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 Table of Contents  
ORIGINAL ARTICLE
Year : 2017  |  Volume : 10  |  Issue : 2  |  Page : 196-202

Comparative evaluation of the effect of commercially available two different forms of denture cleansers on denture biofilm in diabetic and nondiabetic individuals: An in vivo study


1 Department of Prosthodontics, K. L. E. V. K. Institute of Dental Sciences, Belagavi, Karnataka, India
2 Department of Microbiology, J.N.M.C. Medical College, Belagavi, Karnataka, India

Date of Web Publication30-May-2017

Correspondence Address:
Heena Sharma
Department of Prosthodontics, K. L. E. V. K. Institute of Dental Sciences, Belagavi, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/kleuhsj.ijhs_434_16

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  Abstract 

Statement of Problem: Denture cleansers are known to decrease the microbial counts in denture wearers. In certain medical conditions such as in diabetes, achieving reduction in microbial count and maintaining proper denture hygiene is difficult. Denture cleansers are used as an aid in maintaining denture hygiene in such patients. A number of tablets, pastes, and powders are available for cleaning dentures. Although the comparison of chemical methods and brushing is relatively common, their difference in diabetic denture biofilm and nondiabetic denture biofilm has not been previously described.
Purpose: The purpose of this study was to investigate the effect of commercially available two different forms of denture cleansers on the denture biofilm in diabetic and nondiabetic denture-wearing individuals.
Methodology: Twenty diabetic denture and 20 nondiabetic denture wearers were randomly selected for the study. Denture biofilm was collected twice for each participant, before and after use of denture cleanser for 21-day period. For sample collection, complete dentures were rinsed with distilled water and brushed. Inoculation was done with sterile loop and cultured on blood agar and chrome agar. Blood agar plates were incubated at 370°C for 24 h and chrome agar at 290°C for 24 h. Colonies' characteristics of the Streptococcus species and Candida species were counted after the incubation period.
Results: The results showed a statistically significant reduction in the colony-forming unit (CFU) counts after the usage of denture cleansers. Furthermore, the Candidal count difference obtained by the use of effervescent tablets was more than paste form.
Conclusions: The present study concluded that microbial colonization count of diabetic and nondiabetic denture biofilm significantly reduced after 21 days of usage of denture cleansers, and the effervescent tablets were more effective against Candida than paste form.

Keywords: Denture cleansers, diabetes, microbial colonization


How to cite this article:
Sharma H, Patil R, Nagmoti JM. Comparative evaluation of the effect of commercially available two different forms of denture cleansers on denture biofilm in diabetic and nondiabetic individuals: An in vivo study. Indian J Health Sci Biomed Res 2017;10:196-202

How to cite this URL:
Sharma H, Patil R, Nagmoti JM. Comparative evaluation of the effect of commercially available two different forms of denture cleansers on denture biofilm in diabetic and nondiabetic individuals: An in vivo study. Indian J Health Sci Biomed Res [serial online] 2017 [cited 2020 Jul 6];10:196-202. Available from: http://www.ijournalhs.org/text.asp?2017/10/2/196/207260


  Introduction Top


Prosthodontic rehabilitation of edentulous patients is a difficult task because the loss of all teeth leads to many problems in edentulous patients such as maintenance of health, poor mastication, poor esthetics, improper speech, nutrition, and social interactions. To overcome this problem, the most common treatment modality followed is complete denture fabrication, but due to the material properties of denture base material and the inability to maintain the denture hygiene by elderly patients because of their decreased manual abilities, poor dexterity, and various systemic diseases, maintaining oral and denture hygiene is difficult.

The formation of plaque on the surfaces of dentures among denture wearers often leads to halitosis, denture stomatitis, and chronic candidiasis. Denture plaque formation is facilitated by the presence of debris due to poor hygiene, irregularities in the acrylic resin. The best approach for controlling these deposits and infections is by implementing preventive measures such as maintenance of good oral hygiene and mechanical and chemical cleaning procedures for maintaining the denture.[1]

Various methods have been proposed to maintain denture hygiene. These methods are divided into mechanical methods, chemical methods, and microwave irradiation. Mechanical methods include brushing, sonic vibrators, and ultrasonic cleaner.[2] Denture cleansers are available in different forms such as tablets, pastes, and powders with a variety of manufacturer's claims for their relative efficacies.

