Effect of Ozonated-Water on in-situ Plaque and Whole Saliva

Objectives: To determine the in-situ antibacterial effect of 0.1 ppm ozonated-water on plaque microorganisms and salivary calcium and protein. Methods: The study was carried out as double blind, randomised study. Forty healthy volunteers were randomly divided into two groups (Group1-control, Group2-test) of 20 individuals. They were instructed not to brush their teeth 24 hours prior to sample collection. Pre-rinse ‘stimulated whole saliva' and ‘24-hour supragingival plaque' samples were collected from the buccal and lingual surfaces of the three most posterior teeth on the upper right and lower left quadrants of both the groups. Following that Group1 was made to gargle with 20 ml distilled water and Group2 with 20 ml 0.1ppm ozonated-water for 30 seconds. 20 minutes post-rinse saliva and plaque samples were collected from the three most posterior teeth on the upper left and lower right quadrants. The total-microbial, anaerobic, streptococcal and candidal concentrations of plaque were quantitatively determined based on the growth of colony forming units (cfu) on selective media plates. Microbial concentration of plaque and saliva were also qualitatively assessed using fluorescence staining microscopy. Salivary calcium and protein content were determined by titration and spectrophotometry respectively. Results: Results showed a significant lower total-microbial and anaerobic load in the post-rinse samples of ozonated-water treated group compared to the distilled water treated group (p<0.01 and p<0.02). There was an average reduction of 45.35% (SD±26.24) of total bacteria and 51.71% (SD±18.79) of anaerobic bacteria observed in Group2. These findings positively correlated with fluorescence staining results. No significant changes in the concentration of salivary calcium and protein were observed for both groups. Conclusion: Gargling with 0.1ppm ozonated-water has shown statistically significant antimicrobial effect on plaque microorganisms but did not induce changes to the protein and calcium content of saliva. This study was supported by VoteF F0144/2005A -University Malaya.