Experimental Study On The Effects Of Aspirin-induced Treg Expression On The Proliferation And Osteogenic Differentiation Of Canine PDLSCs

Objective: Objective: To explore the effects of Aspirin-induced regulatory T cell(Treg)expression on the proliferation and osteogenic differentiation of canine periodontal ligament stem cells(PDLSCs).Methods: Flow cytometry was used to detect the changes in the ratio of Treg cells before and after oral administration of Aspirin in Beagle dogs.PDLSCs were successfully isolated and cultured in vitro and co-cultured in Transwell chambers.The study is divided into three groups.Group A is the control group PDLSCs,group B is the PDLSCs/Treg-(before induction)co-culture group,and group C is the PDLSCs/Treg+(after induction)co-culture group.Each group shares a certain ratio.After 72 hours of culture,real-time quantitative polymerase chain reaction(RT-PCR)and enzyme-linked immunosorbent assay(ELISA)were used to detect the expression of bone formation-related transcription factors(BMP2,Run X2,OCN).CCK-8 was used to detect cell proliferation;the osteogenic induction fluid was cultured for 14 days,and the degree of osteogenic differentiation was observed by staining with Alizarin Red.Results: Aspirin can induce an increase in the ratio of Treg cells in Beagle dogs;CCK-8 showed that the cell proliferation rate of the three groups was not statistically significant(P>0.05).After the sixth day,the three groups all entered the plateau phase,and the C group was higher than A The proliferation of the two groups was slightly higher,but the difference was not significant(P>0.05);14 days after osteogenic induction,the results showed that Treg cells can promote the osteogenic differentiation of PDLSCs,and the staining rate of Alizarin Red staining was significantly increased;RT-PCR,ELISA test showed that the expression of bone formationrelated transcription factors(BMP2,Run X2,OCN)increased significantly after induction.Conclusion: After induction,Treg cells can affect the osteogenic differentiation of PDLSCs,and can significantly up-regulate the staining rate of Alizarin Red,and up-regulate the expression of osteogenic-related transcription factors.