Department of Research Center, Faculty of Dentistry, Hacettepe University, Ankara, Turkey.
Republic of Turkey Ministry of Health, Oral and Dental Health Center, Ankara, Turkey.
Cytokine. 2021 Feb;138:155380. doi: 10.1016/j.cyto.2020.155380. Epub 2020 Nov 29.
Lipopolysaccharide is a potent virulence factor of Porphyromonas gingivalis and has been implicated predominant pathogen in the development and progression of periodontal diseases. The aim of this study was to determine the effect of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on cementoblasts. Cementoblast (OCCM-30) were evaluated proliferation using real-time cell analyzer. In addition, total RNA was isolated at 8, 16, 24 and 72 h from 1000 ng/mL Pg-LPS treated OCCM-30 cells and mRNA expressions of pro/anti-inflammatory cytokine mediators, extracellular matrix enzymes and their tissue inhibitors and of oxidative stress enzymes were studied by real-time polymerase chain reaction. Proliferation analysis indicated that Pg-LPS slightly decreased proliferation of OCCM-30. Pg-LPS had a time-dependent impact on the expression of cytokines and enzymes. There was statistically significant up-regulation of IL-1β and IL-10 in response to Pg-LPS at 8, 16, 24, 72 h but IL-6 expression was reduced compared to control at 8 h. While IL-8 and IL-17 expressions were determined higher than control group at 16 and 24 h, their expressions were decreased compared to control groups at 72 h (p < 0.01). While MMP-1, MMP-2, MMP-3, TIMP-1, TIMP-2 expressions increased, MMP-9 expression reduced at time-points. Also, a time-dependent up-regulation in mRNA levels for oxidative stress enzymes was detected. These results indicated that up-regulation in the transcripts of inflammation-associated cytokines and degradation enzymes were noted in the cementoblasts exposed to Pg-LPS. Cementoblasts infected with the virulence factors of periodontopathogens might also involve to the induction of inflammation and degradation of the periodontal tissues.
脂多糖是牙龈卟啉单胞菌的一种强效毒力因子,并且被认为是牙周病发展和进展的主要病原体。本研究旨在确定牙龈卟啉单胞菌脂多糖(Pg-LPS)对成牙骨质细胞的影响。使用实时细胞分析仪评估成牙骨质细胞(OCCM-30)的增殖。此外,从用 1000ng/mL Pg-LPS 处理的 OCCM-30 细胞中分离总 RNA,并在 8、16、24 和 72 小时时通过实时聚合酶链反应研究促炎/抗炎细胞因子介质、细胞外基质酶及其组织抑制剂和氧化应激酶的 mRNA 表达。增殖分析表明,Pg-LPS 轻微降低了 OCCM-30 的增殖。Pg-LPS 对细胞因子和酶的表达具有时间依赖性影响。在 8、16、24 和 72 小时时,IL-1β和 IL-10 的表达呈统计学显著上调,但与对照组相比,8 小时时 IL-6 的表达降低。虽然在 16 和 24 小时时 IL-8 和 IL-17 的表达高于对照组,但与对照组相比,72 小时时其表达降低(p<0.01)。在 MMP-1、MMP-2、MMP-3、TIMP-1 和 TIMP-2 的表达增加的同时,MMP-9 的表达在时间点降低。此外,还检测到氧化应激酶的 mRNA 水平呈时间依赖性上调。这些结果表明,暴露于 Pg-LPS 的成牙骨质细胞中炎症相关细胞因子和降解酶的转录物上调。感染牙周病病原体毒力因子的成牙骨质细胞也可能参与诱导炎症和牙周组织的降解。