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| ORIGINAL ARTICLE |
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| Year : 2016 | Volume
: 5
| Issue : 2 | Page : 63-69 |
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“A comparative evaluation of the effects of different commercially available denture adhesives on the growth of Candida species in diabetic and nondiabetic subjects:” An In vivo Study
Abhishek Borole, KT Roopa, Priti Vasant Khandelwal
Department of Prosthodontics, College of Dental Sciences, Davangere, Karnataka, India
| Date of Web Publication | 25-Oct-2016 |
Correspondence Address:
Priti Vasant Khandelwal
Flat No. 303, Plot No. 123, Balaji Heights, Sector 50 (New), Nerul West, Navi Mumbai - 400 706, Maharashtra
India
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/2277-4696.192973
Purpose: Denture patients should use denture adhesives cautiously. Difficulty in maintaining good oral hygiene can be caused by the presence of a thick deposit of adhesive which may act as a reservoir of Candida albicans. In diabetic condition, chances of infections are more. Therefore, this study was carried out to investigate the effects of different commercially available denture adhesives on the growth of Candida species in diabetic and nondiabetic patients. Materials and Methods: Sixty edentulous subjects were divided into two main groups and six subgroups based on diabetes and use of three denture adhesives. The swab sample and nonstimulated saliva were collected at the time of denture insertion and 14 days after use of the adhesives. Microbiological procedures were carried out following the same. The data collected in terms of colony forming unit and colony forming unit per milliliter (CFU and CFU/ml) was subjected to statistical analysis. Results: Intra- and inter-group comparison showed no significant difference to exist for both test and control group (CG); although, there was a minimal increase in the number of CFU/ml of Candida species at 14 days. Interpretation and Conclusion: There was an overall increase in the number of CFU/ml of Candida species following the use of denture adhesives in CG and test groups and after 14 days of use of denture adhesive. The mean percentage increase in CFU/ml was not of any clinical significance. Keywords: Colony forming unit counts, denture adhesive, diabetic, serial dilutions
How to cite this article:
Borole A, Roopa K T, Khandelwal PV. “A comparative evaluation of the effects of different commercially available denture adhesives on the growth of Candida species in diabetic and nondiabetic subjects:” An In vivo Study. J Dent Allied Sci 2016;5:63-9 |
How to cite this URL:
Borole A, Roopa K T, Khandelwal PV. “A comparative evaluation of the effects of different commercially available denture adhesives on the growth of Candida species in diabetic and nondiabetic subjects:” An In vivo Study. J Dent Allied Sci [serial online] 2016 [cited 2023 Jun 5];5:63-9. Available from: https://www.jdas.in/text.asp?2016/5/2/63/192973 |
| Introduction | |  |
The clinical procedures during fabrication of complete denture impression making are aimed to obtain an effective border seal.[1] When there is a necessity for optimal denture retention patients frequently resort to the use of denture adhesives; which supposed to increase retention and stability.[2],[3]
The presence of a thick deposit of adhesive may act as a reservoir of microbial biofilms like Candida albicans which has deleterious effects on the oral mucosa.[4],[5] Patients with diabetes mellitus has reduced salivary flow and low salivary buffering capacity. Thus, the misuse of denture adhesives presents a higher susceptibility to infections in these patients.[6],[7],[8]
The purpose of this pilot study was to investigate the effects of different commercially available denture adhesives on the growth of Candida species in diabetic and nondiabetic patients.
| Materials and Methods | | |
Subject selection and protocol
Null hypothesis
There is no difference in C. albicans counts in diabetic and nondiabetic patients on using commercially available denture adhesives.
From patients attending the Department of Prosthodontics, College of Dental Sciences, Davangere, Karnataka, India for replacement of lost teeth with complete dentures, sixty completely edentulous patients of both genders (40–70 years) were selected. The diabetic status of the subject was evaluated in consultation with a diabetologist per guidelines laid down by the American Diabetes Association. The subjects were divided into two main groups, namely, control group (CG) (nondiabetic) and test group (TG) (diabetic).
Three denture adhesives were used for control and TG with 10 individuals in each type of denture adhesive. Instructions were given to patients for cleaning of the denture with a soft dental brush in the morning and after meal on occlusal and polished surfaces and cleaning with fingers only under tap water for intaglio surface. The patient was also asked to apply a new layer of denture adhesive every day.
