Preliminary assessment of safety and effectiveness in humans of ProBiora3, a probiotic mouthwash.

AIMS: To conduct a pilot human clinical trial to assess the safety and to test the ability of a probiotic mouthwash, ProBiora(3), to affect the levels of Streptococcus mutans and certain known periodontal pathogens in the mouth when administered twice daily over a period of 4 weeks. METHODS AND RESULTS: The mouthwash contained three specific strains of naturally occurring oral bacteria and was tested at two dose levels: 10(6) and 10(8) colony forming units each of Strep. oralis strain KJ3sm, Strep. uberis strain KJ2sm, and the spontaneous lactic acid-deficient variant of Strep. rattus, strain JH145. Substantial decreases in the levels of the marker bacteria were observed. No safety issues were noted with the twice daily application of this mouthwash. CONCLUSIONS: Despite the small number of subjects and the use of young, orally healthy adults, along with the inherent variability in the microbiological measurements, the probiotic mouthwash was able to substantially affect the levels of dental pathogens in saliva and periodontal pathogens in subgingival plaque. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this pilot human study suggest that the probiotic mouthwash product may be safe for daily use as an aid in maintaining both dental and periodontal health.

A spontaneous lactate dehydrogenase deficient mutant of Streptococcus rattus for use as a probiotic in the prevention of dental caries.

AIMS: To study the ability of daily applications of Streptococcus rattus strain JH145 to affect the numbers of an implanted Streptococcus mutans strain in a rat model. METHODS AND RESULTS: A spontaneous L(+)-lactate dehydrogenase (LDH)-deficient mutant of Streptococcus rattus, JH146, was isolated by screening on selective medium and compared with a previously isolated spontaneous LDH deficient strain, JH145. Both strains were shown to have single base pair deletion mutations in the structural gene (ldh) for LDH, and reversion frequencies were approximately the same. Animals treated once daily with >or=10(6) CFU (colony forming units) of JH145 showed a statistically significant decrease in the proportion of implanted S. mutans to total cultivable bacteria in oral swab samples. The rate of decrease in S. mutans levels was dose-dependent. No adverse effects were observed by in-life observation of treated animals, and histopathological, haematological and blood chemistry analyses were unremarkable. CONCLUSIONS: The results presented indicate that daily application of JH145, a naturally occurring LDH-deficient variant of S. rattus, can compete with S. mutans for its habitat on the tooth surface. SIGNIFICANCE AND IMPACT OF THE STUDY: S. rattus JH145 has potential as a probiotic for use in the prevention of dental caries.

Evaluation of a chairside gingival protease test for use in periodontal diagnosis.

Collagen degradation is a major factor in the destruction of diseased periodontal tissues. Evidence from several investigations indicates that this connective tissue breakdown process may be associated with elevated levels of proteolytic activity in gingival crevicular fluid. A rapid, chairside assay for neutral proteases in biological fluids was evaluated clinically for reproducibility and an indicator of periodontal disease status. This colorimetric test is based on the use of an insoluble, covalently linked dye-collagen substrate. Enzymatic activity is monitored by estimating the production of soluble dye-labeled collagen fragments resulting from the action of sample fluid proteases on the test substrate. Gingival fluid samples were collected on three successive days from 366 interdental sites in 34 volunteers, with and without periodontal disease. Results demonstrate that the assay is reproducible with 98.6% of sampled sites remaining stable throughout the 3-day study. Furthermore, the highest percentage of elevated neutral protease activity (NPA) values and the highest NPA values were found in patients clinically diagnosed as having advanced periodontal disease.

Modification by salivary pellicles of in vitro enamel remineralization.

The effectiveness of inorganic calcifying solutions to remineralize enamel with subsurface demineralization decreased with saliva pre-treatments conducive to the formation of enamel pellicles. Greater reductions in remineralization rates occurred with longer pellicle formation times. It is suggested that enamel pellicles may act to control surface deposition and favor subsurface precipitation.

Demineralization and remineralization of dental enamel.

Experimentation in vitro using organic acid buffers as demineralizing media shows that caries-like lesions can be obtained which are very similar in morphology and developmental stages to early lesions formed naturally under oral conditions. The use of these chemical systems and of mechanistic models advanced to explain the unique histological features of incipient caries have yielded a good understanding of the processes involved in caries formation. The study of natural and induced factors influencing the demineralization process has been greatly facilitated by the use of bacteriological systems in which demineralization is produced by direct colonization of cariogenic microorganisms on the surfaces of extracted teeth. Comparison of results obtained with these latter systems and with chemical systems has allowed us, for example, to elucidate the mechanism by which acquired salivary pellicles and fluoride topical solutions decrease the rate of enamel demineralization. The pellicle retards transport of matter across the enamel surface, whereas the fluoride topical solutions decrease the cariogenicity of the colonizing bacteria.

Effect of salivary pellicle formation time on in vitro attachment and demineralization by Streptococcus mutans.

Enamel subsurface demineralization induced by Streptococcus mutans was significantly reduced by seven-day saliva pre-treatments conducive to the formation of enamel pellicles. A two-hour saliva pre-treatment was ineffective. Results suggest that the protection provided by long-term pellicles may relate to changes in ionic transport rates rather than cell attachment.

Effect of fluoride topical solutions on enamel demineralization by lactate buffers and Streptococcus mutans in vitro.

Applications of APF and NaF solutions to extracted human teeth reduced the extent of enamel subsurface demineralization induced by colonization of S. mutans but were ineffective against demineralization by acid lactate buffers. It is concluded that the CaF2 formed on the tooth surface during the treatment is responsible for the protection observed.

In vitro enamel demineralization by Streptococcus mutans in the presence of salivary pellicles.

Salivary pellicles developed on extracted teeth favorably affected the degree and nature of enamel demineralization when the teeth were incubated in vitro with either of two pure strains of cariogenic Streptococcus mutans. The mechanism responsible for this protection may relate to the permselective properties of these salivary pellicles.

Effect of salivary pellicle on enamel subsurface demineralization in vitro.

Diffusion fluxes of two anionic species through hydroxyapatite membranes were found to be reduced up to 50% by the presence of adsorbed salivary pellicles developed on the membrane surfaces. By contrast, water fluxes were only marginally affected, indicating that salivary pellicles display ionic permselectivity. This property is used to explain a remarkable protection of the enamel observed when salivary pellicles were developed on extracted teeth before exposure to acid lactate buffers.