Enoxaparin Interferes With The Process Of Osteogenesis In Vivo And Vitro Studies

Objective:1.To study the effect of enoxaparin on differentiation and osteogenesis of bone marrow mesenchymal stem cells into osteoblasts;2.To study the effect of enoxaparin on bone healing process after bone defect;3.To study the characteristics of exosome derived from bone marrow mesenchymal stem cells under the intervention of enoxaparin.Methods:1.Extract the bone marrow mesenchymal stem cells from SD rats,build vitro model of osteogenesis under the intervention of enoxaparin,using CCK 8 experimental detection for cell inhibition rate,flow cytometry instrument detection for cell cycle and apoptosis,kits for ALP activity,PCR for the expression of OCN,BMP2 and VEGF genes,Western Blot for the expression of the proteins,alizarin red staining for calcium nodules;2.Build rat femur bone defect model under the intervention of enoxaparin,using X-ray imaging system for rat model successful building and bone repair,Micro CT for bone repair related data analysis,immunohistochemical staining for protein expression of OCN and TRAP;3.Extract the exosomes from the bone marrow mesenchymal stem cells with the intervention of enoxaparin,using transmission electron microscopy to observe the morphology of exosomes,and using Elisa to detect the osteogenic related proteins in exosomes.Results:1.Enoxaparin has certain inhibitory effect for osteogenesis,on one hand inhibits cell proliferation and promotes cell apoptosis,on the other hand suppresses expression of osteogenesis related genes and proteins;2.High concentration of enoxaparin(2000AXa IU/ml)has a certain inhibitory effect on bone defect repair;3.The content of proteins OCN and BMP2 in the exosomes derived from bone marrow mesenchymal stem cells under the intervention of enoxaparin was significantly higher.Conclusion:Enoxaparin has a certain inhibitory effect on the osteogenesis process,and the exosomes information transmission may be the negative regulation mechanism of this process.