Recommendation for a non-animal alternative to rat caries testing.

PURPOSE: As a requirement of the Food & Drug Administration's final monograph on "Anticaries drug products for over-the-counter human use", the toothpaste industry has been conducting animal caries tests on every fluoride-containing toothpaste introduced into the U.S. market since 1996. The practice of testing in animals, although required by law, is in stark conflict with the corporate policy of many U.S. and global toothpaste manufacturers, in which, if possible, alternatives to animal testing are utilized. A provision does exist within the regulation which allows the use of an alternative method to demonstrate efficacy. However, to take advantage of this provision, a petition must be submitted to the FDA and in this petition data demonstrating the alternative provides results of "equivalent accuracy" must be included. After many years of research, model development and model comparisons, we have identified one particular laboratory model that demonstrated excellent correlation with the currently accepted animal caries models. This model, known as the Featherstone pH cycling model, is discussed in this paper. METHODS: The Featherstone pH cycling model has been shown to produce results of equivalent accuracy to the animal caries model by: (1) demonstrating a clinically relevant fluoride dose response similar to that shown in the animal caries model (including 1100 ppm F, 250 ppm F and placebo); (2) demonstrating similar results to the animal caries model for clinically proven dentifrice formulations relative to positive and negative controls; (3) demonstrating discriminating ability in strong agreement with the animal caries model for differentiating between a dentifrice formulation with attenuated fluoride activity and a USP standard; and (4) providing a clinically relevant representation of the caries process, as demonstrated by orthodontic banding studies. In addition, the model sufficiently addresses both salivary and abrasive/anticalculus agent interference concerns. RESULTS: For more than 50 years, fluoride has been the first defense in the fight against dental caries. The clinical effectiveness of fluoride is well accepted and documented extensively in the literature. The mechanism through which fluoride provides its benefit is very straightforward and well understood. The proposed laboratory model effectively simulates the effect of the caries process and has been shown to demonstrate equivalent accuracy to animal caries. This indicates that there are strong scientific grounds for the use of this model as an alternative to the animal caries test. Based on the strength of the data and the correlations noted between the two models, we recommend that the scientific community and the toothpaste industry broadly accept the Featherstone laboratory pH cycling model as an appropriate alternative to animal testing, particularly for ionic fluoride based dentifrices.

The Featherstone laboratory pH cycling model: a prospective, multi-site validation exercise.

PURPOSE: To demonstrate the robustness of the Featherstone pH cycling model when tested in three independent laboratories and to evaluate the use of "non-inferiority" testing at those laboratories. METHODS: The fundamental principles for the Featherstone laboratory pH cycling model to be an appropriate alternative to animal testing is that it must demonstrate equivalent accuracy to the "Gold Standard" (rat caries model) by: (1) providing a meaningful representation of the caries process; (2) demonstrating a proportionate response to fluoride dose (or concentration); (3) being able to show that clinically proven formulations perform similarly relative to the controls; and (4) differentiating products that have attenuated fluoride activity. RESULTS: This cross-validation study confirmed the ability of the three independent laboratories to discriminate between various concentrations of fluoride-containing dentifrice formulations, demonstrated that clinically proven formulas perform as expected and identified an attenuated fluoride formulation (NaF/CaCO3 dentifrice - 1100 ppm NaF) as inferior compared to the 1100 ppm F (NaF/silica) positive control.

Activity of amine oxide against biofilms of Streptococcus mutans: a potential biocide for oral care formulations.

AIMS: To assess the potential bactericidal activity of amine oxide (C10-C16-alkyldimethyl N-oxides) against Streptococcus mutans grown as planktonic suspension and as biofilm on hydroxyapatite discs, and its ability to control acidification of the media. METHODS: Amine oxide bacteriostasis was investigated using the Bioscreen C Microbiological Growth Analyser, while a standard suspension test was used to determine its bactericidal efficacy. In addition, the lethal activity of amine oxide was studied against sedimentation biofilms of S. mutans on hydroxyapatite (HA) discs and resuspended biofilms. Several parameters were considered such as the surfactant concentration, pH, the starting inoculum and the maturity of the biofilm. RESULTS: Amine oxide was bacteriostatic against planktonic S. mutans at a low concentration (0.006% v/v) and highly bactericidal against S. mutans in suspension or in a mature biofilm on hydroxyapatite, although the concentration required to achieve the latter effect was four times higher. The activity of amine oxide against biofilms depended upon its concentration and the age of the biofilm. In addition, amine oxide pre-treatment of the HA discs did not affect the growth of the biofilm. Finally, amine oxide did not prevent the acidification of the medium, although lower pHs had a potentiating effect on amine oxide activity. CONCLUSION: Amine oxide showed high potential for controlling early biofilms caused by periodontal bacteria. Further investigations should be carried out, particularly on the potential toxicity of amine oxide and its efficacy in complex formulations for oral care products.