AGEs Affect The Formation And Function Of Osteoclasts In Bone Regeneration Under High Glucose And High Fat Microenvironment

Objective : This research is a basic research on bone regeneration in orthopedic rehabilitation for T2 DM.It explores the effects of advanced glycation end products(AGEs)on osteoclasts(OC)in bone regeneration under high glucose and high fat microenvironment.The effect of generation and function,further analysis of the mechanism of AGEs in T2 DM bone regeneration.AGEs are expected to be used as a new therapeutic target in the rehabilitation treatment of delayed bone healing of T2 DM,and provide new ideas for clinical rehabilitation of T2 DM bone regeneration.Methods:(1)Using 50ng/m L nuclear factor kappa B ligand(RANKL)receptor activator and 20 ng/m L macrophage colony stimulating factor(M-CSF)to induce the differentiation of mouse RAW264.7 cells into OC in vitro,The induction experiment is divided into five groups: high glucose induction group;high glucose and high fat induction group;high glucose and high fat+AGEs(early stage intervention)induction group;high glucose and high fat +AGEs(advanced stage intervention)induction group;high glucose and high fat+AGEs(all stages intervention)induction group.(2)Use the anti-tartrate acid phosphatase(TRAP)staining method to identify and count the OC generated in the five groups after induction;(3)The cell proliferation detection kit(CCK8)was used to detect the proliferation activity of RAW264.7 cells in each group at different time points(day 1 to day 4);(4)The RAW264.7 cells were seeded on the bone resorption plate for induction culture,and the bone resorption capacity of each group of OC was evaluated by calculating the area of the bone resorption depression formed;(5)Western blot was used to detect the expression levels of TRAP,CTSK,MMP-9,and SDF-1 functional proteins in OC generated under different induction conditions.Results:(1)In the RAW264.7 cell induction experiment,AGEs can significantly increase the number of RAW264.7 cells differentiated into TRAP+ OC under high glucose and high fat microenvironment;(2)CCK-8 test results: the second,third,and fourth day,AGEs can increase the proliferation activity of RAW264.7 cells differentiated into OC under hyperglycemia and hyperlipidemia environment;(3)In the bone resorption experiment,the area of bone resorption pits in high glucose and high fat+AGEs(all stages intervention)induction group was significantly larger than that of the other groups,and the high glucose and high fat+AGEs(advanced stage intervention)induction group,high glucose and high fat+AGEs(early stage intervention)The area of bone resorption depression in the induction group increased significantly;(4)Western blot results showed that the expression levels of TRAP,MMP-9,CTSK,and SDF-1 in the induction group of high glucose and high fat+AGEs(all stages intervention)were significantly increased.Conclusion:(1)AGEs can promote the production and proliferation activity of OC under high glucose and high fat microenvironment;(2)AGEs can promote the bone resorption of OC under high glucose and high fat microenvironment,and the early stage intervention of AGEs is more effective than the advanced stage intervention;(3)AGEs can increase the expression of TRAP,MMP-9,CTSK and SDF-1 under high glucose and high fat microenvironment,thereby increasing the bone resorption capacity of OC.