VEGF165 对脱细胞真皮基质血管化影响的研究

Effects of VEGF165 on vascularization of acellular dermal matrix

关键词:  脱细胞真皮基质  血管内皮生长因子  血管化

Key words:  acellular dermal matrix  vascular endothelial growth factor  vascularization

摘要:

［目的］观察十二烷基硫酸钠（sodium dodecyl sulfate，SDS）和NaOH复合消蚀法制备的胎儿脱细胞真皮基质（ADM）的组织相容性，以及血管内皮生长因子（VEGF）对ADM血管化的影响。［方法］ 将足月胎儿皮肤用SDS和NaOH复合消蚀,制备ADM（厚0.3 mm）。用重约300 g的SD雄性大鼠45只，随机分为ADM、ADM+rAAV/LacZ 和ADM+rAAV/VEGF₁₆₅3组进行真皮片皮下包埋实验。术后的2、4、6周分别取材，经HE染色或以VEGF165、基质金属蛋白酶-1（MMP-1）、组织型基质金属蛋白酶抑制剂-1（TIMP-1）的单克隆抗体进行免疫组织化学染色，检测3种蛋白质在移植后的皮肤中的表达情况。观察并比较各组中ADM血管密度，进行图像处理和统计学分析。［结果］ADM无细胞成分，胶原纤维结构规整；术后2周包埋ADM组织中可见到新生血管，及VEGF阳性细胞；无明显的炎性细胞浸润。ADM临近组织中MMP-1和TIMP-1均明显表达，MMP-1的表达高于TIMP-1；术后4周各组细胞数量和新生血管均高于2周。此时MMP-1的表达高于TIMP-1，但TIMP-1的表达较2周时有所增强；术后6周rAAV/VEGF₁₆₅实验组中VEGF阳性细胞和血管密度明显高于ADM空白组和rAAV/LacZ对照组（P<0.05）。此时TIMP-1的表达则高于MMP-1。［结论］SDS和NaOH复合消蚀法制备的ADM去除细胞彻底，并保持了真皮中原有细胞外基质构筑的完整，而且免疫原性低，组织相容性好；VEGF₁₆₅与ADM复合移植，具有促进ADM血管化作用；MMP-1和TIMP-1的相互作用参与植入体内的ADM的血管化过程。

Abstract:

［Objective］To observe the biocompatibility of acellular dermal matrix(ADM) prepared by SDS and NaOH maceration,and VEGF₁₆₅ on the effects of the vascularization of ADM.［Methods］Randomly 45 mice were devided into three groups-ADM vacnity group,ADM+rAAV/LacZ control group and ADM+rAAV/VEGF165 experiment group.ADMs were embed between skins and skin-muscles on the backs of mice. 2,4,6 weeks after operations mice skins were drawn the materials from back section. Then they were characterized with HE staining and immunohistochemical staining to detect the retention of VEGF₁₆₅,MMP-1 and TIMP-1 staining. To examine the quantities of them in the ADMs,morphometric and statistical analyses were performed.［Results］The cellular components of ADM were virtually absent and the form and arrangement of its collagen fibers had not been changed obviously with HE staining; Two weeks after operation there were a few cells and new capillaries in ADMs tissue. At this time ,MMP-1 and TIMP-1 obviously expressed in tissues and the expressions of MMP-1 were more than TIMP-1. Four weeks after operation, in every group, the quantities of cells and new capillaries, were more than 2 weeks. At this time, the expression of MMP-1 increased, and the expression of TIMP-1 also increased. Six weeks after operation, the quantities of cells and new vessels had obviously altered. In ADM+ rAAV/VEGF₁₆₅ experiment group, the quantities of cells expressing VEGF₁₆₅ and vessels density were obviously higher than the others (P<0.05). The expression of TIMP-1 was higher than MMP-1.［Conclusions］No cell components is observed in the ADM prepared by the maceration of SDS and NaOH. The architectural integrity of the dermal extracellar matrix is not damaged;The compound transplantation of VEGF₁₆₅ and ADM can promote the vascularization of ADM;MMP-1 and TIMP-1 take part in the vascularization of ADM.