|Year : 2012 | Volume
| Issue : 1 | Page : 39-42
|Efficacy of commercial and household denture cleansers against Candida albicans adherent to acrylic denture base resin: An in vitro study
M Nanditha Kumar1, HM Thippeswamy2, KN Raghavendra Swamy3, Anil Kumar Gujjari3
1 Department of Prosthodontics, Bapuji Dental College and Hospital, Davangere, India
2 Department of Prosthodontics, Public Health Dentistry, Bapuji Dental College and Hospital, Davanagere, India
3 Department of Prosthodontics, J.S.S. Dental College and Hospital, Mysore, Karnataka, India
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|Date of Submission||16-Jul-2010|
|Date of Decision||15-Apr-2011|
|Date of Acceptance||08-Nov-2011|
|Date of Web Publication||26-Jul-2012|
| Abstract|| |
Aim: To evaluate and compare the efficacy of two commercial and two household denture cleansers against Candida albicans adherent to acrylic denture base resin.
Materials and Methods: Fifty specimens of acrylic denture base resin (10 × 10 × 2 mm) were fabricated and processed according to the manufacturer's instructions. Sterile acrylic resin specimens were inoculated by immersing in Sabouraud broth containing C albicans for 16 hours at 37°C in an incubator. Then the specimens were washed and immersed in denture cleansers (four groups) - Clinsodent® (powder form), Fittydent® (tablet form), vinegar (4% acetic acid), diluted vinegar (50% diluted with water), and water (control group) for 8 hours at room temperature. After 8 hours the acrylic resin specimens were washed, fixed with methanol, and stained with crystal violet. Candida cells adherent to the acrylic resin specimens were counted under microscope. The number of cells adherent to the test samples were compared with that adherent to the control.
Statistical Analysis: The data were analyzed using the independent-samples 't ' tests, analysis of variance (ANOVA), and Tukey's HSD test.
Results: All the denture cleansers were highly effective against C albicans. The effectiveness of commercial denture cleansers was significant better than that of household denture cleansers. Fittydent® fared better than Clinsodent® , but the difference between the two was not statistically significant (P=.765). Vinegar was more effective than diluted vinegar (P<.05).
Conclusion: Within the limitations of this in vitro study, denture cleansers were found to be effective in reducing C albicans cells adhering to dentures. The commercial denture cleansers (Fittydent® and Clinsodent® ) were more effective than household denture cleansers (vinegar and diluted vinegar).
Keywords: Acrylic resin, Candida albicans, commercial denture cleansers, household denture cleansers
|How to cite this article:|
Kumar M N, Thippeswamy H M, Raghavendra Swamy K N, Gujjari AK. Efficacy of commercial and household denture cleansers against Candida albicans adherent to acrylic denture base resin: An in vitro study. Indian J Dent Res 2012;23:39-42
The increase in the number of dentures in service has stimulated interest in measures to control the deposits and stains that normally collect on them. The use of teeth with more natural contours and the trend toward stippled surfaces tend to provide more recessed areas for the accumulation of plaque, stain, and debris and consequently increases the patient's cleaning problems.  Denture cleanliness is essential to prevent malodor, poor esthetics, and accumulation of plaque/calculus and consequent deleterious effects on the mucosa. 
|How to cite this URL:|
Kumar M N, Thippeswamy H M, Raghavendra Swamy K N, Gujjari AK. Efficacy of commercial and household denture cleansers against Candida albicans adherent to acrylic denture base resin: An in vitro study. Indian J Dent Res [serial online] 2012 [cited 2015 Mar 2];23:39-42. Available from: http://www.ijdr.in/text.asp?2012/23/1/39/99036
Denture-induced stomatitis is the pathogenic reaction of denture-bearing mucosa and one of the most common states of chronic candidosis.  Candida albicans in the microbial plaque on denture fitting surface is thought to be an important etiologic factor in the pathogenesis of denture-induced stomatitis, ,,,, which only goes to show that the treatment of the denture is as important as treatment of the tissue. 
Dentures can be cleaned mechanically, chemically, or by a combination of the two.  Mechanical cleansing methods like brushing are effective when carried out meticulously but it can also abrade the denture surface. Chemical agents, besides being simple to use, are effective in reducing candidal biofilm formation.  Hence, chemical denture cleansers are commonly prescribed alternatives to mechanical cleaning.
Although many commercial chemical denture cleansers are available, patients also often use household cleansers because of its ease of availability and low cost. The popular household cleansers are vinegar, diluted vinegar, soda, salt, and bleach.  Acetic acid is one component of vinegar. Lately, interest in vinegar and other solutions of acetic acid as an antimicrobial solution has increased. 