- A1: Fixon, Super Grip Fixon Cream, ICPA Health Products Ltd., Ankleshwar, Gujarat, India
- A2: Fittydent, Fittydent International GMBH, Pinkafeld, Austria
- A3: Denofit, Dent Aids, Goran Pharma Pvt. Ltd., Sihor, Gujarat, India.
All these individuals were subjected to evaluate candidial growth before denture insertion (at baseline) and at 14 days since the start of use of prescribed denture adhesive, which was prescribed at the time of denture insertion.
Selection criteria
- CG: Edentulous subjects without any significant medical history were selected.
- TG: Known controlled diabetic completely edentulous subjects without any other significant medical history
- CG and TG had no history of antibiotic therapy for at least 6 weeks before participation in the study.
After knowing the diabetic status of the completely edentulous subjects, thirty controlled diabetic (TG) and thirty subjects with no significant medical history (CG) were selected after obtaining informed consent.
Collection of samples
Samples were collected by two methods:
- Sterile cotton swab was used to obtain first sample from palatal mucosa at baseline and from intaglio surface of denture at 14th day [Figure 1] and [Figure 2][9]
- Subjects were asked to deposit nonstimulated saliva into a sterile container over a duration of 5 min [Figure 3].[10]
 | Figure 2: Swab sample from palatal intaglio surface of maxillary complete denture
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Samples were collected at baseline and after 14 days for each denture adhesive. Three serial dilutions were made for each swab and each sample of saliva collected from subjects to prepare sample solutions.
Culture plates containing sterile culture medium were created with Sabouraud's Dextrose Agar with chloramphenicol (Medium 3. HiMedia Laboratories Pvt. Ltd., Mumbai, India).[7]
Inoculation of culture plates
From each of the serial diluted test tube 0.05 ml solution was inoculated into subsequent culture plates using a spread plate method and kept into an incubator for 37°C for 48 h in aerobic condition.[11]
After the incubation period, the macroscopically (spherical shape, white, matte with a porcelain appearance, diameter between 1 and 8 mm) and microscopically (Gram staining) the colonies of Candida species were identified [Figure 4] and [Figure 5].
Then, culture plates for both swab and saliva sample were counted by using colony counting device (Medox Bio Colony Counter, Medox Biotech Pvt. Ltd., Chennai, India).
Subsequently, the same microbiological procedure was carried out for second samples. The data thus collected regarding CFU/ml was subjected to statistical analysis.
Data analysis
Data collected by experiments were computerized and analyzed using the Statistical Package for Social Sciences (SPSS) version 17.0 (IBM Corporation, Armonk, New York, US).
Mean % increase, nonparametric tests, namely, Wilcoxon signed-rank test for intragroup comparing microbial parameter (CFU/ml) with baseline and at 14 days and Mann–Whitney U-test for group-wise comparison between control and TGs; were used for testing the statistical significance.
For all tests, a P value of 0.05 or less was considered for statistical significance.
| Results | | |
The purpose of this clinical laboratory study was to evaluate and compare the effects of different commercially available denture adhesives on the growth of Candida species in diabetic and nondiabetic subjects.
There was an overall increase in the number of CFU/ml for all three denture adhesive in the CG at 14 days. The mean percentage increase ranging from 0.99% (A3) to 3.16% (A1) for swab sample and 0.094% (A1) to 1.44% (A3) for saliva sample in CG and 2.43% (A1) to 3.61% (A2) for swab sample and 1.21% (A3) to 2.90% (A2) for saliva sample for TG [Table 1], [Table 2] and [Graph 1]. | Table 1: Comparison of mean percentage increase in CFU/ml values of swab and saliva samples of control group between adhesives A1, A2 and A3 at 14 days
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 | Table 2: Comparison of mean percentage increase in CFU/ml values of swab and saliva samples of test group between adhesives A1, A2 and A3 at 14 days, and P value
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In CG for comparison of number of CFU/ml in swab sample at baseline and at 14 days, mean CFU/ml value at baseline ranged from 77.67 (standard deviation [SD] =5.12) to 80.33 (SD = 8.23) and for comparison of number of CFU/ml in saliva sample at baseline and at 14 days; the mean CFU/ml value at baseline ranged from 111.43 (SD = 10.30) to 119.67 (SD = 12.16). There was a slight increase in the CFU/ml value of the sample at 14 days when compared to baseline values in both samples. Comparison of the baseline values with that at 14 days was done using Wilcoxon signed-rank test (nonparametric). No significant differences were observed in CFU/ml at 14 days; as for swab sample - A1: P = 0.465, A2: P = 0.705, A3: P = 0.066, for saliva sample - A1: P = 0.104, A2: P = 0.892, A3: P = 0.655 [Table 1] and [Graph 1], [Graph 2].
