牙龈卟啉单胞菌脂多糖对牙周韧带细胞损伤模型愈合的影响

Effects of Pg-LPS on periodontal ligament cells in a wound healing model

目的研究不同浓度牙龈卟啉单胞菌脂多糖(Pg-LPS)对牙周韧带细胞(PDLCs)损伤模型愈合的影响。方法采用组织块法体外培养人PDLCs,取生长良好的第4代PDLCs接种于12孔板,待细胞融合后制备直径约8mm的圆形损伤模型,加入含不同浓度Pg-LPS的培养液培养,结晶紫染色观察损伤区细胞愈合情况、并检测损伤后各组细胞的数量变化;ELISA法检测初期损伤愈合时PDLCs分泌IL-1β、IL-6、TNF-α水平的变化。结果低浓度(0.001～1μg/mL)Pg-LPS对损伤愈合时PDLCs的增殖无明显影响(P>0.05);而高浓度(10μg/mL)Pg-LPS刺激可显著抑制PDLCs的增殖(P<0.05),延缓损伤模型的愈合,还可明显增加IL-1β、IL-6、TNF-α的分泌水平(P<0.05)。结论 Pg-LPS对PDLCs增殖的抑制作用主要与其浓度有关;高浓度Pg-LPS可促进IL-1β、IL-6、TNF-α的分泌。

Objective To evaluate the influence of different concentrations of Pg-LPS on PDLCs using an in vitro model of wound healing.Methods PDLCs were isolated,cultured by tissue block culture.After 4 passages,PDLCs were cultured in 12-well plates,and an in vitro wound model was mechanically established by removing cells in an 8mm diameter area.Then PDLCs were incubated in the presence of different concentration of Pg-LPS（0,0.001,0.01,0.1,1 and 10 μg/mL）,and the cell density and wound area for each group were evaluated.Levels of IL-1β,IL-6 and TNF-α on 1 d,3 d and 6 d were detected by ELISA.Results The growth inhibitory effects of Pg-LPS were statistically significant in 10 μg/mL group（P0.05） with increased secretion of IL-1β,IL-6 and TNF-α（P0.05）.Conclusion The inhibitory effects on PDLCs proliferation are related to the concentration of Pg-LPS.High concentration of Pg-LPS could promote the secretion of IL-1β,IL-6,TNF-α by PDLCs.