BMSC And HUVEC-derived Exosome-incorporated Mesoporous Bioglass Scaffold Promote Bone Regeneration By Improving Osteogenesis And Angiogenesis

Objective1?To solve the problems of treatment of large area bone defects caused by trauma,infection,tumor or genetic malformation and promote the regeneration of the missing bone,we characterized a bone regeneration scaffold material,mesoporous bioglass,and studied its conduction in bone(used to induce new bone formation)and bone stimulation(promoting new bone formation).And biological properties in terms of angiogenesis(induced angiogenesis).2?To characterize exosomes and studied their functions in promoting proliferation,adhesion,migration and differentiation in human umbilical vein endothelial cells and bone marrow mesenchymal stem cells by cell experiments.3 ? To study the ability to promote bone regeneration by animal experiments Combining mesoporous bioglass and exosomes.MethodCorresponding studies were performed on HUVEC-Exo and BMSC-Exo,including scanning electron microscopy(SEM),transmission electron microscopy(TEM),NTA,FNA,WB,phagocytosis test LSCM,and extraction HUVEC-Exo and BMSC-Exo were added to HUVEC and BMSC to observe the effect on HUVEC and BMSC osteogenic differentiation,and finally loaded on the MBG scaffold and implanted into the skull defect of rats.After a certain time,the material was taken for MicroCt.scan and perform data statistics to verify the osteogenesis effect.Result1 ? The proliferation,migration,tube formation experiments,VEGF fluorescence,VEGF release and protein expression levels of HUVEC cultured under certain conditions:There was a statistically significant difference between the BMSC-Exo L /BMSC-Exo H group and the Control group?Among them,BMSC-Exo L and BMSCExo H are stronger than Control group..There also was a statistically significant difference between the BMSC-Exo L and BMSC-Exo H groups.Among them BMSC-Exo H is stronger than BMSC-Exo L group.2?Proliferation,migration,tube formation experiments,VEGF fluorescence,VEGF release and OPN,ALP,RUNX-2 gene expression and protein expression levels of BMSCs cultured under certain conditions:There was a statistically significant difference between the HUVEC-Exo L /HUVEC-Exo H group and the Control group?Among them,HUVECExo L and HUVEC-Exo H are stronger than Control group.There also was a statistically significant difference between the HUVEC-Exo L and HUVEC-Exo H groups.Among themHUVEC-Exo H is stronger than HUVEC-Exo L group.3?After the animal model of skull defect was established,cultured for 3 months,MicroCT scan images showed that BMSC-Exo L and BMSC-Exo H were stronger than the Control group,and BMSC-Exo H osteogenesis was stronger than quantitative data.BMSC-Exo L group.ConclusionIn conclusion,we demonstrated in vitro that BMSC-derived exosomes can promote the expression of HUVEC-related genes PI3K/AKT,ERK1/2,promote HUVEC proliferation,migration,osteogenic and VEGF secretion;and HUVEC-derived exosomes It can promote the activation of WMSC,promote the activation of Wnt/?-Catenin pathway,promote the proliferation,migration and VEGF expression of BMSC,and promote the expression of corresponding osteogenic genes BMP-2,OPN,ALP,COL-1 and RUNX-2.Through in vivo experiments,it was preliminarily confirmed by MicroCT that MBG loaded with BMSC-Exo can significantly promote bone defect healing.In summary,mesoporous bioglass-loaded BMSC-derived exosomes promote bone regeneration.