胶原-磺化羧甲基壳聚糖人工真皮支架修复Ⅲ度烧伤创面时组织TGF-β1表达与细胞凋亡的研究  

Repair of full-thickness burn wound with collagen-sulfonated carboxymethyl chitosan dermal stent in pigs

作　　者：徐少骏[1] 马列[2] 黄爱宾[2] 滕建英[3] 朱金土[1] 倪有娣[3] 叶盛[3] 徐奂军[3] 王永光[3] 

机构地区：[1]浙江中医药大学附属第一医院整形外科,杭州310006 [2]浙江大学材料与化学工程学院 [3]杭州师范大学临床医学院

出　　处：《浙江医学》2010年第5期640-643,I0001,共5页Zhejiang Medical Journal

基　　金：浙江省医药卫生科学研究基金A类（2008A131）

摘　　要：目的 探讨胶原-磺化羧甲基壳聚糖人工真皮支架修复Ⅲ度烧伤创面时生物合成替代和细胞凋亡的机制,为临床应用提供理论依据.方法 将胶原-磺化羧甲基壳聚糖/硅橡胶双层人工皮肤支架移植于10头猪Ⅲ度烧伤清创后创面,对植入后1、2、3周的创面及植入后2周创面加植表皮后2周的修复情况进行观察.同时,通过免疫组织化学、TUNEL、天狼猩红染色方法,对不同时间的创面组织中转化生长因子p 1（TGF-p 1）的表达水平、细胞凋亡发生情况以及支架自身胶原替代情况进行观测.以不植入人工真皮支架的同期烧伤创面作为对照.结果 （1）实验组植入人工真皮支架后2～3周,创面颜色逐渐红润,并变光滑,而对照创面表面粗糙;（2）实验组TGF-β 1表达水平在人工真皮支架植入后1～2周持续增高且高于对照组,3～4周持续下降且低于对照组.对照组1～3周的创面持续升高,4周下降;（3）实验组植入后2～4周凋亡细胞持续增多且数量显著高于相应对照组,对照组3～4周时凋亡细胞持续增多;（4）人工真皮支架植入后1周,自身胶原已开始合成,植入后3周基本完成支架自身胶原替代.结论 胶原-磺化羧甲基壳聚糖人工真皮支架在创面修复中有明显作用,其修复创面的机制与自然的肉芽或瘢痕修复不同,在创面修复重建中有良好的应用前景.Objective To investigate the application of collagen-sulfonated carboxymethyl chitosan porous scaffold transplantation in repair of full-thickness burn wound in animal model. Methods The wound surface was observed at 1, 2, and 3 weeks after the bilayer dermal equivalent （BDE） composed of collagen-chitosan porous scaffold and silicone membrane were transplanted on wounds of full thickness burn with debridement in 10 Bama miniature pigs, and so was done the wound healing after epidermis had been grafted for 2 weeks on surface of the scaffold transplanted on wounds for 2 weeks. At same time, TGF-β1 expressions, apoptosis and self collagen replacement of scaffolds in burn wounds were detected by immunohistochemistry, TUNEL method and picrosirius red staining, respectively. Wounds without scaffold transplantation were studied as control. Results Wounds of the scaffold transplantation became ruddy and smooth from 3 to 4 weeks after BDE transplantation, which were different from granulation wounds. TGF- β1 expressions in the transplantation wounds increased continuously from 1 to 2 weeks and were higher than those in the control wounds, and decreased continuously from 3 to 4 weeks and were lower than those in the control wounds after BDE transplantation. TGF-β1 expressions in the control wounds increased continuously from 1 to 3 weeks and decreased in 4 weeks. Apoptosis signals increased continuously in the transplantation wounds from 2 to 4 weeks after BDE transplantation and were significantly higher than those in the corresponding control wounds, whereas, signals increased continuously in the control wounds from 3 to 4 weeks. Self collagen was synthesized in the transplantation wounds at 1st week and scaffolds was replaced by self collagen at 3rd week after BDE transplantation by observation with picrosirius red polarized light method. Conclusion Collagen-sulfonated carboxymethyl chitosan plays an important role in wound healing of full thickness burn. The mechanism of wound repair by dermal scaffold is diffe

关 键 词：双层人工皮肤支架 胶原-磺化羧甲基壳聚糖 Ⅲ度烧伤 创面修复 TGF-Β1 细胞凋亡 天狼猩红 

分 类 号：R692.202[医药卫生—泌尿科学]