Fluorescence image analysis of the association between Porphyromonas gingivalis and gingival epithelial cells
Fluorescence image analysis of the association between Porphyromonas gingivalis and gingival epithelial cells
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DOI：
10.1046/j.1462-5822.1999.00022.x
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发表时间：
1999-11-01
影响因子：
3.4
通讯作者：
Lamont, RJ
中科院分区：
文献类型：
--
作者：
Belton, CM;Izutsu, KT;Lamont, RJ
我们已经开发了一种荧光成像技术，使用DNA结合染料可视化，随着时间的推移，牙龈卟啉单胞菌和人牙龈上皮细胞在体外的物理相互作用。该结果扩展了先前基于间接测量的牙龈卟啉单胞菌侵入牙龈上皮细胞的观察结果。牙龈卟啉单胞菌的细胞内定位通过在z平面中的图像的光学切片来建立。动力学分析表明，牙龈卟啉单胞菌的上皮细胞的入侵是一个快速和有效的过程，12分钟后完成。感染的单层成像显示，超过90%的牙龈上皮细胞的人口含有细菌。此外，只有活菌能够入侵，细胞内的细菌聚集在上皮细胞的核周区域。牙龈卟啉单胞菌在上皮细胞内停留24 h后，可诱导肌动蛋白细胞骨架沿着重排，并改变上皮细胞的大小和形状。这些发现提供了直接的证据，表明牙龈卟啉单胞菌进入牙龈上皮细胞的速率高且快速，并且内化的细菌最初定位于上皮细胞的特定区域。
We have developed a fluorescence imaging technique using a DNA-binding dye to visualize, over time, the physical interactions between Porphyromonas gingivalis and human gingival epithelial cells in vitro. The results extend previous observations of P. gingivalis invasion of gingival epithelial cells based on indirect measurements. An intracellular location for P. gingivalis was established by optical sectioning of images in the z-plane. Kinetic analysis showed that P. gingivalis invasion of epithelial cells is a rapid and efficient process, reaching completion after 12 min. Imaging of infected monolayers revealed that over 90% of a population of gingival epithelial cells contained bacteria. Furthermore, only vital bacteria were capable of invasion, and intracellular bacteria congregated in the perinuclear region of the epithelial cells. P. gingivalis remained inside the epithelial cells over a 24 h period and induced rearrangement of the actin cytoskeleton along with alteration of the size and shape of the epithelial cells. These findings provide direct evidence that entry rates of P. gingivalis into gingival epithelial cells are high and rapid, and that internalized bacteria initially localize in a specific region of the epithelial cells.