Although most patients clean their dentures using manual brushing, this method when used solely has been considered one of the least efficient methods for biofilm control. The brushing method requires manual dexterity and visual acuity which are usually compromised in the elderly. Consequently, for biofilm control, daily immersion in cleaning solutions has been suggested to complement denture hygiene, and this combination has demonstrated greater results when compared to brushing alone.[3]

Several studies have identified Streptococci as a major constituent of the microflora of denture and Candida albicans as a major etiological significance in denture-induced stomatitis.

Diabetes mellitus is a common and increasing global health problem which causes several complications. Diabetics have an increased predisposition to the manifestations of oral diseases such as candidiasis, which is associated with poor glycemic control and therapeutic dentures. Wearing complete denture is also known as another risk factor, which can promote colonization of Candida, produce Candidal biofilm, and result in oral candidiasis. Association of denture and diabetes increases the incidence of oral candidiasis in patients with diabetes.[4]

Studies have reported on the comparison of chemical and mechanical denture-cleansing methods, but there is obvious lack of the studies comparing their effect on diabetic and nondiabetic denture biofilm. Hence, the present study was conducted to evaluate the effect of tablet and paste form denture cleansers on bacterial and Candidal count of diabetic and nondiabetic denture biofilm.


  Methodology Top


Forty individuals were included in the study (Group A: 20 diabetics and Group B: 20 nondiabetics). Both these two groups were further divided into 4 subgroups: Group 1A - Patients with diabetes who were given denture cleanser in tablet form (n = 10); Group 1B - Patients with diabetes who were given denture cleanser in paste form (n = 10); Group 2A: Nondiabetic individuals who were given denture cleanser in tablet form (n = 10); and Group 2B: Nondiabetic individuals who were given denture cleanser in paste form (n = 10).

The denture wearer patients selected were using the dentures for at least 6 months. All dentures were fabricated of heat-cured acrylic resin. Medical history was recorded for the patients. The diabetic status of the patients was determined by the history of previous diagnosis of diabetes, and the blood glucose levels were also determined using colorimetric method before the sample collection. The patients were assessed as nondiabetics and diabetics on the ground of fasting plasma glucose levels (normal = <110 mg/dl).

The experimental period was 21 days. All individuals were instructed to follow the following denture-cleansing guidelines.

  1. Denture brushing three times a day, after each meal (breakfast, lunch, and dinner) for 2 min, using STIM denture brush and tap water
  2. Rinsing the oral cavity with running water
  3. Keeping the dentures immersed in water while sleeping.


Experimental Groups 1 and 2, i.e., tablet form group, were instructed to use Clanden denture-cleansing tablets for 21 days by soaking the dentures in a container with water and one effervescent tablet (Clanden) for 5 min, after dinner along with the above instructions.

Experimental Groups 3 and 4, i.e., paste form group, were instructed to use Clanden denture-cleansing paste for 21 days by putting two pea-shaped drops on denture surface and brushing with STIM denture brush for 2 min along with above instructions [Figure 1].
Figure 1: Clanden tablets and paste along with STIM denture brush prescribed to the patient

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Samples were collected twice for each participant, before and after the use of denture cleanser for 21-day period, i.e., at baseline and after the experiment.

Microbiological evaluation

For microbiological sampling, maxillary and mandibular complete dentures were removed from patient's mouth and placed in a sterile container. Dentures were rinsed with distilled water to remove the saliva and then were placed on a sterile  Petri dish More Details containing 10 ml brain–heart infusion broth. Then, denture surfaces were brushed with a denture brush. This procedure was carried out in an inoculation hood to prevent sample contamination [Figure 2].
Figure 2: Sample collection by brushing the denture surface

