携带HGF基因的脐带血干细胞异种移植促进烧伤创面修复的实验研究

Effects of Ad- HGF modified umbilical cord blood stem cell on healing of burn wounds after xenograft

目的探讨携带人肝细胞生长因子基因的重组腺病毒(Ad-HGF)修饰的脐血干细胞局部异种移植对烧伤创面愈合的作用。方法用密度梯度离心法检测体外分离培养人脐血干细胞,并以感染复数(MOI=100)体外转染Ad-HGF。将24只Wistar大鼠分为脐血干细胞治疗组(A组,n=8),Ad-HGF修饰脐血干细胞治疗组(B组,n=8),空白对照组(C组,n=8)。建立大鼠深Ⅱ度烧伤模型,根据不同组别将细胞悬液及生理盐水于创面下多点注射,伤后3、5、7、14、21 d观察创面收缩率并采集创面标本,行HE染色以及创面组织羟脯氨酸的测定。结果伤后7、14、21 d,B组创口收缩率高于与其他2组,差异有统计学意义意义(P<0.05);HE染色显示,3、5 d各组HE染色差异无统计学意义,7 d可见B组表皮化明显;21 d B组再生表皮明显厚于其它各组,并可见较多的新生血管和皮肤附属器生成。伤后5 d B组羟脯氨酸升高明显,与其他2组比较差异有统计学意义(P<0.05);14 d羟脯氨酸含量降低,与其他2组比较差异有统计学意义(P<0.05)。结论 Ad-HGF修饰脐血干细胞异体移植可显著提高烧伤创面愈合速度和质量,为烧伤创面修复提供了新的策略。

Objective To explore effects of umbilical cord blood stem cell with transfection of adenoviral vector expressing hu- man hepatocyte growth factor （Ad - HGF） after xenograft on healing of burn wounds. Methods Umbilical cord blood stem cell were collected,cultured and transfected with Ad - HGF. 24 Wistar rats were subjected to boiling water for 12 seconds to produce a partial thickness skin burn. The rats were randomly divided into 3 groups：umbilical cord blood stem cell treatment （group A）, transplantation of Ad - HGF modified umbilical cord blood stem cell （ group B）, and saline control （ group C）. According to different groups, the suspen- sion of stem cells, and saline were injected into the derma of burn wounds. The contraction rate of bum wounds was calculated and histo- logical examination was performed at 3,5,7,14 and 21 days after the injury. At the same time, hydroxyproline content in wound tissue was analyzed. Results During the first,second and third week after burn, the contraction rate of burn wounds in group B was much higher than that in group A and C （P 〈 O. 05 ）. Histolgical examination showed the thickness of epidermis in group B was significantly deeper than that in the other groups, and many small vessels were observed in the derma of bum wounds. Hydroxyproline contents of wound tis- sues in group B was higher than others after 5 days, and decreased markedly at the 14th day. Conclusion Modification of umbilical cord blood stem cell with Ad - HGF transfection has positive effects on the healing of burn wounds, which may provide a new way to burn wound repair.