Comparison of the efficacy of three different mouthrinse solutions in decreasing the level of streptococcus mutans in saliva.

OBJECTIVES: The aim of this study was to evaluate the effectiveness of three different antiseptic mouthrinse solutions on the saliva samples obtained from the individuals, who had high caries activity rate.
METHODS: The efficacy of three antiseptic mouthrinses were evaluated in a study with healthy volunteers. The three antiseptic solutions used in this study were 0.1% octenidine dihydrochloride (Octenisept, Schülke&Mayr, UK), 0.12% chlorhexidine digluconate (Kloroben, Drogsan, Turkey) and an antimicrobial enzymatic rinse (Biotene, Laclede, Inc, USA). A total of 27 adult volunteer subjects were participated in the study. The subjects were stratified into three balanced group. Then the mouth rinses were used by each group according to the manufacturer's directions. The subjects were restricted for 60 minutes for food intake after using the prescribed mouthrinse. The saliva samples were collected from the volunteers at 1, 10 and 60 minutes after their usage in tubes. The tubes were kept in +4 degrees C in a fridge till the evaluation. 10(-3) and 10(-5) dilutions were prepared for each solution and S. mutans were evaluated according to total number of colony forming unit (CFU) per ml. The dilutions were spreaded on the surface of Brucella agar plates for anaerobic incubation for 48 hours. The dilutions were 100, 10(-3) and 10(-5) of the solutions Kloroben, Biotene, Octenisept, and the time factor were 0, 1, 10 and 60 minutes. The statistical analyses were performed by Duncan and Bonferroni tests.
RESULTS: Octenisept was found to be more effective over S. mutans than the other mouthrinse solutions (P<.05).
CONCLUSIONS: All mouthrinse solutions except Biotene were effective on oral microorganisms.

INTRODUCTION

Antiseptic mouth rinse solutions are used in many clinical situations for different prophylactic and therapeutic purposes. It is difficult to decide which product is suitable for a particular purpose because of the variations of the antimicrobial efficacy and kinetics of the solutions.1 The main indications are either the improvement of dental health (plaque and gingivitis elimination in particular) or the prevention of infections caused by bacteria of the oral cavity in specific situations such as tooth extraction, intraoral surgical procedures or immuno suppression due to cancer therapy or transplantation.2,3 The use of antimicrobial mouth rinses has been proposed as a means of reducing the levels of oral bacteria, specifically Streptococcus mutans (S. mutans).4

Chlorhexidine is a broad-spectrum antimicrobial agent. In dentistry, chlorhexidine has been used to reduce the level of oral S. mutans, and incorporated into mouth rinse solutions. Chlorhexidine was shown to inhibit plaque formation, also reduce gingival inflammation and prevent dental caries.5 However, studies aimed at reducing the levels of S. mutans in the oral cavity with chlorhexidine had reported large variations, inconsistencies, and an inability to ablate S. mutans.6

Octenidine dihydrochloride was developed at the Sterling Winthrop Research Institute as a potential topical antimicrobial agent.7 In a previous study this compound was found to be effective in inhibiting the growth of plaque forming bacteria8,9 and in reducing the development of plaque in experimental animals.10

Biotene Mouthwash (Laclede, CA, USA) is especially beneficial to individuals experiencing dry mouth or having oral irritations. The strength of the Biotene products lies in their ingredients: they contain antibacterial enzymes which found naturally in human saliva. Biotene contains three primary enzymes-Glucose Oxidase, Lactoperoxidase, and Lysozyme, which are carefully balanced for a special function in boosting and replenishing saliva’s own defenses. These ingredients’ antibacterial and healing properties create a natural oral protection.11

The aim of this study was to evaluate the effectiveness of three different antiseptic mouth rinses on the saliva samples of individuals, who had high caries activity rate.

MATERIALS AND METHODS

A total of twenty-seven, fourteen male and thirteen female, healthy volunteers aged between 20–22 were included in the study. The participants had at least four restored teeth in their mouth but without any decay or tooth lose. Antibiotic or other medication consumption in the last two months that might interfere oral hygiene and participants with periodontal problems and the ones who were taking special diet were excluded from the study. First, the medical stories of the volunteers were taken and they were requested to sign a consent form, afterwards all of them received professional tooth cleaning. Then a participant number was given for each and randomly divided into three equal groups. The volunteers In Group 1, rinsed with Octenisept (Octenidine dihydrochloride), in Group 2 with Kloroben (Chlorhexidine digluconate) and in Group 3 with Biotene. The groups rinsed for two minutes with the mouth rinses (Table 1) according to the manufacturers’ instructions. The saliva samples were collected with sterile tubes at the 1, 10 and 60 minutes following the rinsing procedure. The tubes were kept in +4°C in a fridge till the evaluation. The solutions were diluted with distilled water. The dilutions were spreaded on the surface of Brucella agar plates for anaerobic incubation for 48 hours. The dilutions were 100, 10−3 and 10−5 of the solutions Kloroben, Biotene, Octenisept, and the time factor were 0, 1, 10 and 60 minutes. S. mutans were evaluated according to total number of colony forming unit (CFU) per ml.

