Effects Of Lentivirus-mediated XBP1s On Proliferative And Osteogenic Capacity Of Periodontal Ligament Cells

Object: Periodontitis is a chronic human inflammatory disease that is widespread worldwide,and there is accumulating evidence that many endoplasmic reticulum stress（ERS）-related genes are upregulated in periodontitis.The purpose of this study was to explore the survival and proliferation of human periodontal ligament cells（hPDLCs）in ERS model,and to study the related effects of ERS on hPDLCs.The effects of the spliced form of X-box binding protein-1（XBP1s）on the proliferation and osteogenesis of hPDLCs were detected by constructing a plasmid overexpressing XBP1 s and infecting hPDLCs with lentivirusMethod: hPDLCs were cultured in vitro and three groups were set: blank group（Minimum Essential Medium Alpha,MEM-α）,control group（Dimethyl sulfoxide,DMSO）and experimental group（Tunicamycin,TM）.Using Western Blot,proteins associated with the ERS pathway,such as e IF2α,GRP78 and XBP1 s,were detected.Then the hPDLCs overexpressing XBP1 s mediated by and lentivirus was constructed and Western Blot was used to detect autophagy-related P62 and LC3,as well as apoptosis-related Bcl-2 and Caspase-3;in addition the interaction between XBP1 s and human bone morphogenetic protein 2（BMP-2）was detected using indirect immunofluorescence antibody test（IFAT）;Furthermore,the senescence,proliferation and m RNA expression of osteogenic genes were detected using β-galactosidase staining,CCK-8 and RT-q PCR,respectively.Result: The results showed an increase in proliferation of hPDLCs from 0 to 24 h when ERS was induced（P<0.05）.pLVX-XBP1s-PDLCs were successfully established,XBP1 s were able to significantly induce proliferation（P<0.05）;as autophagy-related proteins,P62 and LC3II/LC3 I ratio were significantly decreased and increased,respectively（P<0.05）;as apoptosis-related proteins,Bcl-2 and Caspase-3 were significantly increased and decreased,respectively（P<0.05）;the ratio of senescent cells was markedly decreased（P<0.05）;after infection with BMP-2lentiviral supernatant,m RNA expression of osteogenesis-related genes including Runx2,COL I,OC and OPN was continually up-regulated（P<0.05）;otherwise,the m RNA of Runx2,COL I and OC were highly expressed（P<0.05）,except for OPN（P>0.05）.Conclusion: XBP1 s can play a positive role in the proliferation of PDLCs by regulating autophagy and inhibiting apoptosis.Meanwhile,XBP1 s can significantly promote the osteogenesis of hPDLCs significantly promote the proliferation and osteogenesis capacity of hPDLCs.Further exploration of the mechanisms in this regard is needed for periodontal tissue regeneration,functionalization and clinical applications.