羊膜负载骨髓间充质干细胞与表皮细胞对放创性皮肤损伤促愈合研究

目的探讨治疗放创性全厚皮肤缺损创面的方法及效果. 方法贵州小香猪8只,每只背部脊柱两侧均有放创性全层皮肤缺损圆形创面(Ф3.67cm)各3个,共48个创面.将经处理的人羊膜(human amniotic mambrane, HAM)分别负载自体骨髓间充质干细胞(mesenchymal stem cells, MSCs)和表皮细胞,移植到其左侧24个创面作为实验组(A组);以单纯无种植细胞的HAM敷盖其右侧前16个创面(B组);以单纯油纱布敷盖其右侧后8个创面(C组).B、C作为对照组.观察移植后1～3周内各组创面愈合、肉芽组织生长及上皮化等情况,并进行创面组织HE染色及vWF免疫组织化学检测.用图像分析法测算各组各时间点创面平均面积(cm2),并计算其愈合百分率. 结果 C组于伤后 22～23天愈合,B组于伤后19～21天愈合;A组于伤后15～17天愈合,较B、C组分别提前6～7天和5～6天,愈合质量好.移植15～17天,A组与B、C组创面平均残留面积及愈合面积百分率比较,差异有统计学意义(P＜0.01). A组创面的新生上皮已完全覆盖整个创面,肉芽组织生长旺盛,肉芽组织中vWF、成纤维细胞和毛细血管含量丰富,可见胶原沉积;B、C组创面仍见许多炎性细胞浸润,肉芽组织中vWF、成纤维细胞和毛细血管含量少,胶原沉积不明显. 结论 HAM负载自体MSCs和表皮细胞植入对放创性全厚皮肤缺损创面有较好的促愈合作用,愈合质量较高.

STUDY ON HUMAN AMNIOTIC MEMBRANE LOADED WITH MARROW MESENCHYMAL STEM CELLS AND EPIDERMIS CELLS IN PROMOTING HEALING OF WOUND COMBINED WITH RADIATION INJURY

OBJECTIVE: To investigate the results of human amniotic membrane (HAM) which are loaded with marrow mesenchymal stem cells (MSCs) and epidermis cells in treating full-thickness skin defect combined with radiation injury. METHODS: Eight minipigs were used in this study. Three round full-thickness wounds (phi3.67cm), which combined with radiation injury, were created on the dorsum of each side close to the vertebral column in each animal. Among 48 wounds, 24 left side wounds were treated with HAM loaded with MSCs and epidermis cells as experimental group (group A), 16 right side wounds with simple HAM (HAM group, group B) and 8 right side wounds with oil gauze as control (group C). The granulation tissue, reepithelization and wound area were observed after 1,2 and 3 weeks. Immunohistochemistry was performed using vWF as a marker for blood vessels. Image analysis was employed to test new area of wound at different interval time and healing rate of wound. RESULTS: The healing time of group A was 6 to 7 days faster than that of group C and 5 to 6 days fister than that of group B. After 15-17 days of graft, there were significant differences in new area of wound and healing rate between group A and groups B, C (P < 0. 01). New epidermis fully covered whole wound surface in group A, and their granulation tissue, which contained a lot of vWF, fibroblasts, capillaries and collagen, grew well. Many inflammatory cells still were seen in groups B and C, and their contents of vWF, fibroblasts, capillaries and collagen in granulation tissue were smaller than that in group A. CONCLUSION: The graft of HAM loaded with MSCs and epidermis cells played an effective role in promoting healing of wound combined radiation injury with high quality.