不同人工真皮支架在修复Ⅲ度烧伤创面中创面收缩和细胞凋亡差异的实验研究

Differences of wound contraction and apoptosis in full-thickness burn wounds repaired with different artificial dermal stent in pigs

目的 探讨不同真皮支架血管化对Ⅲ度烧伤创而修复中创面收缩和细胞凋亡影响的差异.方法 将胶原-磺化羧甲基壳聚糖、胶原-壳聚糖及脱细胞基质三种真皮支架分别移植于猪Ⅲ度烧伤清创后创面,观察植入创面的修复情况,并通过免疫组化、末端脱氧核苷酸介导的生物索化的脱氧尿嘧啶DNA切口末端标记等方法对不同时间小同支架创面中表达α-平滑肌肌动蛋白（α-SMA）的血管和肌成纤维细胞数量及细胞凋亡情况进行检测.结果 植入不同真皮支架的创面与无支架植入的肉芽创面不同.支架植入创面1～3周α-SMA表达阳性的血管数持续增加,支架植入2周加植表皮2周后创面血管数减少,不同时间点以胶原-磺化羧甲基壳聚糖真皮支架植入创面最多,无支架植入创面最少.α-SMA表达阳性的肌成纤维细胞以胶原-磺化羧甲基壳聚糖真皮支架植入创面最少且表达高峰为2周,其他创面表达高峰为3周,以无支架植入创面最多.不同支架植入后2～4周,创面内细胞凋亡持续大幅增多;而无支架植入创面中,3～4周才开始增加;细胞凋亡以胶原-磺化羧甲基壳聚糖真皮支架创面最多,无支架创面最少.结论胶原-磺化羧甲基壳聚糖真皮支架可增强修复细胞的迁移,获得良好的血管化,较好较快地修复皮肤全层烧伤缺损创面.

Objective To investigate the roles and differences of angiogenesis of different dermal scaffolds on wound contraction and apoptosis during full-thickness burn wound repair.Methods Wounds were observed at different time after the collagen-sulfonated carboxymethyl chitosan porous scaffold or collagen-chitosan porous scaffold or acellular dermal matrix were respectively transplanted on wounds of full thickness burn with debriment in Bama miniature pigs.At the same time.vessels and myo-fibroblasts expressing α-smooth musde action（α-SMA）and apoptosis in wounds of different time were detected in situ by immunohistochemical staining.terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphatebiotin nick end labeling,The burn wounds without any scaffold transplantation were studied as the control.Results Wounds with different scaffolds transplantation were different from granulation wounds.Vessels expressing α-SMA had been increasing continuously in the wounds from 1 to 3 weeks after different scaffolds transplantation and decreased in wounds after epidermis had been grafted for 2 weeks on surface of the scaffolds transplanted on wounds for 2 weeks.Vessels expressing α-SMA were the most in the wounds with collagen-sulfonated carboxymethyl chitosan porous scaffold transplantation and the least in the control wounds without dermal scaffold at different time.Myo-fibroblasts expressing α-SMA was the least in the wounds with collagen-sulfonated carboxymethyl chitosan porous scaffold transplantation and the peak of expressions was on the 2nd week,however,the peak in the wounds with the other two scaffolds transplantation and in the control wound without dermal scaffold was on the 3rd week.Myo-fibroblasts expressing α-SMA was the most in the control wounds.Apoptosis had been increasing continuously in the transplantation wounds from 2 to 4 weeks after different scaffolds transplantation,however,apoptosis had begun to increase continuously from 3 to 4 weeks in the control wounds.Apoptosis wag the most in the wounds with collagen-sulfonated carboxymethyl chitosan porous scaffold transplantation and the least in the control wounds without dermal scaffold from 3 to 4 weeks.Conclusion Sulfonated earboxymethyl chitosan can promote migration of reparative cells and angiogenesis,and it can repair full-thickness burn wound fast and well.