壳多糖-胶原-糖胺聚糖凝胶人工皮肤的初步研究  

PRIMARY STUDY ON THE CHITOSAN-COLLAGEN-GAGS COLLAGEN GEL ARTIFICIAL SKIN

作　　者：朱堂友[1] 伍津津[1] 胡浪[1] 李文维[1] 贺萍[1] 张民众[2] 徐春蓉[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所皮肤科,重庆400042 [2]重庆医科大学儿童医院

出　　处：《中国修复重建外科杂志》2003年第2期113-116,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目 (39870 2 38)~~

摘　　要：目的　研制一种新型的胶原凝胶类人工皮肤。方法　制备壳多糖 -胶原 -糖胺聚糖 (GAGs) -成纤维细胞真皮替代物 (DE) ,观察成纤维细胞 (FB)在凝胶中的生长情况和不同因素对凝胶收缩的影响 ,并绘制 FB生长曲线和凝胶收缩曲线。了解不同含量壳多糖对 FB和角质形成细胞 (KC)生长的影响 ,不同含量壳多糖 DE的抗感染能力。再于“成熟”的 DE表面接种 KC,先浸没培养 ,后行气液界面培养 ,构建完整的人工皮肤。对 DE和人工皮肤行组织学和电镜分析。结果　 DE收缩度与 FB数量呈正比 ,终末收缩度与胶原蛋白浓度成反比 ;FB在凝胶中 2～ 9天呈指数增生。 DE基质配方对 FB的生长无明显抑制作用 ,但可促进 KC的生长 ,对金黄色葡萄球菌的抑制作用随壳多糖含量增大而增强。扫描电镜示 DE有丰富的微孔结构。人工皮肤组织学观察见类似于正常皮肤 ,有分化良好的表皮和致密的真皮。结论壳多糖 -胶原 - GAGs胶原凝胶人工皮肤是一种新型、有一定抗感染能力的活人工皮肤。Objective To constitute a new collagen gel artificial skin by using chitosan as one of the components. Methods Human foreskin fibroblasts were incorporated into the chitosan collagen GAGs to constitute dermal equivalent(DE). The growth of fibroblasts incorporated in gels and several factors which influenced the contraction of the gel were observed. The influence of different chitosan contents on the growth of fibroblast and keratinocyte and on the antibacterial effect were studied. Keratinocytes separated from normal children foreskin were seeded on the matured DE to reconstruct artificial skin, which was immersed at the early stage of culture, then lifted to an air liquid interface. The structure of the DE and artificial skin were analysed by histology and scanning electron microscope. Results The contraction rate of the DE was proportional to the number of fibroblasts, and the final size of the DE was inversely proportional to the concentration of collagen protein. Fibroblasts incorporated into the gel showed the exponential growth from the 2nd day to the 9th day. Chitosan collagen GAGs had no inhibition effect on the growth of fibroblasts, but promoted the growth of keratinocytes. Staphylococcus aureus was inhibited even more as chitosan content increased. Scanning electron microscopy indicated that the DE had abundant porous fabrication. Artificial skin shared some histological features of normal skin, which consisted of a good stratified epidermis and a dense dermis. Conclusion Chitosan Collagen GAGs collagen gel artificial skin is a new collagen gel living artificial skin which has certain antibacterial ability and stratified epidermis and dense dermis structure like normal skin.

关 键 词：壳多糖-胶原-糖胺聚糖 人工皮肤 胶原凝胶类 真皮替代物 

分 类 号：R318.19[医药卫生—生物医学工程]