| Objective: The Craniomaxillo-facial bone defect caused by various reasons is a common clinical disease in plastic surgery.This experiment is based on the three elements of bone tissue engineering models include seed cells(rat adipose-derived stem cells);bioactive factors(rADSCs derived Exosomes);scaffold materials(nano-hydroxylapatite-collagen/Hydroxyacetic Acid poly(lactic-co-glycolic acid)scaffold,and verify the superiority of Exosomes repairs bone defect in vitro by implanted the model into rat skull defects.Methods: 1.Extracted the rADSCs and cultured it for Cell Proliferation,and induce it osteogenic and adipogenesis.2.Extracting the rADSCs induced by Osteogenesis,observing the morphology and staining the process of rADSCs internalized by Exos.3.Making the n HAC/PLGA scaffolds and test it’s biocompatibility and mechanical properties.Grouping by exert different factors.Include rADSCs suspension with scaffolds group,rADSCs suspension combine Exos suspension loaded with scaffolds group,simple scaffolds group and blank control group.Then implanted them into the skull defect of rats model,investigate the osteogenic activity of each group by Micro-CT and HE staining at4,12 weeks after surgeryed.Results: 1.rADSCs performed proliferate and differentiate well in vitro.Large Calcium nodules could be observed by osteoinduction after alizarin red staining.The formation of Lipid droplets was observed by adipogenesis induction after oil red O staining.2.The exosomes derived from rADSCs induced by osteogenesis were successfully extracted by ultracentrifugation and fluorescent stained to prove that the exosomes could be internalized by rADSCs.3.Compared with the other groups,rADSCs suspension combine Exos suspension showed better osteogenic activity by Micro-CT observe and HE staining in Vivo.Conclusion: The Exos combined rADSCs showed good osteogenesis in rat skull defect,and it is expected to be a new method for bone defect repair in clinic. |
