Research On Osteogenic Differentiation Of EXOs-ADSCs-SF-CS Three-dimensional Scaffold Complex In Vitro

Background: Bone regeneration is a complicated process which dependent on the cells,angiogenesis and growth factors work together.The bone tissue engineering is composed of seed cells,scaffold materials and growth factors.Firstly,the ideal seed cells should have multiple differentiation potential,strong regeneration ability and stable nature.Secondly,the structure and mechanical properties of artificial bone scaffold must be excellent.In addition,many experiments in recent years have shown that intercellular signaling is also important for bone regeneration.The compound of adipose-derived stem cells,Silk fibroin-chitosan 3D scaffold and exosomes is expected to provide new ideas for solving this problem.Objective: To investigate the osteogenesis effect of exosomes on the compound of adipose-derived stem cells(ADSCs)and Silk fibroin-chitosan 3D scaffold(SF-CS)in vitro.Methods:1.Extraction,cultivation and identification ADSCs in vitro.2.Using freeze-drying method to prepare the 3D scaffolds.3.ADSCs were induced into osteoblasts and exosomes were extracted by ultracentrifugation.4.ADSCs were seeded on scaffolds and observed by scanning electron microscope.5.The cell-scaffold complex were divided into four groups.We can assay the osteogenic effect of exosomes on ADSCs by detection of osteogenic relate proteins Runx2 and the ALP and CCK-8activity.Results: Compared with the control group,addition of exosomes can significantly enhance the osteogenic effect.Conclusion: Exosomes derived from ADSCs that have been osteogenic have a promoting effect on the osteogenesis of ADSCs that attached to silk fibroin-chitosan 3D scaffold in vitro.