Dual nanocarrier of chlorhexidine and fluconazole: Physicochemical characterization and effects on microcosm biofilms and oral keratinocytes.

OBJECTIVES: This study assembled and characterized a dual nanocarrier of chlorhexidine (CHX) and fluconazole (FLZ), and evaluated its antibiofilm and cytotoxic effects. METHODS: CHX and FLZ were added to iron oxide nanoparticles (IONPs) previously coated by chitosan (CS) and characterized by physical-chemical analyses. Biofilms from human saliva supplemented with Candida species were grown (72 h) on glass discs and treated (24 h) with IONPs-CS carrying CHX (at 39, 78, or 156 µg/mL) and FLZ (at 156, 312, or 624 µg/mL) in three growing associations. IONPs and CS alone, and 156 µg/mL CHX + 624 µg/mL FLZ (CHX156-FLZ624) were tested as controls. Next, microbiological analyses were performed. The viability of human oral keratinocytes (NOKsi lineage) was also determined (MTT reduction assay). Data were submitted to ANOVA or Kruskal-Wallis, followed by Fisher's LSD or Tukey's tests (α=0.05). RESULTS: Nanocarriers with spherical-like shape and diameter around 6 nm were assembled, without compromising the crystalline property and stability of IONPs. Nanocarrier at the highest concentrations was the most effective in reducing colony-forming units of Streptococcus mutans, Lactobacillus spp., Candida albicans, and Candida glabrata. The other carriers and CHX156-FLZ624 showed similar antibiofilm effects, and significantly reduced lactic acid production (p<0.001). Also, a dose-dependent cytotoxic effect against oral keratinocytes was observed for the dual nanocarrier. IONPs-CS-CHX-FLZ and CHX-FLZ significantly reduced keratinocyte viability at CHX and FLZ concentrations ≥7.8 and 31.25 µg/mL, respectively (p<0.05). CONCLUSION: The nanotherapy developed outperformed the effect of the combination CHX-FLZ on microcosm biofilms, without increasing the cytotoxic effect of the antimicrobials administered. CLINICAL SIGNIFICANCE: The dual nanocarrier is a promising topically-applied therapy for the management of oral candidiasis considering that its higher antibiofilm effects allow the use of lower concentrations of antimicrobials than those found in commercial products.

Nanocarriers of Miconazole or Fluconazole: Effects on Three-Species Candida Biofilms and Cytotoxic Effects In Vitro.

The contribution of different Candida species in oral fungal infections has stimulated the search for more effective therapies. This study assessed the antibiofilm effects of nanocarriers of miconazole (MCZ) or fluconazole (FLZ) on Candida biofilms, and their cytotoxic effects on murine fibroblasts. Three-species biofilms (Candida albicans/Candida glabrata/Candida tropicalis) were formed on 96-well plates, and they were treated with nanocarriers (iron oxide nanoparticles coated with chitosan-"IONPs-CS") of MCZ or FLZ at 39/78/156 µg/mL; antifungals alone at 156 µg/mL and artificial saliva were tested as positive and negative controls, respectively. Biofilms were analyzed by colony forming units (CFU), biomass, metabolic activity, and structure/viability. The cytotoxicity (L929 cells) of all treatments was determined via 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) reduction assay. Data were submitted to one- or two-way ANOVA, followed by Tukey's or Fisher LSD's tests (p < 0.05). IONPs-CS-MCZ at 78 µg/mL promoted similar antibiofilm and cytotoxic effects compared with MCZ at 156 µg/mL. In turn, IONPs-CS-FLZ at 156 µg/mL was overall the most effective FLZ antibiofilm treatment, surpassing the effects of FLZ alone; this nanocarrier was also less cytotoxic compared with FLZ alone. It can be concluded that both nanocarriers are more effective alternatives to fight Candida biofilms compared with their respective positive controls in vitro, being a promising alternative for the treatment of oral fungal infections.

Effects of Antifungal Carriers Based on Chitosan-Coated Iron Oxide Nanoparticles on Microcosm Biofilms.

Resistance of Candida species to conventional therapies has motivated the development of antifungal nanocarriers based on iron oxide nanoparticles (IONPs) coated with chitosan (CS). This study evaluates the effects of IONPs-CS as carriers of miconazole (MCZ) or fluconazole (FLZ) on microcosm biofilms. Pooled saliva from two healthy volunteers supplemented with C. albicans and C. glabrata was the inoculum for biofilm formation. Biofilms were formed for 96 h on coverslips using the Amsterdam Active Attachment model, followed by 24 h treatment with nanocarriers containing different concentrations of each antifungal (78 and 156 µg/mL). MCZ or FLZ (156 µg/mL), and untreated biofilms were considered as controls. Anti-biofilm effects were evaluated by enumeration of colony-forming units (CFUs), composition of the extracellular matrix, lactic acid production, and structure and live/dead biofilm cells (confocal laser scanning microscopy-CLSM). Data were analyzed by one-way ANOVA and Fisher LSD's test (α = 0.05). IONPs-CS carrying MCZ or FLZ were the most effective treatments in reducing CFUs compared to either an antifungal agent alone for C. albicans and MCZ for C. glabrata. Significant reductions in mutans streptococci and Lactobacillus spp. were shown, though mainly for the MCZ nanocarrier. Antifungals and their nanocarriers also showed significantly higher proportions of dead cells compared to untreated biofilm by CLSM (p < 0.001), and promoted significant reductions in lactic acid, while simultaneously showing increases in some components of the extracellular matrix. These findings reinforce the use of nanocarriers as effective alternatives to fight oral fungal infections.

