ABSTRACT
The aim of the present study was to formulate toothpastes containing biosurfactants and either fungal chitosan or sodium fluoride and evaluate the cytotoxicity, antimicrobial action and inhibition potential against biofilm formed by Streptococcus mutans. Chitosan was extracted from the biomass of the fungus Mucorales. We tested biosurfactants produced by Pseudomonas aeruginosa UCP 0992 (PB), Bacillus metylotrophicus UCP 1616 (BB) and Candida bombicola URM 3718 (CB). Fractional inhibitory concentration analysis was performed to determine the type of interaction between the compounds. Six toothpaste were prepared, the active ingredients of which were the biosurfactants, chitosan or sodium fluoride. The cytotoxicity tests were performed using the 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay for the L929 (mouse fibroblast) and RAW 264.7 (mouse macrophage) cell lines. The toothpastes were tested with regard to pH, consistency and foaming capacity. The inhibition of biofilm was investigated by applying the toothpaste to biofilm formed in modified artificial saliva for 24 h at 37 °C in anaerobiosis. All substances had a minimum inhibitory concentration (MIC) for S. mutans. The combinations of CB and PB with chitosan had an additive effect against S. mutans, whereas BB combined with chitosan had an indifferent effect. The toothpastes were non-toxic. The formulations had pH around 9, spreading capacity between 8 and 17 mm and foaming capacity between 63 and 95%. All formulations inhibited the cellular viability of S. mutans in the biofilm, with similar results compared to the commercial toothpaste tested. The present results show that the formulations suggested are promising when compared to a commercial tooth paste.
Subject(s)
Chitosan/pharmacology , Streptococcus mutans/drug effects , Surface-Active Agents/pharmacology , Toothpastes/chemistry , Toothpastes/pharmacology , Animals , Biofilms/drug effects , Cell Line , Cell Survival/drug effects , Chitosan/chemistry , Chitosan/isolation & purification , Fibroblasts/drug effects , Mice , Particle Size , RAW 264.7 Cells , Sodium Fluoride/chemistry , Sodium Fluoride/isolation & purification , Sodium Fluoride/pharmacology , Surface Properties , Surface-Active Agents/chemistry , Surface-Active Agents/isolation & purification , Toothpastes/isolation & purificationABSTRACT
The aim of the present study was to determine the antimicrobial action and toxicity of mouthwashes formulated with a biosurfactant, chitosan of a microbial origin and peppermint (Mentha piperita) essential oil (POE). Chitosan was extracted from the biomass of a fungus from the order Mucorales grown in yam bean broth. Three biosurfactants produced by Pseudomonas aeruginosa UCP 0992 (PB), Bacillus cereus UCP 1615 (BB) and Candida bombicola URM 3718 (CB) were tested. Six mouthwashes were prepared, the active ingredients of which were the biosurfactant, chitosan and POE. The minimum inhibitory concentration (MIC) was determined for the test substances separately, in combinations and in the mouthwash formulas. The toxicity of the mouthwashes was tested using MTT (3-(4,5-dimethylthiazole-2-il)-2,5-diphenyltetrazolium bromide) for the L929 (mouse fibroblast) and RAW 264.7 (mouse macrophage) cell lines. All substances tested had a MIC for cariogenic microorganisms. The combinations of the CB and PB biosurfactants with chitosan demonstrated an additive effect on the majority of microorganisms tested. The toxicity of the mouthwashes was significantly lower than that of the commercial mouthwash. The present findings demonstrate that mouthwashes containing natural products constitute a safe, effective, natural alternative to commercially available mouthwashes for the control of oral microorganisms.
