尼古丁通过调控Toll样受体4抑制牙周膜干细胞的成骨分化能力

ObjectiveTo explore the impact of nicotine on proliferation and osteogenic capability of periodontal ligament stem cells (PDLSCs), and the role of Toll-like receptor 4 (TLR4) in nicotine, suppressing the osteogenic capability of PDLSCs.MethodsPDLSCs were culturedin vitro, and the flow cytometer was used to identify the surface antigen markers of PDLSCs. WST-1 was used to detect the proliferation ability of PDLSCs, which were stimulated by different concentrations of nicotine. Alizarin red staining was used to observe the formation of mineralized nodules after PDLSCs stimulation with different con-centrations of nicotine. Real-time polymerase chain reaction (RT-PCR) and Western blot were used to detect the change in osteogenic potential of PDLSCs stimulated by nicotine, after TAK-242, and with the inhibitor of TLR4.ResultsPDLSCs expressed mesenchymal stem cell-associated mar-kers CD90 and CD105. When the concentration of nicotine was 10^-4mol·L^-1, the PDLSC proliferation could be suppressed after 3 d compared with the control group (P<0 .05). the amount of mineralized nodules reduced after osteogenic differentiation at 21 d by alizarin red staining. rt-pcr and western blot showed the expression levels of alkaline phosphatase (alp), and osteocalcin (ocn), and the runt-related transcription factor-2 (runx-2) were lower than in the control group when nicotine suppressed the pdlscs (P<0 .05). this effect was atte-nuated after tak-242 was added.ConclusionNicotine suppresses the proliferation and osteogenic capability of PDLSCs, which may be regulated by TLR4.