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Human epithelial cell death caused by Actinobacillus actinomycetemcomitans infection

Department of Periodontology, School of Dentistry, Health Sciences University of Hokkaido, Hokkaido and *Department of Oral Science, National Institute of Infectious Diseases, Tokyo, Japan

Corresponding author: Professor Y. Kowashi (e-mail: ykowashi{at}hoku-iryo-u.ac.jp).

Received 7 July 1999; revised version received 20 Nov. 1999; accepted 22 Dec. 1999.

Abstract

The gingival sulcus is the shallow crevice around the tooth, and its epithelium is a gateway for initial bacterial infection in periodontal disease. Recent studies have shown that Actinobacillus actinomycetemcomitans invades an epithelial cell line, KB cells, in vitro. The aim of the present study was to clarify the changes in KB cells after A. actinomycetemcomitans infection. The cytotoxic effects of A. actinomycetemcomitans on KB cells were determined at 72, 96 and 120 h after infection by an MTT assay. Nuclear morphological changes were observed by staining with Hoechst 33258. Cytoplasmic histone-associated DNA fragmentation in the infected KB cells was determined by ELISA. A. actinomycetemcomitans was cytotoxic on KB cells, and condensation and degradation of the nuclei were observed. DNA fragmentation was increased after the infection. In addition, A. actinomycetemcomitans showed similar cytotoxic effects on human gingival epithelial cells. The present study demonstrated that A. actinomycetemcomitans induces apoptotic cell death of oral epithelial cells in an in-vitro culture system. This induced apoptosis might be involved in the initiation and progression of periodontitis.

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