BACKGROUND:Bacteria contained in biofilms have been shown to have a decreased susceptibility to antimicrobial agents compared to those in planktonic form. Thus, in vitro biofilm models have been developed for screening oral antimicrobial formulations in an effort to produce findings more predictive of clinical activity. This study compared the antimicrobial activity of three mouthrinse formulations when tested against isogenic strains of Actinobacillus actinomycetemcomitans (Aa), one of which was a clinical isolate which forms tenacious biofilms in vitro and the other of which was a spontaneous variant which always grows planktonically.METHOD:Biofilm-forming Aa strains CU1000 and NJ4300, obtained as clinical isolates, and their respective spontaneous planktonic variants, CU1060 and NJ4350, were grown under standard laboratory conditions and exposed for 15 s to either a negative control (phosphate buffered saline [PBS]), an essential-oil containing mouthrinse (Listerine Antiseptic [LA]), an amine fluoride/stannous fluoride-containing mouthrinse (Meridol [M]), or a triclosan and PVM/MA copolymer-containing mouthrinse (Plax [P]). The cells were then washed, serially diluted, plated, and incubated for enumeration of viable bacteria. Colony-forming units (CFU)/ml were log10 transformed and the mouthrinse groups were compared to the PBS group using analysis of variance.RESULTS:All 3 mouthrinses produced statisically significant 99.99% reductions (p< or =0.0001) in both planktonic strains compared to the PBS control. Effects on the biofilm forms of the organisms were more variable. Exposure to LA produced statistically significant (p< or =0.0001) reductions in strains CU1000 and NJ4300 of 98.20% and 96.47%, respectively, compared to PBS. M and P produced much smaller reductions which were not statistically significant.CONCLUSIONS:The results of this study, in which antimicrobial mouthrinses were tested against biofilm-forming and planktonic strains of the same organism, provide a clear demonstration of the resistance to antimicrobial agents conferred by biofilm formation and provide additional support for employing tests using biofilms to more accurately assess the relative activities of antiplaque agents in vitro.

BACKGROUND:Clinical studies in which antimicrobial mouthrinses were shown to have significant antiplaque activity most frequently have used gingivitis as the clinically relevant endpoint. However, there is evidence to suggest that mouthrinses containing active agents effective against Streptococcus mutans, such as chlorhexidine, may also have a role in inhibiting dental caries. This clinical study was conducted to determine the effect of 2x-daily rinsing with an essential oil-containing antiseptic mouthrinse (Listerine Antiseptic) on levels of recoverable S. mutans and total streptococci in supragingival interproximal plaque and in saliva. Additionally, a follow-up in vitro study is reported which determined whether a differential susceptibility to the antiseptic mouthrinse exists among different strains of streptococci.METHOD:Following baseline saliva and plaque sampling for quantification of recoverable S. mutans and total streptococci, 29 qualifying subjects were randomly assigned either the essential oil mouthrinse or a sterile water control. They rinsed with 20 ml for 30s 2 x daily for 11 days and once on the 12th day, in addition to their usual oral hygiene procedures. On day 12, saliva and plaque samples were again collected and microbiological quantification performed. The procedures were repeated with the alternate rinse after a 1-week washout period.RESULTS:The essential oil mouthrinse produced respective reductions of 69.9% and 75.4% in total recoverable streptococci and in S. mutans in plaque, and corresponding reductions of 50.8% and 39.2% in saliva. The in vitro study revealed that streptococci from the mutans group were more susceptible to the bactericidal activity of the essential oil mouthrinse than streptococci from the mitis group.CONCLUSIONS:As antimicrobial mouthrinses are most frequently recommended to patients whose mechanical oral hygiene procedures are not adequate for the control of supragingival plaque and gingivitis, this study provides an additional rationale for the inclusion of the essential-oil mouthrinse as an adjunct to daily oral hygiene procedures.

The adjunctive use of antimicrobial mouthrinses to help control supragingival plaque and gingivitis has been shown to contribute significantly to patients' daily oral hygiene regimens. This controlled clinical study used an observer-blind, randomized, cross-over design in a 4-day plaque regrowth model to determine the relative efficacies of an essential oil-containing mouthrinse (Listerine(R) Antiseptic) and an amine fluoride/stannous fluoride-containing mouthrinse (Meridol(R)) in inhibiting the development of supragingival plaque. A 0.1% chlorhexidine mouthrinse (Chlorhexamed(R)-Fluid) was used as a positive control, and a 5% hydroalcohol solution was used as a negative control. Dosing for each of the test mouthrinses was based on the manufacturers' label directions. Because the volume and rinse time for each of the lest mouthrinses were different, each test mouthrinse had its own negative control group. On day 1 of each test period, subjects received an oral soft and hard tissue examination and a dental prophylaxis to remove all plaque, calculus, and extrinsic stain. Starting the same day, subjects refrained from all mechanical oral hygiene procedures for the next 4 days and rinsed 2x daily under supervision with their randomly-assigned mouthrinse. On day 5, each subject received a plaque assessment as well as an oral examination to assess side effects. Each lest period was separated by a 2-week washout period. 23 volunteers with a median age of 26 years completed the study. Compared to the respective placebos, the median percent plaque reductions at 5 days were 23.0%, 12.2%, and 38.2% for the essential oil, amine/stannous fluoride, and chlorhexidine rinses, respectively. The plaque reductions seen in the essential oil and chlorhexidine rinse groups were statistically significant (p<0.001), while the plaque reduction in the amine/stannous fluoride rinse group was not statistically significant (p>0.05). Additionally, the essential oil rinse was significantly more effective (p<0.001) than the amine/stannous fluoride rinse in inhibiting plaque accumulation in this clinical model.

In this controlled clinical study, the authors examined the effect of subgingival irrigation and rinsing With an antiseptic mouthrinse before ultrasonic scaling of a quadrant containing inflamed gingivae. The results showed that pre-procedural subgingival irrigation and rinsing can significantly reduce the level of bacteremia associated with ultrasonic scaling. These results support the American Heart Association's recommendation of adjunctive subgingival irrigation prior to invasive procedures in patients at risk of developing bacterial endocarditis.

Inthiscontrolled clinical study, theauthorsexaminedtheeffect ofsubgingival irrigation and rins- ingwithan antiseptic mouth- rinsebeforeultrasonic scalingof a quadrantcontaining Inflamed gingivae. The resultsshowed thatpre-procedural subgingival irrigation and rinsing can signifi- cantlyreducethelevelofbac- teremiaassociatedwithultra- sonicscaling. Theseresults supporttheAmericanHeart Association's recommendation ofadjunctive subgingival irriga- tionprior toInvasive procedures inpatients atriskofdeveloping bacterial endocarditis.