pH gradients induced by urea metabolism in 'artificial mouth' microcosm plaques

Evidence was sought for urea-induced pH gradients in dental plaque microcosm biofilms cultured from the mixed salivary bacteria in a multi plaque 'artificial mouth'. Application of 500 mmol/l urea for short periods (6 min) to 5-8 mm maximum-thickness plaques induced intraplaque pH gradients of up to 0.7 pH units with the surface alkaline relative to the inner plaque. These pH gradients persisted for more than 5 h in the absence of a flow of fluid. With 30-min urea applications and a flow of a basal medium containing mucin (BMM, pH 7.0), the pH of the inner (deeper) plaque regions also increased. Although the pH gradient initially formed was alkaline at the plaque surface, the BMM flow lowered the surface pH to neutrality whilst the inner layers were still alkaline, thereby reversing the pH gradient. In thick microcosm dental plaques, urea-induced pH gradients can therefore form and last many hours. They probably result from the significant time taken for urea to penetrate to the inner layers of plaque, its rapid metabolism by the outer plaque layers, and a rate-limiting clearance of ammonia. Even a slow BMM flow over the plaque greatly increased the rate of return to the resting pH, causing the gradients to change polarity.