骨髓间充质干细胞自体移植提高猪皮肤创面修复质量的初步研究

目的　探讨骨髓间充质干细胞 (MSCs)局部移植对提高猪皮肤烫伤创面组织修复质量的作用 ,为临床皮肤损伤后功能性修复提供新的治疗方法。方法　抽取 6只小型香猪的骨髓 ,经体外培养并纯化MSCs,应用 5 溴脱氧尿嘧啶 (BrdU)标记技术进行标记。猪背部皮肤中线两侧各制备 6个面积为 2 5 4cm2 的深Ⅱ0 烫伤创面 ,将已标记的MSCs( 2× 10 6)以注射方式回植到提供骨髓猪的创面下 ,将创面随机分为 6组 ,即空白对照组、MSCs治疗组、MSCs 碱性成纤维细胞生长因子 (bFGF)治疗组、bFGF治疗组、MSCs 表皮细胞生长因子 (EGF)治疗组以及EGF治疗组。分别于伤后 7、14、2 1、4 2d采用大体观察、常规组织学检查、免疫组织化学及免疫荧光化学染色动态观察创面愈合情况。结果　实验猪皮肤烫伤后 7d开始 ,各组创面逐渐缩小 ,伤后 2 1d ,大部分创面愈合。虽然不同时间点各治疗组创面面积缩小率差异无显著意义 ,但以MSCs bFGF治疗组最为明显 ,伤后第 14天和 2 1天创面缩小的面积比其他 5组大 15 %～ 2 0 %。半定量评价结果显示MSCs bFGF治疗组肉芽组织中血管密度较大 ,为 ,而其他组别仅为 ～ 。再上皮化的新生表皮在MSCs bFGF治疗组较其余 5组厚 ,并有上皮角形成。半定量分析可见神经纤维在MSCs bFGF治疗组较其他 5

Enhancing the repair quality of skin injury on porcine after autografting with the bone marrow mesenchymal stem cells

OBJECTIVE: To explore the effects of bone marrow mesenchymal stem cells (MSCs) on the quality of healing of porcine skin wounds after burn injury so as to provide a new method for clinical skin repair in the future. METHODS: Seventy-two deep-partial thickness burn wounds were produced on the back of 6 minipigs and then the pigs were randomly divided into 6 groups: saline control, MSCs treatment, MSCs plus basic fibroblast growth factor (bFGF) treatment, MSCs plus epidermal growth factor (EGF) treatment, bFGF treatment or EGF treatment only. MSCs were isolated from porcine marrow and cultured in vitro. After labeling with BrdU, MSCs were autografted onto the skin wounds. 7, 14, 21 and 42 days after the injury, the area of the wounds were measured and the histological examination was performed to evaluate the velocity and quality of wound healing.1, 2 and 4 weeks after transplantation, immunohistochemical examinations were carried out to detect the positive staining of BrdU, cytokeratin and S-100 to evaluate the wound healing quality. RESULTS: The area of wounds was decreased at day 7 and most of these wounds were healed on day 21 after injury. Although there was no significant statistical difference in the contraction rate among these six groups, however, the reduced wound areas in the MSCs plus bFGF treatment group was 15% to 20% more than that in the other 5 groups. Histological examination and semi-quantitative analysis demonstrated that the number of vessels and the expression density of S-100 in the MSCs plus bFGF treatment wounds were significantly enhanced than that in the other groups. CONCLUSION: MSCs autografting may benefit to enhance the wound healing quality in porcine skin, which may open a new way to reach a "perfect repair" after skin injury.