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The resultant suspension (10 ml) was then transferred to a test tube and centrifuged for 4 min at a speed of 3000/rpm. Supernatant 9 ml liquid was decanted, and the remaining 1 ml liquid was used for culturing. This 1 ml liquid was mixed thoroughly in a vortex mixer, and 0.1 ml was taken from this liquid with a sterile 4 mm diameter inoculating loop and cultured on blood agar and chrome agar by lawn culture method [Figure 3] and [Figure 4]. The blood and chromogenic agar plates were prepared according to the manufacturer's instructions. Blood agar plates were incubated at 37°C for 24 h and chrome agar in biological oxygen demand at 29°C for 24 h.
Figure 3: Lawn culture on blood agar with sterile loop

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Figure 4: Lawn culture on CHROMagar with sterile loop

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The number of colonies of Streptococcus species on blood agar plate and number of colonies of Candida species on chrome agar were counted after 24 h of incubation, and the number of colony-forming unit (CFU)/ml was derived from the formula [Figure 5] and [Figure 6]. This process and colony counting was carried out by the same operator in laboratory to exclude the examiners bias. The resultant data were analyzed statistically to draw conclusions from the experimental data.
Figure 5: Bacterial colonization seen on blood agar after 24 h incubation

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Figure 6: Candidal colonization seen on CHROMagar after 24 h incubation period

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  Results Top


The collected data were tabulated and analyzed statistically using SPSS software version 20. To collectively compare the means of the study groups, two-way ANOVA tests were used (P < 0.05), pairwise comparison of the test group was performed using Tukey's multiple post hoc (P < 0.05), and dependent t-test was used for comparison of CFU counts in before and after treatment in diabetic and nondiabetic groups for both Streptococcus and Candida species.

The mean and standard deviation of baseline and after treatment log (CFU+1) counts were calculated for specimens (n = 10) in two groups (diabetic and nondiabetic) [Table 1] and [Table 2] and two forms (tablets and pastes) [Table 3] and [Table 4]. The mean values of log (CFU+1) counts of after treatment were 1.10 and 1.47, which were significantly less when compared to baseline values of 2.72 and 2.23 of Streptococcus species for tablet and paste form in nondiabetic patients, respectively. The similar values were found with the diabetic patients; the mean values of log (CFU+1) counts of after treatment were 0.63 and 1.57, which were significantly less when compared to baseline values of 2.79 and 2.24.
Table 1: Comparison of baseline and after treatment log (colony-forming units+1) counts of Candida in diabetic and nondiabetic groups by paired t-test

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Table 2: Comparison of baseline and after treatment log (colony-forming units+1) counts of Candida in tablet and paste forms by paired t-test

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Table 3: Comparison of baseline and after treatment log (colony-forming units+1) counts of Streptococcus in diabetic and nondiabetic groups by paired t-test

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Table 4: Comparison of baseline and after treatment log (colony-forming units+1) counts of Streptococcus in tablet and paste forms by paired t-test

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The mean values of log (CFU+1) counts of after treatment were 1.51 and 1.70, which were significantly less when compared to baseline values of 2.72 and 2.23 of Candida species for tablet and paste form in nondiabetic patients, respectively. The similar values were found with the diabetic patients; the mean values of log (CFU+1) counts of after treatment were 1.02 and 1.57, which were significantly less when compared to baseline values of 1.94 and 2.16.


  Discussion Top


For maintaining the health of oral mucosa in denture wearers, proper hygiene of removable denture is important. Dentures can harbor both oral bacterial and fungal microorganisms such as Streptococcus and Candida spp.[5]

According to a study done on maintenance of denture cleanliness by Marchini et al., only 26.3% of the patients removed their complete dentures overnight. They reported in their study that only 22.5% of the patients had been given instructions about denture cleansing, while the majority of the patients (77.5%) declared that they had not been given such instructions. They also reported that mechanical cleaning is the most prevalent method of hygiene maintenance among denture wearers.[6]

The study included forty old denture-wearing diabetic and nondiabetic individuals who were being prescribed either a denture cleanser tablet or a denture cleanser paste, to evaluate the effect of these denture cleansers on the oral microbial colonization. Old denture wearers of a minimum of 6 months were included in the study. The evaluation and comparison was carried out by counting the CFUs of Streptococcus and Candida on blood agar and chrome agar, respectively, after a period of 21 days of denture cleanser usage by these denture wearers. Patients on corticosteroid therapy and antifungal therapy, patients suffering from diseases compromising the immune system, and patients with existing clinical lesion of oral candidiasis and with the history of tobacco chewing and smoking which are known to predispose the patients to oral infections, were excluded from the study as they may alter the results of the study.