There have not been many studies comparing commercial and household cleansers and therefore the antimicrobial effect of household denture cleansers in not well understood. The aim of this study was to evaluate the efficacy of commercial and household denture cleansers against C albicans adherent to acrylic denture base resin.
| Materials and Methods|| |
Fabrication of acrylic resin specimen
Fifty denture base acrylic resin specimens with the same dimensions (10 × 10 × 2 mm) were obtained by means of wax patterns. A metal mold with three plates was fabricated. Wax (modeling wax, Hindustan Dental Products) was melted and poured into the openings, measuring 10 × 10 × 2 mm each, in the middle plate of the metal mold.  After the wax hardened, the wax patterns were numbered according to the groups. The wax patterns were flasked according to conventional technique and dewaxing was carried out. Acrylic resin (DPI Heat-Cure Improved; DPI India, India) was packed and trial and final closures were done. After bench curing for 30 minutes, polymerization was carried out according to the manufacturer's instruction. The flasks were allowed to cool to room temperature before being opened. Finishing was done on one of the surfaces using fine-grit sandpaper and polished on a wet rag wheel with pumice slurry. The fifty specimens were divided into five groups, with ten specimens in each group.
Sterilization of acrylic resin specimen
The acrylic resin specimens were sterilized in ultraviolet light (wavelength: 250 nm) chamber (Alpha Scientific, Bangalore, India) for 5 minutes.  As ultraviolet light does not penetrate opaque materials, the specimens were overturned using sterile forceps and the other side was sterilized for 5 min. The acrylic specimens were then placed in a zip-lock plastic bag (Uflex Ltd., Nodia, India), which was sterilized in the same ultraviolet light chamber.
Preparation of broth
Two to three colonies of C albicans grown on blood agar was taken and inoculated onto Sabouraud dextrose broth. This was incubated aerobically at 37°C for 24 hours and the density of the broth was matched to 1 McFarland unit (10 6 CFU/ml).
Preparation of denture cleanser solutions
Denture cleanser solutions for the different groups were prepared as follows:
- Addition of 1 tablet of Fittydent® (Peroxide cleanser, Group Pharmaceuticals Ltd, India) to 100 ml of water
- Addition of one teaspoon-full powder of Clinsodent® (peroxide cleanser, ICPA Health Products, India) to 100 ml of water
- Vinegar (4% acetic acid, A.W. Wilson and Co., India) for use as such
- Vinegar diluted with an equal amount of water
- Plain water (for the control group).
Exposure of specimens to C albicans
The sterile acrylic resin specimens of various groups were immersed, with the labeled surfaces facing down, in Petri dish More Detailses containing 20 ml of Sabouraud dextrose broth containing C albicans. These were then incubated for 16 hours at 37°C in incubator. This simulated the duration of the denture worn by the patient during the daytime. Then the inoculated specimens were washed under running tap water to simulate the patient's routine denture cleaning procedure.
Candida removal test
The specimens were immersed in petri dishes containing 50 ml of denture cleansers - Clinsodent® , Fittydent® , vinegar, diluted vinegar, and water (control). They were stored for 8 hours at room temperature. This mimicked the overnight soaking of dentures in cleansers according to the manufacturers' instructions. After this, the specimens were washed under running tap water, fixed with methanol, stained with crystal violet, dried, and examined under the microscope. 
Counting of Candida cells
Candida cells adherent to the acrylic resin specimens were counted under the microscope (×40 magnification). The entire surface (10 × 10 mm) of the specimen was counted. Each field (1.1 mm 2 ) was counted and totaled. The number of cells adherent on the test samples were compared with that adherent to the control.
Differences in the percentage of Candida cells were analyzed using descriptive statistics, independent-samples 't' test, and ANOVA. All the statistical calculations were done using SPSS® (Statistical Package for the Social Sciences) for Windows® version 14.0 (SPSS Inc, New York).
| Results|| |
This study was done primarily to evaluate and compare the effectiveness of commercial and household denture cleansers. The results are shown in [Table 1]. The effectiveness of Fittydent® and Clinsodent® against C albicans were almost the same; Fittydent® was slightly more effective than Clinsodent® In reducing adherent Candida but the difference was not statistically significant (P=.765).
|Table 1: Comparison of commercial and household chemical denture cleansers|
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Both vinegar and diluted vinegar were found to be significantly better than control (P<.05) against C albicans. Vinegar was more effective than the diluted vinegar.
