Research Of The Effects And Related Machenisms Of VH298 On Promoting Bone And Soft Tissue Defects Repairment

Objective: Hypoxia inducible factor(HIF)has been shown to promote a variety of physiological functions of cells.This research focused on three difficult issues in reconstruction surgery,bone defect,refractory wound and enthesis rupture.In order to solve these problems,VH298,an agonist of hypoxia inducible factor,was used to explore its therapeutic potential and related mechanisms.Methods: In the cell experiments,Western blot was used to determine the expression of hypoxia inducible factor in different cells after VH298 treatment,and the time-effect and concentration-effect curves were established,which established the basis for subsequent experiments.In this study,CCK-8 assay was used to determine the proliferation activity of different cells,scratch assay was used to evaluate the migration ability of cells,and fluorescence quantitative PCR was used to detect the expression of cell function-related genes.The differentiation potential of stem cells was detected by inducing differentiation to different cell lines.The tube forming experiment was used to detect the angiogenesis ability of vascular endothelial cells in vitro.Different concentrations of VH298 were used to in these experiments.Compared with control group,the best effective concentration of VH298 in vitro was determined.In the aspect of animal experiments,the rat model of distraction osteogenesis,diabetic wound model and Achilles tendon-calcaneal enthesis reconstruction model were established to simulate clinical cases.Then the optimal concentration of VH298 determined in the cell experiment was used to intervene the healing process of the three animal models.The effects of VH298 on the healing of bone and soft tissue defects in animal models were evaluated by imaging findings,biomechanical experiments and histological staining,and the experimental phenomena in vivo were explained by the results of cell experiments.Results: It was found that the concentration of intracellular hypoxia inducible factor increased with the elevating of VH298 concentration and reached the highest value after 200 ?M VH298 treatment;with the prolongation of culture time,the concentration of intracellular hypoxia inducible factor generally reached the peak at 2 hours of culture and then slowly declined.Subsequent studies found that 30 ?M VH298 generally had the best effect of promoting cell proliferation,migration,differentiation and functional gene expression.After local application of 30 ?M VH298,the mechanical efficiency and callus formation in the distraction osteogenesis area were significantly improved,and the microvascular formation in diabetic wounds was more vigorous and showed obvious acceleration of healing process.The reconstructed tendon-bone interface showed more mature and stronger mechanical tensile properties.Conclusion: appropriate concentration of VH298 can effectively promote the mineralization of new bone in distraction area and accelerate the healing of diabetic wound and tendon-bone interface.This shows that VH298 has a strong application potential facing of the same clinical problems.