Abstract
BackgroundStreptococcus mutans is a common cariogenic bacterium in the oral cavity involved in plaque formation. Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) has been introduced into tooth mousse to encourage remineralization of dental enamel. The aim of this research was to study the effect of tooth mousse containing CPP-ACP (GC Tooth Mousse®) or CPP-ACP with 0.2% fluoride (CPP-ACPF; GC Tooth Mousse Plus®; GCP) on S. mutans planktonic growth and biofilm formation.MethodsS. mutans was cultivated in the presence of different dilutions of the tooth mousse containing CPP-ACP or CPP-ACPF, and the planktonic growth was determined by ATP viability assay and counting colony-forming units (CFUs). The resulting biofilms were examined by crystal violet staining, MTT metabolic assay, confocal laser scanning microscopy (CLSM), and scanning electron microscope (SEM).ResultsThe CPP-ACP tooth mousse (GC) at a dilution of 5–50 mg/ml (0.5–5%) did not inhibit planktonic growth, and even increased the ATP content and the number of viable bacteria after a 24 h incubation. The same was observed for the CPP-ACPF tooth mousse (GCP), except for the higher concentrations (25 and 50 mg/ml) that led to a drop in the bacterial count. Importantly, both compounds significantly decreased S. mutans biofilm formation at dilutions as low as 1.5–3 mg/ml. 12.5 mg/ml GC and 6.25 mg/ml GCP inhibited biofilm formation by 90% after 4 h. After 24 h, the MBIC90 was 6.25 mg/ml for both. CLSM images confirmed the strong inhibitory effect GC and GCP had on biofilm formation when using 5 mg/ml tooth mousse. SEM images of those bacteria that managed to form biofilm in the presence of 5 mg/ml tooth mousse, showed alterations in the bacterial morphology, where the streptococci appear 25–30% shorter on the average than the control bacteria.ConclusionOur data show that the tooth mousse containing CPP-ACP reduces biofilm formation of the cariogenic bacterium S. mutans without killing the bacteria. The use of natural substances which inhibit biofilm development without killing the bacteria, has therapeutic benefits, especially in orthodontic pediatric patients.
Highlights
Streptococcus mutans is a common cariogenic bacterium in the oral cavity involved in plaque formation
The diluted casein phosphopeptides (CPPs)‐amorphous calcium phosphate (ACP) tooth mousse (GC) did not inhibit planktonic growth of S. mutans Our first question was whether the Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP)-containing tooth mousse affects the planktonic growth of the cariogenic S. mutans
We found a 1.4–3.3-fold increase in the bacterial count when S. mutans was exposed to 3–50 mg/ml GC tooth mousse containing CPP-ACP (GC) tooth mousse (Fig. 1b)
Summary
Streptococcus mutans is a common cariogenic bacterium in the oral cavity involved in plaque formation. The bacterium utilizes sucrose to synthesize adhesive fructans and glucans by the respective enzymes fructosyltransferase (FTF) and glucosyltransferase (GTF) [5]. These extracellular polysaccharides (EPS) stick to surfaces, such as the tooth enamel, and together with glucan-binding proteins (GBPs), they act as binding sites for S. mutans and other microbes, thereby forming dental plaques composed of a complex microbe community embedded in an extracellular matrix [5]. Besides contributing to dental plaque formation, S. mutans is highly cariogenic in virtue of its ability to process sucrose from nutritional substances into organic acids that lower the pH within the biofilm, resulting in decalcification of the tooth enamel [6]. The sessile state of the bacteria in the biofilm makes them less sensitive to anti-bacterial agents [7]
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