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ORIGINAL RESEARCH  
Year : 2013  |  Volume : 24  |  Issue : 5  |  Page : 645
An in vitro study on effect of Delmopinol application on Candida albicans adherence on heat cured denture base acrylic resin: A thorough study


Department of Prosthodontics, Government College of Dentistry, Indore, Madhya Pradesh, India

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Date of Submission03-Jan-2013
Date of Decision22-Mar-2013
Date of Acceptance13-May-2013
Date of Web Publication21-Dec-2013
 

   Abstract 

Background: Denture-related stomatitis is probably the most common form of oral candidiasis and its reported prevalence varies widely ranging up to 65%. In this thorough study, we evaluate the effect of Delmopinol application on Candida albicans adherence on heat cured denture base acrylic resin.
Materials and Methods: A total of 40 specimens of heat-cured acrylic resin were made, 20 specimens were contaminated before Delmopinol treatment and 20 specimens were contaminated after Delmopinol treatment. The each specimen in each tube was individually transferred to a spectrophotometer at 530 nm wavelength in order to measure the turbidity degree, through the transmittance. Aliquots of 10 μL of each tube was then collected and inoculated into agar sabouraud plates containing 500 mMol/L of sucrose, which was incubated for 24 hours at 37°C, in order to check microbial growth.
Results: Mean of the turbidity Degree of contamination after Delmopinol application (1440.80, colony-forming unit [CFU] [×10 6 /ml]) was significantly higher than the mean of the turbidity degree of contamination before Delmopinol application (550.85 CFU [×10 6 /ml]).
Conclusion: Heat-cured acrylic resin shows greater reduction in adherence of Candida albicans by contamination after Delmopinol application as compared with contamination before Delmopinol application.

Keywords: Candida albicans , delmopinol, denture induced stomatitis, heat cure acrylic resin

How to cite this article:
Jain D, Shakya P. An in vitro study on effect of Delmopinol application on Candida albicans adherence on heat cured denture base acrylic resin: A thorough study. Indian J Dent Res 2013;24:645

How to cite this URL:
Jain D, Shakya P. An in vitro study on effect of Delmopinol application on Candida albicans adherence on heat cured denture base acrylic resin: A thorough study. Indian J Dent Res [serial online] 2013 [cited 2019 Dec 11];24:645. Available from: http://www.ijdr.in/text.asp?2013/24/5/645/123423
The synthetic acrylic resins have a long, clinically proven history of use for dentures since they exhibit adequate physical, mechanical and esthetic properties; [1] however, they are susceptible to microbial adhesion, leading to denture stomatitis. [2] Every denture causes at least some harm to the underlying tissues either directly or indirectly. One important aspect of this harm is denture related stomatitis. The presence of dentures may be the triggering local factor for diseased oral mucosa. [3] Denture related stomatitis is frequent among denture wearers and its reported prevalence varies widely ranging up to 65%. [4]

Denture-related stomatitis is an inflammatory process of the mucosa; generally, a denture with a multifactorial etiology. [5],[6] Plaque on the tissue surface of the denture is unquestionably a major etiologic factor in the pathogenesis of denture stomatitis, inflammatory papillary hyperplasia and chronic candidiasis. [7]

Denture plaque is defined as a dense microbial layer comprising micro-organisms and their metabolites and it is known to contain more than 10 11 organisms per gram in wet weight. Light and electron microscopic studies have revealed that denture plaque has essentially the same structure and the composition of microbial flora as dental plaque on natural teeth, except for an increased number of Candida species, which accompanies denture-induced stomatitis. [8]

There are several factors, which affect Candida albicans adhesion on acrylic resin surface; quality and quantity of saliva, surface properties of acrylic resin [9] and oral bacteria. [5]

The adherence of C. albicans to acrylic resin has been thought to be an essential pre-requisite for successful colonization; subsequent plaque formation and development of pathogenesis [10] that is correlates positively with their roughness. [11],[12] The surface properties of denture base materials are of paramount importance for maintaining the structural and functional integrity of the denture-covered oral mucosa.

Control of denture plaque is facilitated if the denture surfaces, including the tissue surface, are well-polished as it will prevent the accumulation of debris. [13] The tissue surfaces are usually left as they were when removed from the cast; thus, it presents irregularities and microscopic pores that facilitate bacterial and fungal colonization.

