Reconstruction Studies On Tissue Engineering Artificial Skin With Porous Modified Polylactic Acid Scaffold.

Title: Reconstruction Studies on Tissue Engineering Artificial Skin With ModifiedPolylactic Acid Scaffold.Tissue engineering is the product of co-development and combination of modern cell biology,biomaterial science, and engineering. The aim of tissue engineering is to investigate and restoretissue and organ substitutes. In this study, we use PLA which is no-toxic, no-hot, biocompatibilityand biodegradability , and modified PLA as scaffold construct tissue engineering corium and tissueengineering artificial skin. Details as following:Ⅰ. Isolation, cultures and subcultures of KC and FB.KC isolated from new born rat by treatment of trypsin-dispase was cultured,subcultured, and adopted with techniques of containing serum . It not only isolated theepidermal and dermal, but also obtained all cell of basal stratum. The method pre-digest with trypsin and the method pre-digest with EDTA are thebest of methods in the subculturing KC.Both of them could avoide the damage of KCcoming from trypsin and advanced the efficiency of subculturing.KC grows very wellafter subcultured.FB was isolated and cultured by the normal methods. According to the principles of plantingwith high cell density(1.0×105cell/ml) and refeed in short time established, the FB was purifiedafter subcultures.Ⅱ. Effects of some kinds of vitamin and coenzyme on the proliferation of KC and FBThe effects of some kinds of vitamin and coenzyme on KC and FB proliferation were studiedby means of the MTT -method. The results showed that VC,CoA, ATP had the promotional effectson proliferation of KC. VB6 had the restraining effects on proliferation of KC. VB6, VC, CoA, ATPhad the promotional effects on proliferation of FB.Ⅲ Biocompatibility of five different modified PLA scaffold and reconstruction of tissueengineering artificial coriumA kind of method that salt out scaffold preparation is used to make scaffold.Scaffold is made from PLA combined with linear alkyl benzene sulfonate(LAS),dialdehyde starch, polyvinylalcohol (PVA) and glycerin. The artificial corium was reconstructed by planting subcultured FB separatelyinto(onto) five kinds of modified Polylactic Acid sponge, cultured for two week. Theresults were studied by means of the MTT -method. There are four kinds of modifiedpolylactic acid sponge witch FB have grown very well except LAS.Ⅵ Reconstruction of tissue engineering artificial skin.The artificial skin was reconstructed by planting subcultured FB and KCseparately into(onto) the sponge, cultured for one week.The artificial skin wascomposed of epidermis and dermis under light microscope, the skin is consisted of KCand FB .KC and FB have grown very well in the scaffold.In all,the research bases on the successful isolation and culture of KC and FB,andstudies the best subculture method of KC and the effect of some kinds of vitamine andcoenzyme on the proliferation of KC and FB.What's more,it explores thebiocompatibility of cells with the modofied PLA ( PLA-DAS , PLA-PVA ,PLA-glycerin) and PLA scaffold.The result shows that FB grow well in the scaffoldand it convinced that the scaffold could be used for new scaffold material of artificialskin.