Effect of Treponema lecithinolyticum lipopolysaccharide on matrix metalloproteinase-9 expression / 대한치주과학회지

Bone resorption involves sequential stages of osteoclast precursor migration and differen-tiation of osteoclast precursors into multinucleated osteoclasts. Stromal cell derived factor (SDF)-1 is a chemotactic factor for osteoclast precursor migration. Matrix metalloproteinase (MMP)-9 is involved in migration of osteoclast precursors and activation of interleukin(IL)-1beta. Alveolar bone destruction is a characteristic feature of periodontal disease. Treponema lecithinolyticum is a oral spirochete isolated from the periodontal lesions. The effect of lipopolysaccharide(LPS) from T. lecithinolyticum on expression of SDF-1 and MMP-9 was examined in cocultures of bone marrow cells and osteblasts derived from mouse calvariae. T. lecithinolyticum LPS increased expression of MMP-9 in the coculture. Polymyxin B, an inhibitor of LPS, abolished the increase of MMP-9 mRNA expression by LPS. LPS did not increase the expression of SDF-1, IL-1beta and tumor necrosis factor(TNF)-alpha mRNA in cocultures. Prostaglandin E2(PGE2) up-regulated the expression of MMP-9 and NS398, an inhibitor of PGE2 synthesis, down-regulated the induction of MMP-9 expression by T. lecithinolyticm LPS. These results suggest that T. lecithinolytium LPS increases MMP-9 expression in bone cells via PGE2 and that the induction of MMP-9 expression by T. lecithinolyticum LPS is involved in alveolar bone destruction of periodontitis patients by the increase of osteoclast precursor migration and the activation of bone resorption-inducing cytokine.

Effect of Treponema lecithinolyticum lipopolysaccharide on matrix metalloproteinase-9 expression / 대한치주과학회지

Bone resorption involves sequential stages of osteoclast precursor migration and differen-tiation of osteoclast precursors into multinucleated osteoclasts. Stromal cell derived factor (SDF)-1 is a chemotactic factor for osteoclast precursor migration. Matrix metalloproteinase (MMP)-9 is involved in migration of osteoclast precursors and activation of interleukin(IL)-1beta. Alveolar bone destruction is a characteristic feature of periodontal disease. Treponema lecithinolyticum is a oral spirochete isolated from the periodontal lesions. The effect of lipopolysaccharide(LPS) from T. lecithinolyticum on expression of SDF-1 and MMP-9 was examined in cocultures of bone marrow cells and osteblasts derived from mouse calvariae. T. lecithinolyticum LPS increased expression of MMP-9 in the coculture. Polymyxin B, an inhibitor of LPS, abolished the increase of MMP-9 mRNA expression by LPS. LPS did not increase the expression of SDF-1, IL-1beta and tumor necrosis factor(TNF)-alpha mRNA in cocultures. Prostaglandin E2(PGE2) up-regulated the expression of MMP-9 and NS398, an inhibitor of PGE2 synthesis, down-regulated the induction of MMP-9 expression by T. lecithinolyticm LPS. These results suggest that T. lecithinolytium LPS increases MMP-9 expression in bone cells via PGE2 and that the induction of MMP-9 expression by T. lecithinolyticum LPS is involved in alveolar bone destruction of periodontitis patients by the increase of osteoclast precursor migration and the activation of bone resorption-inducing cytokine.

Functional Characterization of the Major Outer Membrane Protein ( MspTL ) of Treponema lecithinolyticum

Treponema lecithinolyticum is known to be associated with rapidly progressive periodontitis. The full-length gene encoding the major surface protein MspTL of this organism was cloned and expressed by using the expression vector pQE30. Recombinant Msp TL (rMsp TL) protein was purified and functionally characterized. Msp TL was one of several T. lecithinolyticum surface proteins that bound to cultured human gingival fibroblasts (HGFs). As well, it strongly induced IL-6 production by HGFs. These observations suggest that MspTL contributes to pathogenesis of periodontitis by adhesion to host cells and by induction of the proinflammatory cytokine IL-6.

The Effect of Sonicated Extracts of Treponema Denticola and Treponema Lecithinolyticum on the Cytokine Secretion and Matrix Metalloproteinase Activation of Gingival Fibroblast / 대한치주과학회지

This study was investigated to observe the effect of Treponema denticola cell sonicates(TDC) and Treponema lecithinolyticum cell sonicates(TLC) on cytokine secretion and matix metalloproteinase-2(MMP-2) activation of cultured human gingival fibroblast. Several experiments were performed including IL-1beta, IL-6 ELISA for the effect on the IL-1beta, IL-6 secretion of human gingival fibroblast. Also gelatinase zymography and gelatin dissolubility test for the activation of MMP-2 secreted by gingival fibroblast. The results were as follows. 1.The effect of TDC and TLC on IL-6 secretion of human gingival fibroblast showed statistically significant increase of IL-6 secretion in the TDC and TLC treated group compared to no treatment group(p<0.05) . 2.The amount of IL-1beta secretion was below the lower limit and there was no difference in the IL-1beta secretion of gingival fibroblast between TDC, TLC treated group and no treatment group. 3.The active form of pro MMP-2 with 72 kDa molecular weight was activated in both TDC and TLC treated group and clear band was appeared at 62kDa site on the zymography. 4.Gelatin dissolubility of MMP-2 secreted by gingival fibroblast was higher in TDC and TLC treated group compared to no treatment group(p<0.05). 5.In the TDC treated group, serine protease of T. denticola affect gelatin dissolubility. But in the TLC treated group gelatin was degraded by only MMP secreted by gingival fibroblast. Regarding to the above results, TDC and TLC have an effect on the IL-6 secretion increase of human gingival fibroblast and appears to activate pro MMP-2 which degrades collagen.

The Effect of Treponema Denticola and Treponema Lecithinolyticum on Periodontal Ligament Cells / 대한치주과학회지

This study was investigated to observe the effect of Treponema denticola(TDC) and Treponema lecithinolyticum(TLC) on cultured human periodontal ligament cells. Several experiments were performed including MTT test for the inhibition effect of cell proliferation, LDH test for the cytotoxicity , gelatin zymography for the gelatinase activation and observation of cell morphology change using the phase-contrast microscopy. The results were as follows. 1. The effect of concentration on cell proliferation with time showed an inhibitory effect at high concentration(150microgram/well) for TLC and at low concentration( 9.4microgram/well ) for TDC. 2. The effect of time on cell proliferation with concentration showed an inhibitory effect at 150microgram/well on 2-day incubation for TLC and at 9.4microgram/well on 2-day incubation for TDC. 3. The effect of heat-treated TDC and TLC on the inhibition of cell proliferation showed the difference in the heat-treated group compared to the non-heat treated group for TDC, whereas no difference was found for TLC. 4. The morphological changes which were observed from the phase-contrast microscopy showed the difference in the test group compared to the control group. The loss of spindle-like appearance, cell-to-cell detachment and inhibition of cell proliferation were observed. 5. There was no difference of the cytotoxicity effect between the test group and the control group in the LDH test. 6. The active form of progelatinase A with molecular weight 72kDa was activated in both TDC and TLC on the gelatin zymography. Regarding to the above results, TDC and TLC have an effect on periodontal ligament cells by playing an inhibitory role in cell proliferation and appears to activate progelatinase A which degrades type IV collagen.