The Effect Of Recombination Angiogenesis Inhibitor Ad-METH-1 On Hypertrophic Scar

To investigate the relation between angiogenesis and hypertophic scar formation thoroughly and study the effect of anti-angiogenesis on hypertophic scar by injecting Ad-METH-1 into excessive scar on ears of rabbit.First, the microvessels in hypertrophic scar at different clinical phase were observed by H&E staining and immunohistochemistry staining, meanwhile, the microvessels in excessive scar on ears of rabbits were studied by microcirculation microscope and H&E staining, the microcirculation in tissue of scar were detected by Laser Doppler. The result showed that the microvessel density in tissue of scar at proliferative phase was significantly higher than that at mature period (P<0.05). It is similar to hypertrophic scar on ears of rabbits (P<0.05). Furthermore, the microcirculation appears resemble regularity as to the capillary in scar. The results hint us that angiogenesis enough at the early proliferative phase might have an important influence on formation of hypertrophic scar, and it could be depressed effectively by anti-angiogenesis at the early period of hypertrophic scar.In the following test, the recombinated endostatin YH-16 was injected into hypertrophic scar on ears of rabbits at the tenth day after re-epithelization,then the effect of YH-16 on hypertrophic scar was observed by microcirculation microscope. Laser Dopple, H&E staining and TEM( transmission electron microscope ). It is found out that angiogenesis and tissue proliferation were depressed markedly after injection of YH-16. It proved that hypertrophic scaring could be effectively held back by anti-angiogenesis.Second, adenovirus particle with pAdEasy-methl were injected into the early scar of animal model at the tenth day after re-epithelization, then the effect of anti-angiogenesis by gene transfection on hypertrophic scar was studied. At the thirtieth day after injection, the specimens were collected, then the proliferation of fibroblast was analyzed by AgNOR staining, the synthesis of I and III type collagen was study by collagen staining and in situ hybridization of I and III type procollagen mRNA, at the same time, the micro vessels in scar were observed by CD34 immunohistochemistry staining. The results showed that angiogenesis, proliferation of fibroblast and synthesis of I and III type collagen in tussue of scar were depressed after Ad-METH-1 being injected (P<0.05). It is proved Ad-METH-1 could effectively repress the formation of hypertrophic scar by anti-angiogenesis in the early phase after re-epithelization.To investigate the mechanism of Ad-METH-1 on hypertrophic scar, in the sequent test, the effect of Ad-METH-1 on cultural HUVEC was researched. Cell activity and cell cycle were studied by MTT and FCM at the twentieth hour after Ad-METH-1 being added into cultural HUVEC (human umbilical vein endothelial cells), in the following step, ET-1 and bFGF in supernatant were detected by RIA (radioimmunoassay) and ELASA (enzyme-linked immuno assay). The result showed that Ad-METH-1 depressed effectively...