Research Of EMMPRIN Expression And Relative Function In HPDLCs Under Mechanical Loading

Objective: 1.To identify the expression of EMMPRIN in HPDLCs under mechanical loading.2.To evaluate the correlation between EMMPRIN and VEGF in HPDLCs under mechanical loading.3.To explore the relationship between EMMPRIN and osteogenesis genes and the potential function of EMMPRIN in periodontal remodeling.Material and methods: 1.Primary periodontal ligament cells were obtained from healthy teeth.Immunofluorescent histochemical staining was performed to locate the expressions of EMMPRIN and VEGF in HPDLCs.HPDLCs were exposed to 6%,12% and 20% cyclic strain(strain treated,ST)and 0.6lbs compression force by the Flexcell System from 0.5h to 72 h.Changes of m RNA and protein expressions of VEGF in HPDLCs were observed after exogenous EMMPRIN protein was added(rh EMMPRIN treated,ET)into the HPDLCs culture medium.The changes of gene and protein expressions of EMMPRIN and VEGF in HPDLCs and protein in culture medium were observed by real-time PCR,Western Blotting,ELISA and CBA.2.Osteogenesis gene chip was used to screen the genes affected by rh EMMPRIN.Then,real-time PCR was performed to evaluate related genes expression treated with rh EMMPRIN under mechanical loading.Result: 1.6%,12% and 20% cyclic strain up-regulated EMMPRIN m RNA in similar trend and peaked at 48 h,while the EMMPRIN protein both in cells and culture mediumpeaked at 72 h when stimulated by 12% cyclic strain.The compression force didn't have specific regulation on EMMPRIN m RNA.2.12% cyclic strain up-regulated VEGF m RNA and peaked at 48 h.The expression of VEGF m RNA and protein both in HPDLCs and culture medium were up-regulated by rh EMMPRIN.Strain enhanced the upregulation of VEGF treated with rh EMMPRIN.3.rh EMMPRIN up-regulated ALP and PDGFA and down-regulated NOG and BGLAP expression,indicating that EMMPRIN may be involved in the differentiation of HPDLCs and potential bone remodeling.Conclusion: 1.Cyclic strain up-regulates the expression of EMMPRIN in HPDLCs.2.EMMPRIN participates in the vessel remodeling in periodontal tissue under stress through regulating VEGF and may involves in bone remodeling.