride in the solution reach 2M. After the precipitation is complete, centrifuge, discard the supernatant, take the precipitate and put it into a dialysis bag for dialysis three times, collect the dialysis collagen solution and add In the reactor, add 0.1g chymotrypsin and 100g water to pre...

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Abstract

The present invention relates to construction process of artificial skin containing human bone marrow mesenchymal stem cell. Skin tissue engineering rack is first constructed with fetus calf or ox tendo achillis as basic material, collagen sponge film through twice enzyme treatments and the mixture of heparin and epidermal growth factor; and human bone marrow mesenchymal stem cell obtained via purifying human bone marrow blood is implanted on two sides of the rack to constitute the artificial skin with bone marrow mesenchymal stem cell on the two sides of and in the internal of the rack. The artificial skin has no antigenicity, high strength, high flexibility, no toxicity to wound, and capacity of inducing wound cell growth and promoting wound healing.

Description

technical field [0001] The invention relates to an artificial skin containing human bone marrow mesenchymal stem cells, which belongs to a medical skin wound face covering. [0002] The invention also relates to the construction method of the artificial skin. Background technique [0003] In addition to hematopoietic stem cells, human bone marrow tissue also contains a certain amount of bone marrow mesenchymal stem cells. Bone marrow mesenchymal stem cells are relatively immature cells that have the potential to differentiate into epidermal and dermal tissues and promote wound healing. [0004] In vitro cell culture of bone marrow mesenchymal stem cells to the 10th passage, the cells still have differentiation potential, which lays the foundation for the use of cultured and proliferated bone marrow mesenchymal stem cells to construct artificial skin. [0005] Skin wound coverings include epidermal grafts, dermal grafts, and composite skin grafts. The U.S. patent of US 7774...

Claims

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Application Information

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IPC IPC(8): A61F2/02A61L27/60C12N5/00
Inventor 刘杰张燕崔雪梅王德文杨颖颙于淑贤
Owner RADIOLOGY INST ACAD OF MILITARY MEDICINE SCI PLA
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