Experimental Study On The Vascular Regeneration Of Tissue Engineered Constructs Regulated By MiR126

Objective:Early vascularization after implantation of tissue engineering materials into the body is a key issue in the successful restoration of alveolar bone defects.The formation of new blood vessels inside the material depends on the stimulation of the mobilization and recruitment of endothelial progenitor cells by various angioblast growth factors.Although studies have found that the delivery of growth factors promotes the enhancement of angiogenic capacity in the body,this exogenous direct intervention breaks the balance of revascularization,leading to the formation of a large number of immature blood vessels,which leads to adverse consequences,such as microvascular leakage.MicroRNAs can simultaneously act on multiple target genes and regulate the production of various angiogenic factors to induce regeneration of mature blood vessels.This study was designed to screen for the appropriate microRNAs and to detect the regulation of angiogenesis-related genes by target microRNAs,as well as the effects of angiogenesis and maturation of tissue engineered materials in vivo and in vitro,thus contributing to the successful defect of large-scale alveolar bone.The study could provide new ideas and a theoretical basis for the application of this technology to clinical therapeutic transformation applications.Methods:The miRNA(miR-126)which promotes the expression of the angiogenic phase gene was screened in our previous study.The miRNA(miR-126)which promotes the expression of the angiogenic phase gene was screened by real time RT-PCR and Weston blot.Verify that the target miR-126 is effective in promoting the expression of angiogenic growth factors.Real time RT-PCR and Weston blot were used to verify the effect of miR-126 in promoting the expression of angiogenic growth factors.The experiment was divided into two groups,A and B(experimental group: miR-126+ HUVEC;control group: negative-miRNA control + HUVEC).The ability of miR-126 in promoting angiogenesis in vitro and in vivo was verified by lumen formation experiments and ectopic angiogenesis models in vitro.Transfected HUVEC cells were subcutaneously injected into nude mice to verify the angiogenesis ability of miRNA in vivo.Results:RT-PCR and Weston blot showed that miR-126 can promote the expression of various angiogenic genes and was selected as the target gene(miR-126).We found that miR-126 over-expression could enhance the HUVECs tube formation ability in vitro and stable micro-vessel formation in vivo.Conclusions:MiR-126 over expression could simultaneously enhance multiple angiogenic key factors expression and further induce angiogenesis and vascularization both in vitro and in vivo.These provide evidence to put micro RNA-based therapy into practical clinics as a kind of treatment.