Effectiveness of chemical disinfection on biofilms of relined dentures: A randomized clinical trial.

PURPOSE: To evaluate the effect of disinfection with sodium perborate or chlorhexidine (when combined with brushing) on the removal of biofilm in relined dentures. METHODS: Swabs were collected 48 hours after the relining procedure and at the follow-up time intervals of 7, 15, 30, 90, and 180 days. The dentures' surface roughness was measured at the same times. 45 subjects were randomly divided into three groups of 15 subjects each. The control group brushed with coconut soap and a soft toothbrush. The sodium perborate group followed the same procedure and also disinfected with sodium perborate solution for 5 minutes per day. The chlorhexidine group followed the control group procedure and disinfected with 2% chlorhexidine digluconate solution for 5 minutes per day. The number of colony forming units and the surface roughness were evaluated statistically by 2-way repeated-measure ANOVA (α = 0.05). RESULTS: The control group dentures exhibited similar levels of microbial cells throughout the experiment. However, after 15 days, no microbial growth was observed on the dentures for which either disinfection agent was used. There were no statistically significant differences in superficial roughness between the groups (P = 0.298). The disinfection agents used, combined with brushing, were able to remove the relined dentures' biofilm after 15 days of disinfection. Roughness was not a predominant factor in CFU reduction.

In Vitro Identification of Histatin 5 Salivary Complexes.

With recent progress in the analysis of the salivary proteome, the number of salivary proteins identified has increased dramatically. However, the physiological functions of many of the newly discovered proteins remain unclear. Closely related to the study of a protein's function is the identification of its interaction partners. Although in saliva some proteins may act primarily as single monomeric units, a significant percentage of all salivary proteins, if not the majority, appear to act in complexes with partners to execute their diverse functions. Coimmunoprecipitation (Co-IP) and pull-down assays were used to identify the heterotypic complexes between histatin 5, a potent natural antifungal protein, and other salivary proteins in saliva. Classical protein-protein interaction methods in combination with high-throughput mass spectrometric techniques were carried out. Co-IP using protein G magnetic Sepharose TM beads suspension was able to capture salivary complexes formed between histatin 5 and its salivary protein partners. Pull-down assay was used to confirm histatin 5 protein partners. A total of 52 different proteins were identified to interact with histatin 5. The present study used proteomic approaches in conjunction with classical biochemical methods to investigate protein-protein interaction in human saliva. Our study demonstrated that when histatin 5 is complexed with salivary amylase, one of the 52 proteins identified as a histatin 5 partner, the antifungal activity of histatin 5 is reduced. We expected that our proteomic approach could serve as a basis for future studies on the mechanism and structural-characterization of those salivary protein interactions to understand their clinical significance.

Histatin 5 inhibits adhesion of C. albicans to Reconstructed Human Oral Epithelium.

Candida albicans is the most pathogenic fungal species, commonly colonizing on human mucosal surfaces. As a polymorphic species, C. albicans is capable of switching between yeast and hyphal forms, causing an array of mucosal and disseminated infections with high mortality. While the yeast form is most commonly associated with systemic disease, the hyphae are more adept at adhering to and penetrating host tissue and are therefore frequently observed in mucosal fungal infections, most commonly oral candidiasis. The formation of a saliva-derived protein pellicle on the mucosa surface can provide protection against C. albicans on oral epithelial cells, and narrow information is available on the mucosal pellicle composition. Histatins are one of the most abundant salivary proteins and presents antifungal and antibacterial activities against many species of the oral microbiota, however, its presence has never been studied in oral mucosa pellicle. The objective of this study was to evaluate the potential of histatin 5 to protect the Human Oral Epithelium against C. albicans adhesion. Human Oral Epithelial Tissues (HOET) were incubated with PBS containing histatin 5 for 2 h, followed by incubation with C. albicans for 1 h at 37°C. The tissues were then washed several times in PBS, transferred to fresh RPMI and incubated for 16 h at 37°C at 5% CO2. HOET were then prepared for histopathological analysis using light microscopy. In addition, the TUNEL assay was employed to evaluate the apoptosis of epithelial cells using fluorescent microscopy. HOET pre-incubated with histatin 5 showed a lower rate of C. albicans growth and cell apoptosis when compared to the control groups (HOET alone and HOET incubated with C. albicans). The data suggest that the coating with histatin 5 is able to reduce C. albicans colonization on epithelial cell surfaces and also protect the basal cell layers from undergoing apoptosis.