In TG a comparison of number of CFU/ml in swab sample at baseline the mean CFU/ml value at baseline ranged from 79.25 (SD = 6.75) to 82.5 (SD = 9.24) and at 14 days test there was a slight increase in the CFU/ml value of the sample at 14 days (A1 = 82.29, A2 = 82.22, A3 = 84.75) when compared to baseline values. In saliva sample, the mean CFU/ml value at baseline ranged from 113.11 (SD = 10.05) to 122.25 (SD = 15.69). There was a slight increase in the CFU/ml value of the sample at 14 days (A1 = 121.11, A2 = 116.5, A3 = 123.75) when compared to baseline values. Comparison of the baseline values with that at 14 days was done using Wilcoxon signed-rank test (nonparametric). No significant differences were observed in CFU/ml at 14 days for both the samples for swab sample - A1: P = 0.066, A2: P = 0.671, A3: P = 0.260, for saliva samples - A1: P = 0.267, A2: P = 0.396, A3: P = 0.074 [Table 2] and [Graph 1],[Graph 2].
A comparison of swab and saliva sample CFU/ml between control and TGs using three different denture adhesives at baseline and at 14 days showed no significant differences [Table 1] and [Table 2].
Comparison of swab and saliva sample CFU/ml between control and TGs at baseline using Mann–Whitney U-test (nonparametric). No significant differences were observed between the groups using three different denture adhesives. For swab sample - A1: P = 0.710, A2: P = 0.573, A3: P = 0.414, for saliva samples - A1: P = 0.694, A2: P = 0.758, A3: P = 0.755 [Table 3] and [Graph 1], [Graph 2]. | Table 3: Comparison of swab sample CFU/ml between control and test group at baseline and at 14 days and comparison of saliva sample CFU/ml between control and test group at baseline and at 14 days
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Comparison of swab sample CFU/ml between control and TGs at 14 days using Mann–Whitney U-test (nonparametric). No significant differences were observed between the groups using 3 different denture adhesives, for swab samples - A1: P = 0.694, A2: P = 0.470, A3: P = 0.463, for saliva samples - A1: P = 0.470, A2: P = 0.574, A3: P = 0.694 [Table 3].
| Discussion | | |
Satisfactory retention is a prerequisite for the patient's acceptance of complete denture prosthesis.[8] The retention of the denture in the oral cavity is affected by a combination of factors such as adhesion, cohesion, atmospheric pressure, interfacial surface tension, and viscosity of the saliva. A deficiency of any one of these factors may compromise the retention of the prosthesis, thereby adversely affecting the patient's acceptance of prosthesis.
Although there is no substitute for application of good clinical techniques to maximize the retention and in addition, the support and stability of complete dentures, a clinician is often faced with demanding situations that are complicated by factors such as the patient's oral anatomy, systemic health and cost of treatment, where other alternatives must be found.
The use of denture adhesives began about the same time as modern dentistry in the late 18th century.[8] The denture adhesives are indicated for use in situ ations that require extra security of stable dentures, patients with systemic diseases, in patients undergoing extensive maxillofacial surgery to retain the large prosthesis and patients undergoing drug therapy resulting in xerostomia or similar situations.[2]
Among systemic conditions, diabetes mellitus is one of the most prevalent diseases worldwide and is commonly found in dental patients. Oral manifestations have been reported such as vascular alterations, blood coagulation, tissue regeneration times above normal, residual bone resorption, candidiasis, and loss of resiliency of the oral mucosa which is necessary for the good adaptation of complete dentures.[6] Sennerby et al. revealed that patients with diabetes present a lower level of retention of complete dentures than patients without the disease, as diabetes increases the osteoclastic activity of bone tissue in the mandible and maxilla. It is possible that diabetic denture wearers might require the use of denture adhesives to improve the retentive ability of the prosthesis over a period.