It seems that denture hygiene is often neglected by both patients and dentists. Denture cleanliness is reported to be generally poor and denture wearers seem to easily adjust to unclean dentures [14] and poor denture hygiene might dispose to denture stomatitis. [15]

In principle, mechanical cleansing is an effective means of improving denture cleanliness, but major disadvantage of it, is loss of surface details, which ultimately cause loss of denture retention. Chemical denture cleansers (Antiseptic and Disinfecting agents) might be an important additive to mechanical cleansing, especially among geriatric and/or handicapped patients. Chemical disinfectants also prevent cross contamination from the prosthesis. [16],[17]

The morpholinoethanol derivative Delmopinol is a third generation of chemical-plaque control agents. Delmopinol is a tertiary amine surfactant that is used to counteract dental plaque formation and dissolves newly formed plaque. The adsorption of Delmopinol is complex and strongly influenced by pH and concentration [18] and adsorbed on both hydrophobic and hydrophilic surface. [19]

Plaque studies involving Delmopinol revealed that the nascent bio-film was loosely adherent [20] and that there was a significant reduction in the proportion of Dextran-producing Streptococci. [21] There was no colonization by Candida or major shift in bacterial composition in the active group nor was there any decrease in susceptibility to Delmopinol. [22],[23]

There are very few studies reported in the literature showing the effect of Delmopinol on deposition of plaque on acrylic resin denture tissue surface. Therefore, it is pertaining to taken up an in-vitro study for evaluation and comparison of effect of Delmopinol application on heat-cured acrylic resin on C. albicans adherence.


   Materials and Methods Top


Preparation of acrylic resin specimens

Prefabricated acrylic resin sheet (Polytek Industries, Indore, Madhya Pradesh, India) was used to fabricate dies having 10 mm × 10 mm × 5 mm dimensions, were invested in flasks and mold was obtained. Thin layer of acrylic resin separating media film (Acralyn-H, Asian Acrylates, Mumbai, India) was applied and was packed with plastic stage heat cured acrylic resin (Acralyn-H, Asian Acrylates, Mumbai, India). The flasks were pressed slowly with a hydraulic pressure until the excess material extruded from the borders and then screwed. Polymerization procedure was performed; according to the cycle described by the manufacturer's instructions and specimens were cooled at room temperature for 30 min. and left in tap water for 15 min. Forty specimens of dimension 10 mm × 10 mm × 5 mm were fabricated with a heat-cured acrylic resin and were removed after opening the flasks. No finishing and polishing procedure was carried out in order to simulate the tissue surface of a complete denture. Only remaining excess was removed with the aid of 320-grit wet sandpaper.

Microbiological test

Before culture of C. albicans on acrylic resin plate, all specimens are sterilized by immersion in 2% Gluteraldehyde (PKS Phama Pvt. Ltd. Karnataka, India) for 30 min to prevent contamination and placed in a sterilized container. The specimens were immersed in distilled water (Hindustan Pharmaceuticals, Barauni) in order to promote the maximum water sorption to prevent, when in culture, the occurrence of distortion and the release of residual monomer after polymerization. Pure culture of C. albicans was grown on agar sabouraud plates (Himedia laboratory Pvt. Ltd, Mumbai) containing 500 mMol/L of sucrose at 25°C. After 24 hours, the colonies were suspended in tubes containing 5 mL of brain heart infusion (BHI) broth (Himedia laboratory Pvt. Ltd, Mumbai). The cell suspension in each tube was adjusted spectrophotometrically at 800 nm (O.D.800) to match the transmittance of 90 T (equivalent to 0.5 McFarland scale = 1.5 × 108 colony-forming unit [CFU]). The C. albicans (ACTT - 10231) strain was obtained from Department of Microbiology, Calcutta. Next, the specimens were placed into the tubes containing BHI plus inoculums and remained for 11 hours at 37°C in order to favor initial colonization of the acrylic resin surfaces and colony count was calculated at this stage. Each specimen was first washed with saline (Hindustan Pharmaceuticals, Barauni) after immersion in the contaminated culture broth. Saline excess was removed with a gentle compression of sterile gauze. Then, the disinfection step was performed by application of Delmopinol (Sinclair pharmaceuticals limited, Sweden, U.K. Marketed by Wockhardt limited In India) for 48 hours. Each specimen was then washed again with saline and the excess was removed with sterile gauze. It was then transferred to individual tubes containing 5 mL of BHI broth. After 24 hours of incubation, the tubes were individually transferred to a spectrophotometer at 530 nm wavelength in order to measure the turbidity degree [Table 1], through the transmittance. Aliquots of 10 μL of each tube was then collected and inoculated into agar sabouraud plates containing 500 mMol/L of sucrose, which was incubated for 24 hours at 37°C, in order to check microbial growth. The purity of the positive cultures was confirmed by gram staining, by colony morphology on agar plates.
Table 1: Value of CFU (×106/ml) obtained in the heat-cured acrylic resin specimens


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   Results Top


Turbidity degree is inversely proportional to the C. albicans adherence to the Heat cured acrylic resin surface. Two-way ANOVA test was carried out to ascertain the level of significance of various observations. After calculating Mean and standard deviation of turbidity degree following results was drawn:

