自体成纤维细胞移植治疗声带瘢痕的实验研究

Experimental study of autologous fibroblast cell transplantation in the treatment of vocal fold scar

目的:研究声带内移植入自体成纤维细胞后声带瘢痕的变化特点。方法:对40只新西兰大白兔以锐性损伤单侧声带制作声带瘢痕模型后,取自体皮肤组织采用酶消化法培养成纤维细胞。将模型分为实验组及对照组,将培养至第3代的成纤维细胞经CM-DIL染料标记后注射入实验组声带瘢痕周围,1周后采用荧光显微镜观察成纤维细胞注射后的存活情况,1个月后行组织学检查,观察声带瘢痕组织的形态改变,并运用RT-PCR方法测定瘢痕声带和移植成纤维细胞声带与正常声带中Ⅰ型、Ⅲ型胶原蛋白的表达差异。结果:自体成纤维细胞移植1周后可在注射声带内观察到荧光标记细胞,移植1个月后,声带瘢痕内胶原蛋白组织结构与正常声带内胶原蛋白组织结构相近,声带瘢痕中Ⅰ型、Ⅲ型胶原蛋白表达比例与正常声带组织中的比例相近。结论:自体成纤维细胞移植入声带后可在声带内存活,接受自体成纤维细胞移植后声带瘢痕可恢复至正常声带结构。

Objective：To study the change characteristics of vocal cords scar after the implantation of autologous fibroblast cells into the vocal cords.Method：Forty New Zealand rabbits were used for the production of vocal fold scar model by creating a unilateral vocal fold wounds,we take the skin tissues of each rabbits and cultured them via enzyme digestion method to obtain autologous fibroblast cells.The models were separated into the experimental group and the control group,the third generation fibroblast cells with CM-DIL labeled were then injected into the surrounding area of the vocal fold scar in the experimental group,the survival situation of the injected cells were observed by fluorescence microscopy after 1week,histological examination was performed after 1months to observe the morphological changes of vocal fold scar tissues,RT-PCR method was used to measure the expression variation of typeⅠcollagen and typeⅢcollagen in the vocal cords and the normal vocal cords,respectively.Result：After 1week of autologous fibroblast transplantation into the vocal folds,the fluorescence labeled cells were observed.1month after transplantation,the collagen protein in the vocal fold scar tissues was similar to that in the normal vocal folds,the expression of typeⅠcollagen and typeⅢ collagen in vocal fold scars is similar to those in the normal vocal folds.Conclusion：Autologous fibroblast migration into the vocal cords can survive well inside the vocal cords,and the vocal scar can be recovered to normal vocal cord structure after transplantation of autologous fibroblast cells.