Effectiveness of a tissue conditioner containing antifungals in a rat model of denture stomatitis.

AIM: This study investigated the effectiveness of a drug-modified tissue conditioner in an animal model of denture stomatitis. METHODS AND RESULTS: Wistar rats wore a Candida albicans-contaminated palatal device for 4 days. Next, nystatin (Nys) or chlorhexidine (Chx) were added to a tissue conditioner in their raw or ß-cyclodextrin-complexed (ßCD) forms at their minimum inhibitory concentrations. As controls, one group was not subjected to any procedure (NC), one group used sterile devices, one group had denture stomatitis but was not treated (DS), and another had the devices relined with the tissue conditioner without the addition of any drug (Soft). After 4 days of treatment, treatment effectiveness was assessed visually, histologically, and through CFU count, and myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) assays. Rats from the Soft, Nys, Nys:ßCD, and Chx groups presented a significant decrease in the microbial load compared with the untreated group. Treatment groups showed lower MPO and NAG activity compared to the non-treated group. CONCLUSIONS: The addition of antifungals to a soft tissue conditioner can be a promising approach for denture stomatitis treatment.

A novel rat model of denture stomatitis and the role of antibiotics in the development of the disease.

This study compared different conditions to establish a rat model of denture stomatitis. Immunocompetent Wistar rats were divided into two groups (n = 35): Tetracycline = administration of 0.83 mg/ml of tetracycline hydrochloride 7 days before induction of denture stomatitis and amoxicillin = administration of 0.156 mg/ml of amoxicillin with clavulanic acid 4 days before induction of denture stomatitis. A suspension of Candida albicans was inoculated on the palate followed by the use of a palatal device contaminated with C. albicans inoculum for 4 days to induce denture stomatitis. As controls, some rats were not submitted to any procedure or used a sterile palatal device for 4 days. The development of denture stomatitis was confirmed by visual analysis, colony-forming units per milliliter (CFU/ml) count, histopathological and immunohistochemical analyses, and through myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) assays. Rats were euthanized right after device removal (T0), 4 (T4), or 6 (T6) days after device removal. Tetracycline improved the development of the disease, with more severe clinical signs at T0. Similar results were observed in the CFU/ml count and in the histometric and immunohistochemical analyses. Higher MPO expression was detected in the palates of the tetracycline group (P = .006). Despite the subtle differences between antibiotics, tetracycline showed better results in inducing and maintaining denture stomatitis for at least 4 days after device removal.

Denture stomatitis is an oral inflammatory disease with high recurrence rates. Different animal models have been reported in the literature, but some gaps still need to be addressed. A reproducible in vivo model should be established to test new treatment approaches.

Experimental animal models for denture stomatitis: A methodological review.

Denture stomatitis is the most prevalent form of oral candidiasis and the most frequent oral lesion in removable prosthesis wearers. It is characterized by an inflammatory response of the denture-bearing mucosa, especially the palatal mucosa, and its clinical signs include chronic edema and erythema, and papillary hyperplasia. Despite having a multifactorial etiology, its main etiological agent is the infection by Candida albicans. Given its high treatment failure rates, an in vivo model of denture stomatitis should be established to test alternative treatments. The aim of this study is to review the existing denture stomatitis models and to provide an overview of the main methodological differences between them. Over the last 40 years, different animal models were developed in order to study denture stomatitis etiopathogenesis and to assess novel therapies. Many approaches, including the use of antibiotics and immunosuppressors, have to be further investigated in order to establish which protocol is more appropriate and effective for the development of an animal model of denture stomatitis.

Is there an optimal method to detach Candida albicans biofilm from dental materials?

Introduction. Candida albicans can produce a complex, dynamic and resistant biofilm on the surface of dental materials, especially denture base acrylic resins and temporary soft liners. This biofilm is the main aetiological factor for denture stomatitis, an oral inflammatory condition characterized by chronic and diffuse erythema and oedema of the denture bearing mucosa.Gap Statement. There is no consensus in the literature regarding the best method to detach biofilms from dental materials. In order to assess the antifungal efficacy of new materials and treatments, the biofilm needs to be properly detached and quantified.Aim. This study compared different methods of detaching C. albicans biofilm from denture base acrylic resin (Vipi Cril) and temporary soft liner (Softone) specimens.Methodology. Specimens of each material were immersed in an inoculum of C. albicans SC5314 and remained for 90 min in orbital agitation at 75 r.p.m. and 37 °C. After the removal of non-adherent cells, the specimens were immersed in RPMI-1640 medium for 48 h. Biofilm formation was evaluated with confocal laser scanning microscopy (n=5). Then, other specimens (n=7) were fabricated, contaminated and immersed in 3 ml of sterile phosphate-buffered saline (PBS) and vortexed or sonicated for 1, 2, 5, or 10 min to detach the biofilm. The quantification of detached biofilm was performed by colony-forming unit (c.f.u.) ml-1 count. Results were submitted to one-way analysis of variance (ANOVA)/Tukey HSD test (α=0.05).Results. A mature and viable biofilm was observed on the surfaces of both materials. For both materials, there was no significant difference (P>0.05) among detachment methods.Conclusion. Any of the tested methods could be used to detach C. albicans biofilm from hard and soft acrylic materials.

