自体表皮细胞悬液移植修复全层皮肤缺损创面的动物实验研究

发布时间：2018-11-22 12:14

【摘要】：目的探讨自体表皮细胞悬液移植技术用于全层皮肤缺损创面修复的适宜密度。方法取健康清洁级成年SD大鼠40只,雌雄不限,体质量210~230 g;根据细胞移植密度不同,随机分为高、中、低细胞密度及空白组(分别为A、B、C、D组,n=10)。取大鼠背部皮肤培养表皮细胞,并制作大鼠全层皮肤缺损创面抗挛缩模型。其中A、B、C组分别将0.2 mL密度为1×10~6、1×10~5、1×10~4个/cm~2的自体表皮细胞悬液移植至创面处,D组给予等量限制性角质形成细胞无血清培养基;取成年Wistar大鼠背部皮肤制备同种异体皮,覆盖各组创面。术后观察大鼠存活情况,于术后7、14、21 d大体观察同种异体皮成活、脱落及创面愈合情况,同种异体皮脱落后计算创面愈合率;21 d时取材行组织学及免疫组织化学染色,观察创面修复情况。结果术后大鼠均存活至实验完成。各组大鼠同种异体皮随时间延长逐渐成活,干燥并开始脱痂;至21 d同种异体皮基本脱落后A、B组创面可见成片上皮,C组创面可见少量菲薄上皮,D组创面无上皮形成。术后21 d同种异体皮脱落后,A、B、C、D组创面愈合率分别为62.9%±9.6%、64.2%±9.1%、38.5%±5.7%、22.7%±5.5%,A、B组创面愈合率显著高于C、D组(P0.05),C组高于D组(P0.05),A、B组间比较差异无统计学意义(P0.05)。组织学观察示,A、B、C组愈合的创面上皮层可见鳞状上皮细胞,A、B组表皮分层明显,C组表皮层薄、可见炎性细胞浸润,D组为肉芽组织。免疫组织化学染色观察示,A、B、C组表皮-真皮连接层Ⅳ型胶原和Ⅶ型胶原表达呈阳性,D组无表皮层呈阴性;A、B组Ⅳ、Ⅶ型胶原表达阳性细胞百分比显著高于C组(P0.05),A、B组间比较差异无统计学意义(P0.05)。结论自体表皮细胞悬液移植技术在大鼠全层皮肤缺损创面修复中可重构皮肤,1.0×10~5个/mL为创面修复的适宜移植密度。
[Abstract]:Objective to investigate the proper density of autologous epidermal cell suspension transplantation for full-scale skin defect repair. Methods Forty healthy clean grade adult SD rats, male and female, 210g / 230g, were randomly divided into high, middle and low cell density and blank group (n = 10). The skin of rat back was cultured with epidermal cells and the model of anti contracture was established. In group A, the suspension of autologous epidermal cells with a density of 0. 2 mL 1 脳 10 ~ 6 and 1 脳 10 ~ 5 / cm~2 was transplanted to the wound, and group D was given the same amount of serum-free medium for restricted keratinocytes. Allogeneic skin was prepared from the dorsal skin of adult Wistar rats to cover the wounds in each group. The survival of the rats was observed after 714 days. The survival rate of allogeneic skin and wound healing were observed on the 21st day after operation. The rate of wound healing was calculated after allogeneic skin exfoliation. The wound repair was observed by histological and immunohistochemical staining at 21 d. Results all the rats survived until the experiment was completed. The allogeneic skin in each group survived gradually with time, dried and began to unscab, and at 21 days after the abscission of the allogeneic skin, the lamellar epithelium was found in group A B, a small amount of thin epithelium was found in group C, and no epithelium was formed in group D. On the 21st day after operation, the healing rate of the wounds in the group A BX Con D was 62.9% 卤9.6% and 64.2% 卤9.1% respectively, and 38.5% 卤5.7% 卤5.5A, respectively, in the group A, the healing rate of the wounds was 22.7% 卤5.5A, and the healing rate of the wounds in group A was 62.9% 卤9.6% 卤9.2% 卤9.1%, respectively. The rate of wound healing in group B was significantly higher than that in group C (P0.05) (P0.05) in group), C and group D (P0.05), but there was no significant difference between group B and group A (P0.05). Histological observation showed that squamous epithelium cells could be seen in the epithelium of wound healing in group A BX C, epidermis in group A B was obviously stratified, epidermis in group C was thin and inflammatory cells were infiltrated, and granulation tissue was found in group D. Immunohistochemical staining showed that the expressions of type 鈪，

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