The effect of antibacterial monomer MDPB on the growth of organisms associated with root caries.

MDPB, 12-methacryloyloxydodecylpyridinium bromide, was tested for its ability to inhibit the growth of organisms associated with active root caries lesions and to modify the growth characteristics of these organisms at sub-MICs. MICs and MBCs of MDPB for independent isolates (n=5) of the following taxa: Streptococcus mutans, Streptococcus oralis, Streptococcus salivarius, Actinomyces naeslundii, Actinomyces israelii, Actinomyces gerensceriae, Actinomyces odontolyticus, Lactobacillus spp., and Candida albicans were determined, and the effects at sub-MIC on microbial growth kinetics were assessed. All isolates were sensitive to inhibition by MDPB. The median MICs and MBCs of MDPB for these organisms were in the range of 3.13 to 25.0 microg/ml and 6.25 to 50.0 microg/ml, respectively. As for the influence of pH, inhibition was sensitive to acidic pH. Even at sub-MICs, the growth of all strains, measured as cell yield and doubling time, was significantly reduced. Based on the results of this study, MDPB exhibited the potential to inhibit the growth of microbiota associated with active root caries lesions.

The influence of cholesterol, progesterone, 4-androstenedione and testosterone on the growth of Treponema denticola ATCC 33520 in batch cultures.

Previously, we have shown that reference and freshly isolated Treponema denticola cultures are capable of metabolising cholesterol, progesterone, 4-androstenedione and testosterone by means of 5alpha-reductase, 3beta-and 17beta-hydroxysteroid dehydrogenase activity [Clark DT, Soory M. The metabolism of cholesterol and certain hormonal steroids by Treponema denticola. Steroids. 2006;71:352-63. ]. The aim of the work presented in this paper was to investigate the modulation of T. denticola growth in batch cultures by these steroids, using T. denticola ATCC 33520 as a model system. Growth curves were summarised using statistics based on optical density and protein yield. Cholesterol was found to stimulate growth at concentrations of 10 and 25microg/mL. Certain hormonal steroids inhibited the maximum achievable optical density at concentrations of 1 and 10microg/mL while the minimum concentration shown to inhibit protein yield was 0.001microg/mL of progesterone. The potential of the hormonal steroids to inhibit growth was in the order of progesterone, 4-androstenedione and testosterone.

The metabolism of cholesterol and certain hormonal steroids by Treponema denticola.

The aim was to investigate whether reference cultures and fresh isolates of Treponema denticola are able to 5alpha-reduce and further metabolise testosterone, 4-androstenedione, progesterone, corticosterone, cortisol or cholesterol. Two reference and five freshly isolated cultures of T. denticola were incubated with either radiolabeled or unlabeled steroid substrates; in the first case products were identified by thin layer chromatography and in the latter by gas chromatography-mass spectroscopy. All the substrates were 5alpha-reduced. Both reference cultures and fresh isolates of T. denticola presented 3beta- and 17beta-hydroxy steroid dehydrogenase activity. It was concluded that T. denticola was capable of steroid metabolism and hypotheses are discussed regarding the in vivo function of this metabolism including, T. denticola utilising host supplied steroids as growth factors and T. denticola steroid metabolism acting as a virulence factor.

Effect of the environment on genotypic diversity of Actinomyces naeslundii and Streptococcus oralis in the oral biofilm.

The genotypic diversity of Actinomyces naeslundii genospecies 2 (424 isolates) and Streptococcus oralis (446 isolates) strains isolated from two sound approximal sites in all subjects who were either caries active (seven subjects) or caries free (seven subjects) was investigated by using the repetitive extragenic palindromic PCR. The plaque from the caries-active subjects harbored significantly greater proportions of mutans streptococci and lactobacilli and a smaller proportion of A. naeslundii organisms than the plaque sampled from the caries-free subjects. These data confirmed that the sites of the two groups of subjects were subjected to different environmental stresses, probably determined by the prevailing or fluctuating acidic pH values. We tested the hypothesis that the microfloras of the sites subjected to greater stresses (the plaque samples from the caries-active subjects) would exhibit reduced genotypic diversity since the sites would be less favorable. We found that the diversity of A. naeslundii strains did not change (chi2 = 0.68; P = 0.41) although the proportional representation of A. naeslundii was significantly reduced (P < 0.05). Conversely, the diversity of the S. oralis strains increased (chi2 = 11.71; P = 0.0006) and the proportional representation of S. oralis did not change. We propose that under these environmental conditions the diversity and number of niches within the oral biofilm that could be exploited by S. oralis increased, resulting in the increased genotypic diversity of this species. Apparently, A. naeslundii was not able to exploit the new niches since the prevailing conditions within the niches may have been deleterious and not supportive of its proliferation. These results suggest that environmental stress may modify a biofilm such that the diversity of the niches is increased and that these niches may be successfully exploited by some, but not necessarily all, members of the microbial community.