Results of the present study have shown that the usage of denture cleansers reduces the microbial count in both diabetic and noniabetic denture biofilms [Figure 7] and [Figure 8]. The uses of effervescent tablets were significantly more effective against both bacteria and fungi than abrasive paste. Results of the present study support the observations of other studies.[5],[7],[8] This observation can be explained by the mechanism of action of these denture cleansers.
Figure 7: Candidal colonization on CHROMagar after 21 days of denture cleanser usage

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Figure 8: Bacterial colonization on Blood agar after 21 days of denture cleanser usage

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Effervescent denture-cleansing tablets contain alkaline peroxides; the one used in the present study contains sodium perborate monohydrate. These peroxides produce an effervescent alkaline solution of hydrogen peroxide, containing active oxygen, when in contact with water. The effervescence exerts a mechanical action by removing debris, and the oxygen has antimicrobial and stain-removing effects.[9]

On the other hand, a denture-cleansing paste typically contains abrasive particles to aid in mechanical cleaning of teeth; the one used in the present study contains hydrated silica as an abrasive. These abrasives abrade the denture surface and create roughness and may result in increased adherence of microorganisms.[5] The paste used in the present study also has triclosan as one of its ingredients which is an antimicrobial agent. This might be the reason for the reduction in the microbial count, but it was not as effective as the effervescent tablets. In the present study, there is a significant reduction in bacterial count after the use of abrasive paste, but the reduction in Candidal counts is not significant.

Denture-cleansing tablets showed a significant reduction not only in Streptococcus but also in Candida counts in both diabetic and nondiabetic denture biofilms. The results were similar to the study done by Nikawa et al.[10] and de Freitas Fernandes et al.[11]

On the contrary, many authors have reported in their studies that effervescent tablets are effective against bacterial count but not against Candidal count. They have suggested that, in a multispecies biofilm, bacteria such as Streptococci may be placed in more superficial layers than Candida, which protect these yeast cells from denture cleansers. Drake et al. suggested that Streptococcus mutans with its capacity of secreting polysaccharides produce a protective barrier of glucan that limits the exposure of yeast cells to denture cleansers' antimicrobial activity.[12]

In the present study, patients were instructed to brush their dentures three times a day along with the use of denture cleansers. That might explain the reduction in Candidal count as the glucan barrier may have been cleaned by brushing.

Denture wearers in general, whether they are diabetic or nondiabetic, are at greater risk of oral candidiasis. In the present study, there is a slight increase in Candidal count in diabetics as compared to the nondiabetics though the difference is statistically insignificant. This observation is consistent with the study done by Daniluk et al.[13] However, many studies have reported a statistically significant increase in Candidal count of diabetic individuals when compared to nondiabetic individuals.[4],[14]

Most of the reported studies have isolated Candida species as a whole; however, in the present study, different Candida species were also identified based on the color displayed by them on CHROMagar [Figure 9] and [Figure 10]. In the present study as expected Candida albicans was the most common isolate, frequency of nonalbicans Candida was more common in diabetic denture biofilm as compared to nondiabetic denture biofilm, which is inconsistent with the reports of other studies.[10],[15]Candida tropicalis and Candida krusei were isolated only from diabetics, and Candida glabrata was isolated from nondiabetics. Similar results were observed in the study done by Lotfi-Kamran.[4]
Figure 9: Different species of Candida based on color displayed by them on CHROMagar

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Figure 10: Different species of Candida based on color displayed by them on CHROMagar