A novel acrylic resin palatal device contaminated with Candida albicans biofilm for denture stomatitis induction in Wistar rats.

Denture stomatitis is the most frequent oral lesion in removable prosthesis wearers, with high recurrence rates and a complex treatment. OBJECTIVE: This study describes a protocol to obtain and to contaminate a palatal device with Candida albicans biofilm that could be used for an animal model of denture stomatitis. METHODOLOGY: Acrylic resin devices (N=41) were obtained from impressions of the palates of Wistar rats with individual trays and polyether. The efficacy of microwave irradiation (MW), ultraviolet light (UV), or ultrasonic bath (US) was assessed by colony viability and spectrophotometric analyses (n=5) in order to select the most appropriate method for sterilizing the devices. Then, different devices (n=5) were contaminated with C. albicans and evaluated by CFU/mL determination, scanning electron microscopy, and laser confocal microscopy. Device stabilization was assessed with either autopolymerizing acrylic resins or a self-adhesive resin cement (n=2). The spectrophotometric data were analyzed by one-way ANOVA followed by the Tukey's HSD post-hoc test (α=0.05). RESULTS: MW was the only method capable of sterilizing the devices, and the contamination protocol developed a mature and viable C. albicans biofilm (~1.2 x 106 CFU/mL). The self-adhesive resin cement was the best stabilization material. CONCLUSIONS: This acrylic resin palatal device was designed to be similar to the clinical situation of contaminated prostheses, with easy manufacturing and handling, effective stabilization, and satisfactory contamination. Thus, the acrylic device can be a valuable tool in the development of denture stomatitis in rats.

A novel acrylic resin palatal device contaminated with Candida albicans biofilm for denture stomatitis induction in Wistar rats

Abstract Denture stomatitis is the most frequent oral lesion in removable prosthesis wearers, with high recurrence rates and a complex treatment. Objective This study describes a protocol to obtain and to contaminate a palatal device with Candida albicans biofilm that could be used for an animal model of denture stomatitis. Methodology Acrylic resin devices (N=41) were obtained from impressions of the palates of Wistar rats with individual trays and polyether. The efficacy of microwave irradiation (MW), ultraviolet light (UV), or ultrasonic bath (US) was assessed by colony viability and spectrophotometric analyses (n=5) in order to select the most appropriate method for sterilizing the devices. Then, different devices (n=5) were contaminated with C. albicans and evaluated by CFU/mL determination, scanning electron microscopy, and laser confocal microscopy. Device stabilization was assessed with either autopolymerizing acrylic resins or a self-adhesive resin cement (n=2). The spectrophotometric data were analyzed by one-way ANOVA followed by the Tukey's HSD post-hoc test (α=0.05). Results MW was the only method capable of sterilizing the devices, and the contamination protocol developed a mature and viable C. albicans biofilm (~1.2 x 106 CFU/mL). The self-adhesive resin cement was the best stabilization material. Conclusions This acrylic resin palatal device was designed to be similar to the clinical situation of contaminated prostheses, with easy manufacturing and handling, effective stabilization, and satisfactory contamination. Thus, the acrylic device can be a valuable tool in the development of denture stomatitis in rats.

Candida-Associated Denture Stomatitis and Murine Models: What Is the Importance and Scientific Evidence?

Considering the high prevalence and recurrence of Candida-associated denture stomatitis (CADS), in vivo studies in animal models are necessary before those in humans to evaluate new therapeutic strategies. This study aimed to review the literature on murine models of CADS induction using acrylic intraoral devices simulating dentures. Rats are recommended as experimental animals in these models as well as the adoption of a pasty diet. For maintenance in the proper position during the experiments, intraoral appliances must be obtained by individual impressions, using and retained exclusively by cementation on the molars. The region of interest for histopathological analysis was standardized as that corresponding to the area between the first molars. However, there is no consensus among the studies on the CADS induction rat models in relation to the Candida albicans inoculation and need for immunosuppression and/or administration of antibacterial drugs of animals. The greatest difficulty of the available models refers to maintaining the course of the lesion for a sufficient period to evaluate the effectiveness of the proposed treatment, considering the rapid and efficient murine immune response to candidal colonization. Therefore, future studies are necessary for the development of a robust and reproducible CADS model.