Effects of cecropin-XJ on growth and adherence of oral cariogenic bacteria in vitro.

Cecropin-XJ belongs to cecropin-B, which is the most potent antibacterial peptide found naturally. The aim of this study was to investigate the effects of cecropin-XJ on growth and adherence of oral cariogenic bacteria. Four oral cariogenic bacteria (Streptococcus mutans, Lactobacillus acidophilus, Actinomyces viscosus and Actinomyces naeslundii) were chosen for this experiment. The minimal inhibitory concentrations (MICs) and reductive percent of bacterial growth were used to assay the antibacterial activity of cecropin-XJ. Mammalian cytotoxicity of cecropin-XJ was tested with human periodontal membrane fibroblasts by tetrazolium (MTT) colorimetric assay. The bacterial morphological changes induced by cecropin-XJ were examined on scanning electron microscope (SEM). The influence of cecropin-XJ on bacterial adhesion to saliva-coated hydroxyapatite (S-HA) was measured by scintillation counting. The MICs of cecropin-XJ for inhibition of the growth of four bacteria ranged from 4.0 to 42.8 micromol/L with the highest susceptible to A. naeslundii and the lowest susceptible to L. acidophilus. At pH 6.8, 5.5 and 8.2, 1/2 MIC of cecropin-XJ reduced the number of viable bacteria by 40.9%, 67.8% and 32.8% for S. mutans and by 28.1%, 57.2% and 37.9% for L. acidophilus. The activities against S. mutans and L. acidophilus increased at pH 5.5 compared with pH 6.8 (P < 0.01, respectively). In present of 50% saliva, 1/2 MIC of the peptide decreased the direct count of viable cells by 29.2% and 14.4% for S. mutans and L. acidophilus, respectively (P < 0.01 and P > 0.05, respectively), whereas almost no reduction counts were detected in the presence of 20% serum for both bacteria (P > 0.05, respectively). Mammalian cytotoxicity of cecropin-XJ from 1.0 to 100 micromol/L exhibited no cytotoxicity against human periodontal membrane fibroblasts (P > 0.05). Bacterial morphological changes induced by MIC of cecropin-XJ examined on SEM showed cell surface disruption. Furthermore, the ability of A. naeslundii adhesion to S-HA decreased significantly with MIC of cecropin-XJ for 10 and 20 minutes (P = 0.001 and 0.000, respectively), and S. mutans, A. viscosus to S-HA decreased significantly with MIC of cecropin-XJ for 20 minutes (P = 0.000, respectively). Cecropin-XJ exhibited bactericidal action against cariogenic pathogens, and the antibacterial activity enhanced in the acid environment. The results also demonstrate that cecropin-XJ prevents S. mutans and actinomyces adsorption to S-HA. These findings suggest that Cecropin-XJ may have potential to prevent caries.