Effect of smokers' sera on Porphyromonas gingivalis internalizing KB cells and the expression of matrix metalloproteinase-1,-9 and tissue inhibitor of metalloproteinase-1
10.3760/cma.j.issn.1002-0098.2014.01.005
- VernacularTitle:吸烟个体血清对牙龈卟啉单胞菌内化KB细胞及KB细胞产生基质金属蛋白酶1、9和金属蛋白酶组织抑制剂1的影响
- Author:
Hongyan WANG
1
;
Lisi TAN
;
Junchao LIU
;
Qian LI
;
Yaping PAN
;
Ming ZHONG
Author Information
1. 中国医科大学口腔医学院牙周科·辽宁省口腔医学研究所
- Keywords:
Smoking;
Porphyromonas gingivalis;
KB cells;
Matrix metalloproteinases
- From:
Chinese Journal of Stomatology
2014;49(1):15-20
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of serum from smoking individuals or non-smoking individuals with periodontitis on Porphyromonas gingivalis(Pg) internalizing KB cells,and the expression of matrix metalloproteinase(MMP)-1,MMP-9,tissue inhibitor of metalloproteinase-1 (TIMP-1) in the culture supernatant of KB cells.Methods The venous blood of 20 periodontitis patients'(10 smoking and 10 non-smoking) was extracted under the informed consent and centrifuged for serum.The smoking-individual serum(Y group) and non-smoking-individual(N group)serum were added to the model of Pg internalizing KB cells for 12 hours,plated on brain-heart infusion(BHI) and incubated anaerobically at 37 ℃ for 5 days.The colony forming units(CFU) of cell-invasive bacteria were estimated by colony counting.M MP-1,MMP-9 and TIMP-1 protein levels in culture supernatant were determined by enzyme-linked immunosorbent assay (ELISA) in the two groups following co-culture of Pg with KB cells for 12 hours.Results The CFU were (11.2 ± 1.1) × 104,(12.6 ± 1.2) × 104,(44.7 ± 1.3) × 104 CFU/ml when adding 200,400,800 μl Y-group serum to the model of Pg co-culture with KB cells and when the serum was extracted from N group,the CFU were (33.6 ± 1.4) × 104,(38.9 ± 1.1) × 104,(11.2 ± 1.2) × 104 CFU/ml respectively.When 200,400,800 μl Y group-serum was added to co-culture fluid of Pg internalizing KB cells,the concentrations of MMP-1 secreted from KB cells were (107.2 ± 21.5),(165.9 ± 20.2),(434.4 ± 48.0) μg/L respectively,the concentrations of M MP-9 were (3.99 ±0.29),(4.21 ± 0.61),(5.62 ± 0.47) μg/L respectively,the concentrations of TIMP-1 were (401.3 ± 12.7),(418.3 ± 28.5),(637.3 ± 37.3) μg/L.When the serum (200,400,800 μl) extracted from N group,the concentration of MMP-1 and MMP-9 secreted by KB cell were (77.6±10.8),(84.7 ± 10.2) and (98.2 ±9.7) μg/L and (3.84 ±0.52),(4.02 ±0.68),(4.25 ±0.37) μg/L,respectively.The concentration of TIMP-1 were (67.3 ±26.9),(89.4 ± 22.7) and (78.2 ± 16.5) μg/L secreted by KB cells in the course of Pg internalized KB cell.With the increasing of Y group-serum,the more MMP-1,MMP-9 and TIMP-1 were secreted by KB cells (P ＜ 0.05).When 800 μl Y group-serum was added compared with N group-serum to the Pg co-culture with KB model,the more MMP-1,MMP-9 and TIMP-1 were secreted by KB cells (P＜0.05),when 400 μl Y group-serum was added compared with N group-serum to the Pg co-culture with KB model,the more MMP-1 and TIMP-1 were secreted by KB cells (P ＜ 0.05).Conclusions The smoking-serum might enhance Pg internalizing KB cells and enhance the expression of MMP-1,MMP-9 and TIMP-1 secreted from KB cells.The local microenvironment of smoking individual may contribute to the recurrence and progression of chronic periodontitis.
