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1174 non-alcoholic calculus dissolution-based oral rinse containing tetrapotassium pyrophosphate (TKPP), sodium tripolyphosphate (STPP), sodium bicarbonate, sodium fluoride and citric acid (Periogen®, USA). The positive control was a 0.12% CHX solution, while the negative control was a physiologic saline solution (PSS)(placebono mouthwash dilution added). All bottles with mouthwash provided to subMects were pre-weighed. All participants were instructed to refrain from using any other means of oral hygiene during the experimental period. All subMects were instructed to rinse twice a day, in the morning and in the evening, with an applicator spoonful dissolved in one cup of warm water for 60 seconds, or 20 ml of solution for CHX and PSS. Subsequent rinsing with water was not allowed and the subMects were also asked not to brush their teeth during the study period. For plaque scores, the participants rinsed for 30 seconds using a disclosing solution (1% erythrosine), according to Quigley and Hein index, Turesky modification (13). All measurements were carried out under similar clinical settings and by the same blinded investigator. The plaque and bleeding scoring were recorded prior to saliva collection. All returned mouthwash bottles were weighed to calculate the amount of mouthrinse used and to check for compliance.

In order to have microbial samples representative of health-and disease-associated conditions, the 22 volunteers selected included 11 periodontally healthy individuals (less than 10% of the sites with bleeding of gingival margin, plus a maximum of 2 sites with probing depth> 6.0 mm or attachment level> 5.0 mm …