环孢素A联合脂多糖对牙周膜成纤维细胞生物学活性的影响

Effects of Cyclosporine A and Porphyromonas gingivalis-lipopolysaccharide on Biological Activity of in vitro Cultured Human Periodontal Ligament Fibroblasts

目的探讨环孢素A(CsA)联合牙龈卟啉单胞菌脂多糖(Pg-LPS)对人牙周膜成纤维细胞(PDLFs)生物学活性的影响。方法将CsA(100 ng/mL)、LPS(100 ng/mL)分别单独及联合作用于人PDLFs,采用考马斯亮蓝染色、ELISA、Von kossa染色等方法检测其对PDLFs总蛋白量、碱性磷酸酶(ALP)、骨形成蛋白2(BMP-2)及矿化结节形成的影响。结果 CsA单独及与LPS联合应用可促进细胞总蛋白的合成,增加细胞ALP活性及BMP-2的表达,并促进细胞矿化结节的早期形成;LPS单独应用可增加细胞的ALP活性,但对细胞的总蛋白合成、BMP-2的表达及矿化结节的早期形成无明显影响。结论一定浓度的CsA促PDLFs合成、分化作用明显,CsA联合LPS在促进细胞分化方面具有一定的协同作用。

Objective To evaluate the in vitro effects of cyclosporine A （CsA） and Porphyromonas gingivalis-lipopolysaccharide（Pg-LPS） on the biological activity of human periodontal ligament fibroblasts （hPDLFs）. Methods hPDLFs were incubated with CsA 100 ng/mL, Pg- LPS 100 ng/mL,CsA 100 ng/mL＋Pg-LPS 100 ng/mL. Then the total protein was extracted and the concentration was assessed with Coomassie brilliant blue. Several features such as expression of bone morphogenetic protein-2（BMP-2） and alkaline phosphatase（ALP） were detected by enzyme-linked immunosorbent assay （ELISA）. The formation of mineralized islets was assessed by Von kossa staining. Results CsA alone or plus LPS could significantly enhanced the total protein concentration, ALP, BMP-2 and mineralized islets of hPDLFs; LPS alone could significantly promote cells ALP expression,whereas has no significant effect on protein concentration, BMP-2 and mineralized islets of hPDLFs. Conclusion At certain concentrations, CsA could significantly enhance hPDLFs biological activity, synergistic effect was observed on enhancing cell differentiation when combined with LPS.