The role for butyrate and propionate in mediating HeLa-cells growth inhibition by human dental plaque fluid from adult periodontal disease

Abstract

To determine to what extent propionate and butyrate, secreted in vitro by various oral bacteria, account for the growth-inhibiting activity of plaque fluid in gingivitis in the absence or presence of chronic periodontitis, the fluid phase of plaque from each of these disease forms was obtained by homogenizing the bacteria with 0.13 M NaCl, centrifuging the mixture and Millipore-filter-sterilizing the plaque-fluid-extract supernatant. Toxins were detected and measured by growth inhibition of HeLa cells grown in minimal Eagle medium supplemented with calf serum and antibiotics. Samples of butyrate, propionate or plaque-fluid extract were added and inhibition of growth was determined relative to control cultures. The toxins in each extract were characterized by their stability to heat, freeze-drying and elution on gel filtration. Butyrate and propionate contents were measured with a Hewlett-Packard S880A gas chromatograph. The plaque-fluid extracts from each group had similar contents of butyrate, but most of the activity was due to heat-sensitive high mol. wt components. Butyrate and propionate accounted for only 5 to 15 per cent of the total activity. High mol. wt exotoxins, rather than butyrate or propionate, were the most prominent and freeze-drying-stable tissue-destructive agents in the plaque fluid from subjects with or without periodontitis.