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Detectable Dioxins in Human Saliva and Their Effects on Gingival Epithelial Cells

¹ Department of Oral Microbiology, Asahi University School of Dentistry, 1851-1 Hozumi, Mizuho, Gifu 501-0296, Japan; and
² Shimadzu Techno-Research, Inc., 1 Shimoai-cho, Nishinokyo, Nakagyo-ku, Kyoto 604-8436, Japan;

View larger version (48K): [in this window] [in a new window]   Figure 1. Measurement of dioxins in saliva specimens. Coplanar PCBs (A,B) and those with WHO-defined TEFs (C,D) in saliva (A,C) and blood (B,D) specimens were measured with the use of an HRGC/HRMS with a DB-5MS column. Twenty-two markers of PCBs were added to these specimens from eight subjects as internal standards. PCDDs with WHO-defined TEFs found in saliva (E) and blood (F) specimens were measured by means of an HRGC/HRMS with SP2331 and DB-17HT columns. Nineteen PCDD markers were added to these specimens as internal standards. The MS was operated in selected ion monitoring mode at a resolution of > 10,000. ND: not detected.

View larger version (45K): [in this window] [in a new window]   Figure 2. AhR mRNA expression in HGEC and human PBMC. AhR mRNA expression was analyzed by RT-PCR as described in MATERIALS & METHODS. The β-actin gene was assayed as a positive control, and PCR products of non-RT samples were examined as a negative control. Following PCR, a 10-µL quantity of the total amplified product underwent electrophoresis on ethidium-bromide-stained 1% agarose gels, and was visualized under ultraviolet fluorescence. Data presented represent three independent experiments.

View larger version (13K): [in this window] [in a new window]   Figure 3. IL-8-producing activity and cytotoxicity of PCBs and OCDD. HGEC were stimulated with 10-8 M of PCBs and 10-12 M of OCDD at 37°C for 24 hrs. After incubation, the supernatants were collected, and IL-8 production was determined by ELISA. Experiments were performed at least 3 times, with representative results presented. Each assay was done in triplicate wells, and the data are expressed as the mean ± SEM of results. Significant differences were seen between the groups with and without the test specimens (*P < 0.05). Cytotoxicity of the test specimens was also measured by LDH released into the culture supernatants. Percent cytotoxicity was calculated as detailed in MATERIALS & METHODS.

View larger version (20K): [in this window] [in a new window]   Figure 4. PCB 118 and OCDD augmented IL-8-producing activities stimulated with P. gingivalis fimbriae. HGEC were incubated with PCB 118 (A), OCDD (B), or medium alone, together with the indicated doses of P. gingivalis fimbriae (Pg fim) at 37°C for 24 hrs. After incubation, the supernatants were collected, and IL-8 production was determined by ELISA. Experiments were performed at least 3 times, with representative results presented. Each assay was done in triplicate wells, and the data are expressed as the mean ± SEM of results. Significant differences were seen between the groups with and without PCB 118 or OCDD (*P < 0.05).

Journal of Dental Research, Vol. 82, No. 10, 849-853 (2003)
DOI: 10.1177/154405910308201017


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