Effects Of MT01 On The MRNA Expression Of Specific Osteoblast-related Factors In Osteoblast MG63 Infected By Porphyromonas Gingivalis

Periodontitis is considered a result of local immune imbalance leading to periodontal tissue destructive disease,the relationship between the progress and severity of the disease and the host’s own immune inflammatory response is closely.Porphyromonas gingivalis destroy the periodontal tissueis by evading the host’s innate immune response to bacteria and secreting virulence factor, which is widely recognized as periodontal pathogens.In vitro studies have shown that Pg can invade the osteoblasts which maturation and mineralization is strained,reduce the formation of the alveolar bone, cause alveolar bone absorption.Therefore, it has clinical significance to resist the influence of Pg on osteoblasts, for treatment of periodontitis.MT01 is a kind of immunosuppressive oligodeoxynucleotide designed on the basis of human mitochondrial DNA,which can inhibit the activation of the body’s innate immune response caused by Toll-like receptor 9. We confirmed in our previous studies, MT01 can reduce the absorption of the alveolar bone of experimental rats caused by periodontitis;In vitro studies have shown that MT01 can upregulate the gene expression of osteoblast-related factors Runx2,SP7 in humanosteoblast-like cells MG63,and promote the differentiation of osteoblasts.At present,it has not yet been confirmed that whether MT01 as a kind of immunosuppressant has the effects on the differentiation of osteoblasts in infected status. This study by detecting MT01 intervened,the m RNA expression of specific osteoblast-related factors Runx2,SP7 in osteoblast MG63 infected by porphyromonas gingivalis,discuss whether MT01 has the effects on the differentiation of osteoblasts in infected status.The results showed that the m RNA levles of osteoblastrelated factors Runx2,SP7 in osteoblast MG63 were markedly upregulated by MT01 after infected by Pg or not,indicating that MT01 could still promot the differentiation of osteoblasts whether in infected status.Methods:The human osteoblast-like cells MG63 in a stable state were seeded to 6 well plates.First,specific sequence oligodeoxynucleotide MT01 at a concentration of 1 μg·m L-1 and the same volume of PBS were added separately,and the cells were incubated for 3 hours.Then bacterium suspension of Pg as multiplicity of infection MOI = 100:1 was added.The same volume of PBS was added in control group.There were four experimental groups:MT01+Pg,MT01,Pg and blank control group.Real-time quantitative polymerase chain reaction was used to detect them RNA expression of osteoblast-related factors Runx2,SP7 at 2,4,6,8,12,24 hours.Result:The m RNA levles of osteoblast-related factors in MG63 were markedly upregulated by MT01 after infected by Pg or not.However,there exsited differences in the ability of MT01 to upregulate the expression of Runx2, SP7 at different time-points.Conclusion:MT01 could promot the differentiation of osteoblasts in infected status.