The denture adhesives are marketed as powder, cream, paste, and strip form that may be used by the patient in a prescribed manner. Regarding composition all three adhesives (Fixon, Fittydent, Denofit) which were used in this study are similar with the principal ingredients being.
- Fixon (A1) (ICPA) - white petrolatum, aerosil (fumed colloidal silica), light petrolatum, gantrez, cellulose gum, coloring, and flavoring agents
- Fittydent (A2) (Fittydent International GMBH) - sodium carboxymethylcellulose, solution of polyvinylacetate, petrolatum
- Denofit (A3) (Dent Aids) - gantrez (bio-adhesive), cellulose gum, white petrolatum, mineral oil.
The most popular and successful products consist of mixtures of salts of; short acting polymer (carboxymethyl cellulose), antimicrobial agents and long-acting polymer (polyvinylmethyl ether maleate or gangtrez).
In general, it is agreed that the denture adhesives act by providing an interface between the denture base materials and the oral mucosa and an intermediary of a thin film of saliva. Denture adhesives optimize interfacial forces and enhance retention by increasing the adhesive and cohesive properties and viscosity of the medium lying between the denture and its basal seat and eliminating voids in between.
Furthermore, the molecules of the adhesive establish a chemical bond through the carboxylic groups with the surface proteins of the mucosa cells.
The difficulties that arise in prosthetically rehabilitating the aged may not only be attributed to denture construction but also to associated conditions observed frequently such as denture stomatitis secondary to candidal infection.
Candida species are found in the oral cavity in 25–50% of healthy individuals. When only patients with complete dentures are taken into consideration, the values increase from 60% to 100%.[10] Among the edentulous elderly individuals, this pathogen has been observed in the oral cavity in the region of the buccal mucosa and the vault of the palate. In denture wearers, candidiasis is developed by the adhesion of C. albicans to the tissue surface of the maxillary denture base, which serves as an effective reservoir of microorganisms.[5]
C. albicans has been demonstrated on the fitting surface of the dentures. This was attributed to the acidic pH prevalent under the fitting surface of the dentures which aided the proliferation of the fungi. Olsen in 1974 considered the fitting surface of the denture as a reservoir of infection.[9] Weinstein has reported that the yeast, C albicans was isolated in greater quantities from the saliva of patients with diagnosed diabetes mellitus than from patients without a diagnosis of diabetes.[8]
Little is known about the influence of denture adhesives on the oral microflora, particularly in patients with a predisposition to prosthetic stomatitis such as in diabetic patients, in which the microbial components might be involved.[10]
Based on this information, the aim of this study was to evaluate the effects of different commercially available denture adhesives on the growth of Candida species in diabetic and nondiabetic subjects for 14 days.
In this study, the candidial growth measured directly in terms of absolute number of colony forming units (CFUs) and colony forming units per milliliter of sample (CFUs/ml) following plating of serial. To ensure that colony counts will be within the range, the original inoculums (saliva and swab samples) are diluted several times in a process called serial dilution.
Plate counts measure the number of viable cells after 24 h. A colony often results, from short segments of a chain or from a microbial clump. To reflect this plate counts often reported as CFU.
There was an overall increase in the number of CFU/ml for all the three denture adhesives in the CG at 14 days. The mean percentage increase ranged for samples were however too minimal to merit any clinical significance.
These findings indicate that regardless of the diabetic status of a subject the choice of denture adhesive may not be a causative factor in the development of oral candidiasis, up to the 14 days period.
Since all the adhesives used in this study were of similar composition, a minimal variation was observed between the different denture adhesives used regarding CFU/ml at 14 days.
On comparison of a number of CFU/ml of nondiabetic (CG) and diabetic (TG) subjects at 14 days showed no significant differences were seen for any of the adhesives used regardless of the method of sample collection, i.e., by swab.
These findings may be explained based on observations by Soysa et al. who discussed the relationship between diabetes and infection. They suggested that controversy existed regarding whether poor glycemic control predisposes to oral candidial infection in diabetic subjects.[7]
It may be noted that both groups were compared at baseline regarding number CFU/ml of Candida species as evidenced by the lack of any significant differences between the groups. Although TG did show a marginal increase in the number of CFU/ml at 14 days, a similar difference was observed in the calculated value for nondiabetic subjects (CG) at baseline and at 14 days.