  • Mean of the turbidity degree of contamination after Delmopinol application for Heat-cured acrylic resin specimens was 1440.80 CFU (×10 6 /ml) [Table 2]
  • Mean of the turbidity degree of contamination before Delmopinol application for Heat-cured acrylic resin specimens was 550.85 CFU (×10 6 /ml) [Table 2]
  • The turbidity degree of contamination after Delmopinol application for heat-cured acrylic resin specimens was significantly higher than the mean of the turbidity degree of contamination before Delmopinol application for heat-cured acrylic resin specimens [Table 3].
Table 2: Mean and standard deviation of degree of turbidity


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Table 3: Comparison of groups


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   Discussion Top


Dental plaque as dense, non-calcified bacterial mass so firmly adherent to the tooth surface that they resist wash off by salivary flow and rinsing. A 0.5-1.5 micron thick salivary pellicle is deposited on every surface in the oral cavity, natural or synthetic. [24],[25] The initial stage of plaque formation involves attachments of microorganism to this pellicle surface, [26] followed by adherence of bacteria to each other, mediated by the component of the plaque matrix, causing the microorganism to coalesce and form large bacterial masses.

Candida is most commonly associated with denture plaque. [23],[27],[28],[29],[30] These yeasts are present in the saliva of a majority of denture wearers and display an affinity for adherence to acrylic resin.

Chandra et al. [29] showed that the development of this yeast on acrylic resins happens in 3 distinct stages: Initial stage - up to 11 hours. from the colonization, when some micro-colonies begin to be formed; intermediated stage - from 12 hours to 30 hours after colonization, when extracellular material begin to accumulate over colonies; and maturation stage - from 38 hours to 72 hours after colonization, when Candida colonies become totally involved by the extracellular matrix forming a biofilm. They also concluded that antifungal resistance increases during biofilm development as the extracellular matrix acts as a barrier to action of the antifungal.

Eley [30] in 1999 studied a trial of 0.1% and 0.2% delmopinol hydrochloride mouth rinses as adjuncts to normal oral hygiene has been carried out. The microbiological effects were investigated on plaque collected at 12, 24 and 36 weeks. There were no consistent effects on the microscopical or total counts. However, there was a significant reduction in the proportion of Dextran-producing Streptococci in the active compared with the control group throughout treatment. There was no colonization by Candida or major shift in bacterial composition in the active group nor was there any decrease in susceptibility to Delmopinol. Thus, Delmopinol seems to mediate its plaque inhibitory and anti-inflammatory effects without causing a major shift in bacterial populations apart from the reduction in Dextran-producing Streptococci.

Addy et al. [31] in 2006 with the meta-analyses of studies of 0.2% Delmopinol mouth rinse as an agent to gingival health and plaque control measures and concluded that Delmopinol 0.2% mouth wash is effective as an adjunct measure for reducing plaque burden and indices of gingivitis, whether or not it is used under the supervision.

The Turbidity Degree of contamination after Delmopinol application for Heat-cured acrylic resin specimens was significantly higher than the Mean of the Turbidity Degree of contamination before Delmopinol application for Heat-cured acrylic resin specimens. The results of the present study were consistent with the findings of Addy et al. [31] that 0.2% Delmopinol as a mouth wash reduces plaque on natural teeth surface.

Therefore, it can be concluded that adherence of C. albicans is less with the contamination after Delmopinol application of the acrylic resin specimens as compared with the contamination before Delmopinol application. These findings can be correlated with the finding of Eley [30] that Delmopinol is truly a surfactant agent and has no antibacterial and antifungal effect.

Adherence of microorganisms and debris is favored by rough or otherwise irregular surface topography. [10],[11],[12],[32] Surface irregularities provide an increase in surface area and expansion in the number of niches not readily cleansed by action of tongue or other orofacial musculature. It has been shown that choosing an appropriate type of smooth acrylic resin could lead to reduced biofilm formation. [33]

Though, care has been taken to simulate this in-vitro study to clinical conditions, but variation may be occurring in-vitro. Furthermore, Owing to a small sample size, the result may not truly represent the population and further in-vitro investigation in this field with a larger sample size may be required.


   Conclusion Top


  • Heat-cured acrylic resin shows reduction in adherence of C. albicans by contamination after Delmopinol application
  • Heat-cured acrylic resin shows reduction in adherence of C. albicans by contamination before Delmopinol application
  • Heat-cured acrylic resin shows greater reduction in adherence of C. albicans by contamination after Delmopinol application as compared to contamination before Delmopinol application.


 
   References Top

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Correspondence Address:
Deshraj Jain
Department of Prosthodontics, Government College of Dentistry, Indore, Madhya Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0970-9290.123423

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    Tables

  [Table 1], [Table 2], [Table 3]

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