Table 1

Mouth rinse solutions.

Product	Listed ingredients	Manufacturer
Octenisept	% 0.1 Octenidine dihydrochloride % 2 2-Phenoxyethanol	Schülke & Mayr Sheffield, UK
Kloroben	% 0.12 Chlorhexidine digluconate % 0.15 Benzidamin HCI	Drogsan Ankara, Turkey
Biotene	Enzyme system, distilled sterile water, Propylene Glycol, Xylitol, Sodiumbenzoat, Benzoic acid, Aloe vera, Calcium lactate, Potasium tiocionate	Laclede Inc. CA, USA

The data were collected and statistical analysis were performed by Duncan and Benferroni tests. Also factorial repeated ANOVA was used for numerical evaluation of bacteria.

RESULTS

There were no S. mutans growth in Octenisept group at 1 and 10 minute calculations for all dilution levels (100, 10−3 and 10−5), but bacterial growth was observed only in one specimen at 60 minute group and when compared with the initial values a statistically significant difference was found (P<.05). In Kloroben group for all dilution levels (100, 10−3 and 10−5) at 1 and 10 minute calculations there was a significant reduction in S. mutans amount when compared with the initial values and this reduction was also statistically significant (P<.05). At the 60 minute calculations of the same group, no significant difference was found (P<.05). In Biotene group, for all dilution levels and time periods (for 1, 10 and 60 minutes calculations) no statistically significant difference was found (P<.05).

In Table 2 the mean differences and the standart deviations of the rinsing solutions were given in latin letters. No differences were found between the groups stated with the same letters, but the statistical differences are clear between the groups stated with different letters.

Table 2

Statistical results of the mouth rinse solutions.

Solutions	x ± S x
Kloroben	2.33 ± 0.107 A
Biotene	2.67 ± 0.871 A
Octenisept	1.00 ± 0.261 B

(P<.05)

DISCUSSION

The oral cavity represents a dynamic ecosystem therefore it would not be totally advantageous to eliminate all elements of the oral microflora in an effort to control dental plaque-associated infections. Rather, it may be more ideal to remove only most cariogenic and periodontopathic elements of the dental plaque microflora while permitting the more innocuous elements to remain.8

A relatively large number of chemical agents, which are mostly synthetic compounds, have been used for many purposes, control of dental plaque, elimination of oral pathogens, against malodor, etc.12

The presented study was designed for the evaluation of the antimicrobial effects of a new solution containing octenidine dihydrochloride (Octenisept), a chlorhexidine-based product (Kloroben) and a mouth rinse which contains natural antimicrobial enzymes (Biotene) on the the levels of S. mutans in saliva.

Octenidine dihydrochloride (OCT) was originally developed as a potential broadspectrum topical antimicrobial agent13 and used as an oral rinse is reported to inhibit dental plaque and caries in rats,14 dental plaque in-primates15 and in humans.16 One of the recent studies showed that a 0.1% octenidine mouth rinse provided statistically significant reductions of 39% less plaque, 50% less gingivitis, and 60% fewer gingival bleeding sites.17 In this study, it was observed that Octenisept had a significant effect on S. mutans and preserved antimicrobial efficiancy even after 60 minutes.

Chlorhexidine (CHX) digluconate has a 30-year history in dental medicine.18 It is the most throughly studied and the most effective anti-plaque and anti-gingivitis agent known today.19 Gjermo et al20 reported that rinsing twice a day with 10 ml of a 0.2% CHX inhibited the dental plaque formation. Furthermore, its anti-gingivitis efficacy was also well documented.21–23 Unfortunately, these positive effects are accompanied by side effects, the most disturbing being extrinsic tooth staining.24–27 In few cases, the occurrence of gingival desquamation and painful mucosa were reported.23,25

In our study, Kloroben had significantly reduced the S. mutans levels in saliva samples. However, it was observed that its efficiency was lower than Octenisept in the 60th minute. Also Robrish et al28 reported that OCT had a more persistent antimicrobial effect on the organisms in plaque than that obtained by CHX. However, Dogan et al29 compared the short-term relative antibacterial effects of OCT and CHX. Their results were similar with our study, OCT was found favorably more effective than CHX in its antibacterial activity, both in vitro and in vivo. Although the initial antimicrobial activities of OCT and CHX are comparable, as a result of its better persistant antimicrobial activity we may suggest that OCT have promising effeciency on S. mutans as a mouth rinse solution.

Although the results showed that Biotene had no effects on salivary S. mutans levels, further studies are required to observe the effects of the solution on the oral flora and the oral cavity.30

CONCLUSIONS

The finding of the present study suggest that OCT and CHX mouthrinses are extremely effective in reducing S. mutans levels in saliva. As a result, the data presented in this study allows classification of different mouthrinse solutions due to their efficacy in decreasing the levels of S. mutans in saliva and enable the prescribing dentist or oral hygienist to make his choice based on antimicrobial impact.