A study conducted by Lima et al. stated that patients with a diagnosis of diabetes mellitus present a higher susceptibility to infections due to a deficiency in polymorphonuclear leukocytes. An increased risk of infections has been observed with reduced salivary flow, leading to low salivary buffering.[6]
It can be seen from this study that denture adhesives did not show any significant increase in the growth of Candida species in both nondiabetic and diabetic subjects. Other factors such as poor oral hygiene, poor systemic health, or immunocompromised state may be responsible for the proliferation of Candida species and must be further investigated.
Some limitations of this study were the absence of multiple brands of denture adhesives, absence of different forms of denture adhesives, small study population and relatively short trial period, patients varying cleaning practices for dentures; although, instructions for cleaning dentures were given in same way to all patients. Further studies should include longitudinal assessment of Candida species proliferation over an extended period.
| Conclusion | | |
Within limitations of this study and after analyzing statistical data following conclusions can be drawn. There was an overall increase in the numbers of CFU/ml of Candida species following the use of denture adhesives (A1: Fixon, A2: Fittydent, A3: Denofit) in both CG (nondiabetic) and TGs (diabetic) after 14 days, following conclusions are made-
- Increase in CFU/ml of Candida species in CG, for swab sample is A1 > A2 > A3; for saliva sample is A3 > A2 > A1
- Increase in CFU/ml of Candida species in TG, for swab sample is A2 > A3 > A1; for saliva sample is A2 > A1 > A3
- There was an increase in candidial CFU/ml values for the controlled diabetic subjects (TG) as compared to that of the nondiabetic subjects (CG).
However, the mean percentage increase in CFU/ml was not of any clinical significance in both the groups and both samples for three denture adhesives.
Declaration of patient consent
The authors certify that they have obtained all appropriate patient consent forms. In the form the patient(s) has/have given his/her/their consent for his/her/their images and other clinical information to be reported in the journal. The patients understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed.
Financial support and sponsorship
Equipment - Department of Prosthodontics, College of Dental Sciences, Davangere.
Conflict of interest
There are no conflicts of interest.
| References | | |
| 1. | Ow RK, Bearn EM. A method of studying the effect of adhesives on denture retention. J Prosthet Dent 1983;50:332-7.  |
| 2. | Panagiotouni E, Pissiotis A, Kapari D, Kaloyannides A. Retentive ability of various denture adhesive materials: An in vitro study. J Prosthet Dent 1995;73:578-85. |
| 3. | Grasso JE. Denture adhesives. Dent Clin North Am 2004;48:721-33. |
| 4. | Makihira S, Nikawa H, Satonobu SV, Jin C, Hamada T. Growth of Candida species on commercial denture adhesives in vitro. Int J Prosthodont 2001;14:48-52. |
| 5. | Ramage G, Tomsett K, Wickes BL, López-Ribot JL, Redding SW. Denture stomatitis: A role for Candida biofilms. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2004;98:53-9. |
| 6. | Cristina de Lima D, Nakata GC, Balducci I, Almeida JD. Oral manifestations of diabetes mellitus in complete denture wearers. J Prosthet Dent 2008;99:60-5. |
| 7. | Soysa NS, Samaranayake LP, Ellepola AN. Diabetes mellitus as a contributory factor in oral candidosis. Diabet Med 2006;23:455-9. |
| 8. | Peters RB, Bahn AN, Barens G. Candida albicans in the oral cavities of diabetics. J Dent Res 1966;45:771-7. |
| 9. | Olsen I. Denture stomatitis. Occurrence and distribution of fungi. Acta Odontol Scand 1974;32:329-33. |
| 10. | Oliveira MC, Oliveira VM, Vieira AC, Rambob I. In vivo assessment of the effect of an adhesive for complete dentures on colonisation of Candida species. Gerodontology 2010;27:303-7. |
| 11. | Tortora GJ, Funke BR, Christine L. Microbiology: An Introduction. 3 rd ed. Redwood City, Calif: Benjamin/Cummings Pub. Co.; 2012. p. 171-7. |
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3]
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