Department of Diagnostic Sciences, University of Texas Dental Branch at Houston, Houston, TX, USA.
J Periodontol. 2011 Jun;82(6):909-16. doi: 10.1902/jop.2010.100501. Epub 2010 Dec 28.
Porphyromonas gingivalis is etiologically associated with chronic periodontitis. The major fimbriae of this periodontal pathogen mediate binding to host gingival epithelial cells and fibroblasts, a critical function in the initiation of periodontitis. However, the role of fimbriae in P. gingivalis-osteoblast interactions remains unknown. In the present study, the involvement of major fimbriae in the initial and long-term interactions between P. gingivalis and osteoblasts is investigated.
Primary mouse calvarial osteoblast cultures were established and inoculated with P. gingivalis ATCC 33277 or YPF1, a major fimbriae-deficient mutant of P. gingivalis. Confocal microscopy images were acquired to assess bacterial invasion. DNA content measurement, real-time polymerase chain reaction, and alizarin red S staining and calcium content analysis were used to study the impact of bacteria on the proliferation, differentiation, and mineralization of osteoblasts, respectively.
Compared to the parent strain, YPF1 was significantly reduced in invasion of osteoblasts after 3 hours interaction. However, extended culture of infected osteoblasts did not reveal significant differences in persistence between the two strains. Proliferation of osteoblasts was not affected by either strain, and differentiation and mineralization of osteoblasts were inhibited by both strains to comparable levels.
This study reveals that major fimbriae are involved in the initial invasion of osteoblasts by P. gingivalis, but are not essential for the subsequent inhibition of osteoblast differentiation and mineralization in long-term culture.
牙龈卟啉单胞菌与慢性牙周炎在病因上有关。这种牙周病原体的主要菌毛介导与宿主牙龈上皮细胞和成纤维细胞的结合,这是牙周炎发病的关键功能。然而,菌毛在牙龈卟啉单胞菌-成骨细胞相互作用中的作用尚不清楚。本研究探讨了主要菌毛在牙龈卟啉单胞菌与成骨细胞的初始和长期相互作用中的作用。
建立了原代小鼠颅盖骨成骨细胞培养物,并接种牙龈卟啉单胞菌 ATCC 33277 或牙龈卟啉单胞菌 YPF1,后者是一种主要菌毛缺陷的突变株。通过共聚焦显微镜图像评估细菌入侵。通过 DNA 含量测定、实时聚合酶链反应、茜素红 S 染色和钙含量分析,分别研究细菌对成骨细胞增殖、分化和矿化的影响。
与亲本菌株相比,YPF1 在与成骨细胞相互作用 3 小时后,入侵成骨细胞的能力明显降低。然而,感染成骨细胞的延长培养并未显示两种菌株之间在持久性方面有显著差异。两种菌株均不影响成骨细胞的增殖,且两种菌株均能抑制成骨细胞的分化和矿化,抑制程度相当。
本研究表明,主要菌毛参与牙龈卟啉单胞菌对成骨细胞的初始入侵,但在长期培养中对成骨细胞分化和矿化的后续抑制并不必需。