Post hoc Tukey's analysis indicates that Fittydent® was the most effective among the denture cleansers tested in this study, followed by Clinsodent® , vinegar, and diluted vinegar.
| Discussion|| |
The increased awareness of patients regarding dentistry and about the relationship between poor oral hygiene and systemic diseases has stimulated interest in patients in improving the oral hygiene status. In geriatric patients, infections of the oral cavity by various microorganisms is a matter of major concern.  Drake et al. found that denture plaque containing C albicans plays a major role in the pathogenesis of denture stomatitis.
A routine denture cleansing regimen should be followed to remove and prevent re-accumulation of plaque. Denture cleansing may be carried out using mechanical methods, chemical methods, or a combination of the two. According to Jagger and Harrison,  mechanical methods used overenthusiastically or with an incorrect brushing technique can abrade the denture base material. Besides mechanical methods are also of limited value in individuals with impaired manual dexterity.
Chemical cleansing methods have proved to be more efficient than mechanical methods as suggested by Sato et al.  Chemical agents for denture cleansing have the advantage of being simple to usem, and several investigations have shown their efficacy in reducing biofilm formation in vitro and in vivo. In vitro studies help in understanding the mode of action of denture cleansers under clinical conditions. 
The main aim of this in vitro study was to evaluate and compare the effect of commercially available and common household denture cleansers on adherent Candida cells. According to Nakamoto et al. chemical denture cleansers can be divided into five groups based on their main components as: Alkaline peroxides, alkaline hypochlorites, acids, disinfectants, and enzymes. Among these, alkaline peroxides are the most commonly used. , Therefore, two alkaline peroxide cleansers - Fittydent® (tablets) and Clinsodent® (powder) - were chosen for this study.
For geriatric patients, cost and easy availability are important factors when selecting a denture cleanser. Anthony and Gibbons,  have suggested that household products like vinegar, which is inexpensive and easily available, can be used as denture cleansers. Hence, vinegar and diluted vinegar were selected for this study. Tap water was used as the control.
The removal of adherent cells from the acrylic resin surface is an important part of denture cleansing. The Candida removal test is a quantitative measure of the efficiency of a denture cleanser.  As such, we conducted the Candida removal test to check the efficacy of the denture cleaning agents. The present study evaluated and compared the effectiveness of different denture cleansers in removing Candida from the surface of acrylic resin specimen.
The Candida removal test revealed that the commercial denture cleansers (alkaline peroxide cleansers) were better than household denture cleansers (acid cleansers). Fittydent® removed C albicans from acrylic resin surface better than the other denture cleansers after immersion of the specimens for 8 hours. Clinsodent® and vinegar were second and third, respectively, in efficiency. Diluted vinegar was the least effective among the denture cleansers. This can be explained by the fact that, the primary means of adherence of plaque is through its organic portion. Alkaline peroxides when dissolved in water form solutions of hydrogen peroxide and liberate nascent oxygen in the presence of organic material. The oxygen bubbles exert a mechanical cleansing effect. Acids dissolve calcareous deposits and denature the proteins. Thus, alkaline peroxide cleansers, which act on the organic portion of plaque, are able to remove Candida from acrylic surface better than acid cleansers, which act mainly on the inorganic portion of denture plaque.
Among the commercial denture cleansers Fittydent® and Clinsodent® were equally effective in the removal of Candida cells from the acrylic resin surface. Among the household denture cleansers, vinegar was more effective than diluted vinegar. These results are in agreement with the study done by Pinto et al. They too demonstrated significant reduction in Candida count after immersion in 10% vinegar solution.
This study was carried out on the clinically relevant C albicans. The study also simulated the patient's routine denture cleansing procedure. However, it is difficult to extrapolate our findings to the in vivo situation as the study was carried out under laboratory conditions. Also, this study simulated only one day of the denture cleansing procedure. Studies simulating the denture cleansing protocol for longer periods of time would yield more accurate information. None of the treatments were able to completely remove Candida cells, which means that these cleansers alone are not sufficient. Re-colonization may occur as some Candida cells remain on the surface. Further research is needed to test the efficiency of denture cleansers on various microorganisms.
| Conclusion|| |
A simple method was employed to assess the efficacy of denture cleansers against C albicans and, within the limitations of this in vitro study, the authors conclude:
- The commercial denture cleansers (Fittydent® and Clinsodent® ) were more effective than household denture cleansers (vinegar and diluted vinegar) in removing C albicans from the acrylic specimen after immersion for 8 hours.
- The commercial denture cleansers (Fittydent® and Clinsodent® ) were equally effective against C albicans.
- Among the household denture cleansers, vinegar was more effective in reducing the number of Candida cells than diluted vinegar.
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M Nanditha Kumar
Department of Prosthodontics, Bapuji Dental College and Hospital, Davangere
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