TREPONEMA-DENTICOLA INDUCES ACTIN REARRANGEMENT AND DETACHMENT OF HUMAN GINGIVAL FIBROBLASTS
TREPONEMA-DENTICOLA INDUCES ACTIN REARRANGEMENT AND DETACHMENT OF HUMAN GINGIVAL FIBROBLASTS
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DOI：
10.1128/iai.60.8.3360-3368.1992
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发表时间：
1992-08-01
影响因子：
3.1
通讯作者：
ELLEN, RP
中科院分区：
文献类型：
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作者：
BAEHNI, PC;SONG, M;ELLEN, RP
螺旋体与破坏性牙周疾病有关，并且一种可培养的口腔物种齿垢密螺旋体结合哺乳动物细胞并扰乱新陈代谢。探讨T.用T.无血清培养基中的齿垢菌菌株ATCC 35405、e和e '。HGF使伪足收缩，变圆，并最终从基底层脱离。扫描电子显微镜显示螺旋体与HGF表面密切接触;偶尔，细菌部分淹没在HGF膜的褶皱之间。随着HGF的收缩，细胞表面形成大量的微绒毛和气泡。通过共聚焦显微镜，螺旋体检测接触HGF表面，但从来没有发现之间的基质和细胞的成纤维细胞的腹面。形态学改变与肌动蛋白组装相关，并在此之前，通过显微镜荧光测定法测量：在30分钟内，肌动蛋白荧光比对照组增加了263%。成纤维细胞从基质中分离与孵育时间有关，并依赖于T的浓度。齿垢观察到所有测试菌株的分离，并且也依赖于T.齿垢：紫外线、热和甲硝唑治疗显著降低了HGF脱离反应。分离也显着减少蛋白酶抑制剂苯甲基磺酰氟。肝细胞生长因子的活力没有显着影响共孵育与螺旋体，测定乳酸脱氢酶释放。因此，T.齿垢诱导快速的细胞骨架重塑，随后是细胞脱离，这可能是由细菌相关的蛋白酶刺激的，但不太可能直接由细胞-基质粘附接触点的蛋白水解降解介导。
Spirochetes are associated with destructive periodontal diseases, and one cultivatable oral species, Treponema denticola, binds to mammalian cells and perturbs metabolism. To evaluate the cytoskeletal responses and attachment functions of human gingival fibroblasts (HGF) exposed to T. denticola, monolayers of HGF were incubated with T. denticola strains ATCC 35405, e, and e' in serum-free medium. HGF retracted pseudopods, rounded up, and ultimately detached from the substratum. Scanning electron microscopy showed spirochetes in close contact with HGF surfaces; occasionally, bacteria were partially submerged between folds in the HGF membrane. Blebbing and numerous microvilli formed on the cell surface as the HGF retracted. By confocal microscopy, spirochetes were detected in contact with the HGF surface but were never found on the ventral surface of fibroblasts between the substratum and cell. Morphological alterations were associated with and preceded by actin assembly, as measured by microscopic fluorimetry: there was a 263% increase in actin fluorescence over controls within 30 min. Detachment of fibroblasts from the substratum was related to incubation time and was dependent on the concentration of T. denticola. Detachment was observed for all strains tested and was also dependent on the viability of T. denticola: UV light, heat, and metronidazole treatment markedly reduced the HGF detachment response. Detachment was also significantly reduced by the protease inhibitor phenylmethylsulfonyl fluoride. HGF viability was not significantly affected by coincubation with spirochetes, as measured by lactate dehydrogenase release. Thus, T. denticola induces rapid cytoskeletal remodelling followed by cell detachment, which might be stimulated by a bacterially associated protease but is not likely directly mediated by proteolytic degradation at the cell-substratum adhesive contact points.