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Double layer artificial skin and preparation method thereof

An artificial skin, double-layer technology, applied in the field of double-layer artificial skin and its preparation, can solve the problems of unfavorable wound edge epidermal cells crawling on the wound surface, difficult to observe the wound infection situation, increase immune rejection and other problems, so as to reduce scar hyperplasia, The effect of preventing wound infection and good biocompatibility

Active Publication Date: 2012-09-19
SHAANXI RUISHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the commercialized double-layer artificial skin has Integra (produced by Integra LifeSciences), which is an artificial skin with a double-layer matrix. It is composed of a silicone membrane, which can control the evaporation of wound water vapor. During the wound healing process, the dermis will combine with the wound. The silicone membrane of the epidermis can be removed after the dermis heals, and then further autologous skin transplantation. The US FDA has approved it in 1996 Integra was approved to be used in the treatment of burns and scalds in 2010; its main disadvantage is that it is difficult to observe the early infection of the wound surface due to the non-degradable adhesive film on its surface, which is not conducive to the crawling of wound epidermal cells and the sealing of the wound surface, and the silicone film is removed later Second surgery is needed
Apligraf (Organogenesis company production) is also the artificial skin of double layer, and dermis is planted in bovine collagen by human dermal fibroblast, and epidermis is made of human epidermal cells, is attached to form epidermis on the dermis; Its deficiency mainly is The epidermis is composed of allogeneic epidermal cells with high immunogenicity, which increases the immune rejection of the product, and at the same time makes the production process complicated, long in production time, and increased in cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Step 1). Acquisition of fibroblasts: Take the foreskin tissue after neonatal resection, wash it with phosphate buffer saline (PBS) after disinfection, remove the subcutaneous fat and muscle layer, cut the skin into strips, and use 4U / ml medium Protease solution for digestion for 2 hours; remove the epidermis, cut the dermal tissue into pieces, digest with 625U / ml collagenase solution for 2.5 hours, and harvest fibroblasts for culture; use cell factory or spinner bottle for the primary cultured fibroblasts Cultivation systems, or methods such as bioreactors to expand cultivation.

[0020] Step 2). Preparation of decellularized small intestinal submucosa: Rinse fresh porcine jejunum with distilled water, cut into required segments, soak and sterilize in an aqueous solution containing 0.2% peracetic acid and 10% ethanol for 1 hour, and rinse with PBS solution Rinse; mechanically scrape off the small intestinal mucosa, muscle layer and serosa, rinse with PBS to obtain the s...

Embodiment 2

[0025] Step 1). Acquisition of fibroblasts: refer to the literature Dermal matrix as a carrier for in vivo delivery of human adipose-derived stem cells. Biomaterials. 2008Apr; 29(10): 1431-42. It has been proved that adipose-derived mesenchymal stem cells can differentiate for fibroblasts. Take healthy adipose tissue discarded during the operation, wash it with phosphate buffered saline (PBS) after disinfection, cut up the adipose tissue, digest it with 625U / ml collagenase solution for 150 minutes, and inoculate the harvested adipose-derived mesenchymal stem cells into culture bottles Cultivate; and then use the method of cell factory, or spinner bottle culture system, or bioreactor to expand the culture.

[0026] Step 2). Decellularized dermis preparation: rinse the fresh pig skin with distilled water, remove the epidermis and subcutaneous tissue layer mechanically, cut into 0.4mm thick, 5cm 2 Size, soak and disinfect in an aqueous solution containing 0.2% (v / v) peracetic ac...

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Abstract

The invention discloses a double-layer artificial skin and a preparation method thereof, wherein, cell-free membranous biological derivative material is used as a surface layer, and a fibroblast, extracellular matrix synthesized and secreted by the fibroblast and a cell growth factor are compounded in the interior of biological support material to form a dermis, and then the surface layer and thedermis are combined to form the double-layer artificial skin; a compact surface layer structure can effectively reduce the loss of water, electrolytes and protein from surface of wound, avoid the invading and the reproduction of bacteria to the impaired surface of wound as well as the infection of the surface of wound, and be beneficial to epitheliosis and epithelial growth; the dermis can directly repair the surface of wound, promote the ingrowth of cells around the surface of wound and the angiogenesis, induce the differentiation from stem cells to skin cells and quicken wound healing; the artificial skin has the advantages of being capable of promoting the regeneration of skin, improving the elasticity, the flexibility and the mechanical abrasion resistance of skin after the surface ofwound is healed, reducing excess scar tissues, controlling the contracture, having excellent biocompatibility, increasing the success rate of transplant and improving the quality of healing; the double-layer artificial skin has wide material resources and simple production method, and is applicable to the clinical treatment of skin defect caused by inflammation, ulcer, thermal burns, iatrogenicity and the like.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering biomaterials, and in particular relates to a double-layer artificial skin and a preparation method thereof. Background technique [0002] Skin is an important tissue organ that covers and protects the body surface. Defects and abnormalities of the skin are often caused by inflammation, ulcers, trauma, burns, postoperative tumors, and congenital deformities. For this kind of tissue defect, the method of autologous skin transplantation is mostly used at present, which not only causes new trauma defects in the donor area, but also is often limited by the source of skin donor. [0003] With the development of tissue engineering technology, the preparation of artificial skin has become a hot spot in the treatment of skin defects. Ideal tissue-engineered skin can imitate the structure of natural skin, with a composite skin of epidermis and dermis. The composition of the dermis is close to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/60A61L27/38
Inventor 王爱军
Owner SHAANXI RUISHENG BIOTECH
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