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After the results were analyzed by applying various statistical tests, as mentioned earlier, it was concluded that there was a significant difference between the CFU counts of diabetic and nondiabetic denture biofilm after 21 days of usage of denture cleansers. However, effervescent tablets have shown more antibacterial and antifungal effects as compared to the paste form. The effervescent tablets were more effective against Candida than the paste form. However, a statistically insignificant difference was found between the microbial colonization in diabetic and nondiabetic individuals. Thus, the data support the rejection of the null hypothesis. It is now accepted that if the denture cleansers are used properly along with other denture-maintaining guidelines and the patient maintains a good oral hygiene, they can be an asset to a dentist's armamentarium and also in maintaining patients' oral and denture hygiene.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Chethan MD, Azhagarasan NS, Miglani S, Mohammed HS, Hari Prasad A. Microbiological evaluation of the effectiveness of commercially available denture cleansing agents. Int J Drug Dev Res 2011;3:159-72.  Back to cited text no. 1
    
2.
Budtz-Jørgensen E. Materials and methods for cleaning dentures. J Prosthet Dent 1979;42:619-23.  Back to cited text no. 2
    
3.
Neppelenbroek KH. The importance of daily removal of the denture biofilm for oral and systemic diseases prevention. J Appl Oral Sci 2015;23:547-8.  Back to cited text no. 3
    
4.
Lotfi-Kamran MH, Jafari AA, Falah-Tafti A, Tavakoli E, Falahzadeh MH. Candida colonization on the denture of diabetic and non-diabetic patients. Dent Res J (Isfahan) 2009;6:23-7.  Back to cited text no. 4
    
5.
Kiesow A, Sarembe S, Pizzey RL, Axe AS, Bradshaw DJ. Material compatibility and antimicrobial activity of consumer products commonly used to clean dentures. J Prosthet Dent 2016;115:189-98.e8.  Back to cited text no. 5
    
6.
Marchini L, Tamashiro E, Nascimento DF, Cunha VP. Self-reported denture hygiene of a sample of edentulous attendees at a university dental clinic and the relationship to the condition of the oral tissues. Gerodontology 2004;21:226-8.  Back to cited text no. 6
    
7.
Dills SS, Olshan AM, Goldner S, Brogdon C. Comparison of the antimicrobial capability of an abrasive paste and chemical-soak denture cleaners. J Prosthet Dent 1988;60:467-70.  Back to cited text no. 7
    
8.
Yadav R, Yadav VS, Garg S, Mittal S, Garg R. Effectiveness of different denture cleansing methods on removal of biofilms formed in vivo. J Cranio Maxillary Dis 2013;2:22-7.  Back to cited text no. 8
    
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Lucena-Ferreira SC, Ricomini-Filho AP, Silva WJ, Cury JA, Cury AA. Influence of daily immersion in denture cleanser on multispecies biofilm. Clin Oral Investig 2014;18:2179-85.  Back to cited text no. 9
    
10.
Nikawa H, Yamamoto T, Hamada T, Sadamori S, Agrawal S. Cleansing efficacy of commercial denture cleansers: Ability to reduce Candida albicans biofilm activity. Int J Prosthodont 1995;8:527-34.  Back to cited text no. 10
    
11.
de Freitas Fernandes FS, Pereira-Cenci T, da Silva WJ, Filho AP, Straioto FG, Del Bel Cury AA. Efficacy of denture cleansers on Candida spp. biofilm formed on polyamide and polymethyl methacrylate resins. J Prosthet Dent 2011;105:51-8.  Back to cited text no. 11
    
12.
Drake D, Wells J, Ettinger R. Efficacy of denture cleansing agents in an in vitro bacteria-yeast colonization model. Int J Prosthodont 1992;5:214-20.  Back to cited text no. 12
    
13.
Daniluk T, Tokajuk G, Stokowska W, Fiedoruk K, Sciepuk M, Zaremba ML, et al. Occurrence rate of oral Candida albicans in denture wearer patients. Adv Med Sci 2006;51 Suppl 1:77-80.  Back to cited text no. 13
    
14.
Bartholomew GA, Rodu B, Bell DS. Oral candidiasis in patients with diabetes mellitus: A thorough analysis. Diabetes Care 1987;10:607-12.  Back to cited text no. 14
    
15.
Abu-Elteen KH, Hamad MA, Salah SA. Prevalence of oral Candida infections in diabetic patients. Bahrain Med Bull 2006;28:1-8.  Back to cited text no. 15
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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