Double-layered artificial skin and its prepn process

A technology of artificial skin and dermis, applied in the field of medicine and biomedical engineering, can solve the problems of difficult operation, large shrinkage rate of collagen gel, unpredictable immune rejection, etc.

Inactive Publication Date: 2004-01-21
上海组织工程研究与开发中心
View PDF0 Cites 16 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of Apligraft is that the collagen gel has a large shrinkage rate; using allogeneic cells and bovine co

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Double-layered artificial skin and its prepn process
  • Double-layered artificial skin and its prepn process
  • Double-layered artificial skin and its prepn process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Isolation and culture of keratinocytes

[0059] 1) Take materials

[0060] The surgically removed foreskin of children is placed in the keratinocyte culture medium containing antibiotics and brought into the cell culture room.

[0061] 2) Separate the epidermis

[0062] Carefully cut off the subcutaneous fat tissue and part of the dermis with scissors, and trim them into strips with a width of about 1-2mm, rinse twice with calcium and magnesium-free PBS, and soak in 0.24U / ml neutral protease (Dispase II) with the epidermis facing upward overnight at 4°C.

[0063] 3) digestion

[0064] After 18 hours, the epidermis was torn off from the dermis with two ophthalmic tweezers, cut into pieces, and then digested with 0.05% trypsin-0.53mM EDTA at 37°C for 15 minutes, and 10ml trypsin inhibitor (10mg / ml) to stop the digestion . Filter through a 150-mesh stainless steel filter, centrifuge at 1000r / min for 10 minutes, discard the supernatant, add 10ml of keratinocyte culture...

Embodiment 2

[0070] Isolation and culture of dermal fibroblasts

[0071] 1) digestion

[0072] Cut the dermis after peeling off the epidermis into pieces, digest with 0.1% collagenase at 37°C for 2 hours, filter through a 150-mesh stainless steel filter, centrifuge at 1000r / min for 5 minutes, discard the supernatant, and add calcium-free magnesium Wash 3 times with PBS. Add 10ml of DMEM culture solution containing 10% fetal bovine serum, mix well, and trypan blue staining and counting.

[0073] 2) Primary culture

[0074] Make a single cell suspension after counting, press 1-2×10 6 / 75cm 2 Density inoculated culture, DMEM medium containing 10% fetal bovine serum, at 37°C, 5% CO 2 , cultured in an incubator with 100% relative humidity, and the culture medium was changed every 3 days.

[0075] 3) Subculture

[0076] When the cells are confluent to 60%-75% density, wash with 10ml calcium and magnesium-free PBS, digest with 0.25% trypsin at 37°C for 5 minutes, add DMEM culture medium co...

Embodiment 3

[0078] Preparation of artificial skin

[0079] (1) Construction of fibroblast-PGA complex

[0080] The dermal fibroblasts prepared in Example 2 were collected by trypsin digestion, and then were divided into 0.2 × 10 6 / 10mg polyglycolic acid, 1×10 6 / 10mg polyglycolic acid, 5×10 6 It was inoculated on a network of polyglycolic acid (PGA) at a concentration of 10 mg polyglycolic acid. Then inoculate the inoculated PGA material at 37±0.5°C, 5% CO 21. Under the condition of saturated humidity, culture in DMEM culture solution containing 10% fetal bovine serum (the culture solution is changed every 3 days) for 1-2 weeks to form fibroblast-PGA complex.

[0081] (2) Formation of double skin

[0082] The keratinocytes prepared in Example 1 were collected, and then respectively divided into 0.5×10 5 / square centimeter, 5×10 5 / square centimeter, 50×10 5 / cm 2 seeded on the surface of the fibroblast-PGA complex prepared in the previous step. The inoculated complex was mixed ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Thicknessaaaaaaaaaa
Thicknessaaaaaaaaaa
Login to view more

Abstract

The present invention provides one kind of double-layered artificial skin implant comprising one hypodermal layer containing polyglycollic acid and other biodegradable material, fibroblast and matrix protein; and one epidermal layer on the hypodermal layer. The artificial skin has continuous epidermal cell growing on the surface of the fibroblast-PGA composite, well cell differentiation and obvious lamination. The slice shows the artificial skin has the tissue structure characteristic similar to that of normal body skin. The preparation process and use of the artificial skin is also provided.

Description

technical field [0001] The invention relates to the fields of medicine and biomedical engineering, and more specifically relates to a double-layer artificial skin constructed with polyglycolic acid as a scaffold, and a preparation method and application thereof. Background of the invention [0002] Skin covers the whole body surface of the human body and is in direct contact with the external environment. It is an important boundary organ in anatomy and physiology. Skin accounts for about 16% of adult body weight, with an area of ​​about 1.2-2.2m 2 . The skin is composed of epidermis and dermis, connected with deep fascia, aponeurosis or periosteum by subcutaneous tissue. The skin originates from the ectoderm and mesoderm, and has a complex and highly specialized structure. It plays an important barrier and protective role, preventing external stimuli from damaging tissues in the body, blocking the invasion of foreign bodies and microorganisms, and preventing the extravasa...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61L27/60
Inventor 曹谊林杨光辉崔磊刘伟
Owner 上海组织工程研究与开发中心
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap