Abstract
BACKGROUND: Porphyromonas gingivalis, a gram-negative obligate anaerobic bacterium, is a major pathogen involved in the onset and progression of periodontal disease, a chronic inflammatory disorder observed in approximately two-thirds of the Japanese population older than age 30 years. P gingivalis cells produce and secrete gingipain, a powerful proteolytic enzyme, on their surfaces and in external environments. OBJECTIVES: The effects of heat-killed Enterococcus faecalis (HkEf), a lactic acid bacterium, on the growth of P gingivalis were evaluated in vitro by measuring the viable cell count of P gingivalis and gingipain activity. METHODS: HkEf solution (1.63 or 163 mg/mL) was added to 1 mL P gingivalis culture to generate a final HkEf concentration of 0.64 or 64 mg/mL. The cultures were incubated anaerobically. The number of viable P gingivalis cells and gingipain activity were measured after incubation for 0, 12, 24, 48, and 72 hours. The number of viable P gingivalis cells was calculated by counting the number of colonies after culture. Gingipain activity was quantified by adding a chromogenic substrate to P gingivalis culture medium and measuring the absorbance of the reaction solution with a plate reader. Mean (SE) was calculated for viable cell counts and gingipain activity, and Wilcoxon rank-sum test was used to test for significant differences. RESULTS: The counts of viable P gingivalis cells in the control group increased as incubation time progressed for 12, 24, 48, and 72 hours; similar results were observed in the low-concentration HkEf group. In the high-concentration HkEf group, the increase in the viable cell count was significantly inhibited compared with that of the control group. Furthermore, gingipain activity in the low- and high-concentration HkEf groups was significantly inhibited over time compared with that of the control group. Although the pH of the culture solution tended to decrease in the high-concentration HkEf group, it was not considered to have affected the growth of P gingivalis. CONCLUSIONS: HkEf exhibits inhibitory effects on the growth of P gingivalis and gingipain activity.