Oral prosthetic microbiology: aspects related to the oral microbiome, surface properties, and strategies for controlling biofilms.

The oral cavity is an environment that allows for the development of complex ecosystems; the placement of prosthetic devices as a consequence of partial or total tooth loss may alter the diversity of microbial communities. Biofilms on the surface of materials used in dental prostheses can promote important changes in the mechanic and aesthetic properties of the material itself and may cause local and systemic diseases for the prosthetic wearer. This review presents the main features of the oral microbiome associated with complete or partial dentures and dental implants. The main diseases associated with microbial colonization of prosthetic surfaces, factors that may affect biofilm formation on prosthetic materials, as well as novel alternative therapies aiming to reduce biofilm formation and/or to eradicate biofilms formed on these materials are also explored.

Novel Colloidal Nanocarrier of Cetylpyridinium Chloride: Antifungal Activities on Candida Species and Cytotoxic Potential on Murine Fibroblasts.

Nanocarriers have been used as alternative tools to overcome the resistance of Candida species to conventional treatments. This study prepared a nanocarrier of cetylpyridinium chloride (CPC) using iron oxide nanoparticles (IONPs) conjugated with chitosan (CS), and assessed its antifungal and cytotoxic effects. CPC was immobilized on CS-coated IONPs, and the nanocarrier was physico-chemically characterized. Antifungal effects were determined on planktonic cells of Candida albicans and Candida glabrata (by minimum inhibitory concentration (MIC) assays) and on single- and dual-species biofilms of these strains (by quantification of cultivable cells, total biomass and metabolic activity). Murine fibroblasts were exposed to different concentrations of the nanocarrier, and the cytotoxic effect was evaluated by MTT reduction assay. Characterization methods confirmed the presence of a nanocarrier smaller than 313 nm. IONPs-CS-CPC and free CPC showed the same MIC values (0.78 µg mL-1). CPC-containing nanocarrier at 78 µg mL-1 significantly reduced the number of cultivable cells for all biofilms, surpassing the effect promoted by free CPC. For total biomass, metabolic activity, and cytotoxic effects, the nanocarrier and free CPC produced statistically similar outcomes. In conclusion, the IONPs-CS-CPC nanocarrier was more effective than CPC in reducing the cultivable cells of Candida biofilms without increasing the cytotoxic effects of CPC, and may be a useful tool for the treatment of oral fungal infections.

Profundidade do sulco gengival em pacientes de faixa etária de 18-25 anos / Depth of gingival sulcus in patients aged between 18 to 25 years

Gingival probing performed in periodontal normal condition brings important results for the definition of health and diseases of the periodontal tissues. The aim was to evaluate the depth probing of the gingival sulcus in permanent dentition of young adults. It was carried out a transversal study with 120 volunteers aged 18 to 25 years and with healthy gum. It was used a manual periodontal probe Williams model. The evaluation was performed on all four sides, with a total of four measurements per tooth. The collected data were first tabulated and submitted to Kruskal-Wallis test with Dunn's post test.The mean and standard deviation of the upper teeth were: incisor (1.1343 ± 0.33665); canine (1.1819 ± 0.369); premolar ratio (1.3677 ± 0.3752); molar(1.8030 ± 0.4116). The mean of the lower teeth was: incisor (1.1260 ± 0.3272); canine (1.2106 ± 0.3390); premolar ratio (1.4580 ± 0.3778); molar (1.9068 ± 0.4497).It concludes that the average depth of the gingival sulcus, between the group of teeth was higher in the lower arch relative to the upper; except for faces: incisive distal, buccal molar and the lingual incisor and canine; in which the result related to the average depth is less than the upper arch. (AU)

A sondagem gengival realizada em condições periodontais normais traz importantes resultados para a definição de saúde e doenças dos tecidos periodontais. O objetivo deste estudo foi avaliar a profundidade clínica do sulco gengival em uma dentição jovem. Um estudo transversal foi realizado com 120 voluntários entre 18 e 25 anos com gengiva saudável. Um modelo de Williams de sonda periodontal manual foi utilizado. A avaliação foi realizada nos quatro lados, com um total de quatro medidas por dente. Os dados coletados foram tabulados e submetidos ao teste de KruskalWallis com pós-teste de Dunn. A média e o desvio padrão dos dentes superiores foram: incisivo (1,1343 ± 0,33665); canino (1,1819 ± 0,369); pré molar (1,3677 ± 0,3752); molar (1,8030 ± 0,4116). A média dos dentes inferiores foi: incisivo (1,1260 ± 0,3272); canino (1,2106 ± 0,3390); pré-molar (1,4580 ± 0,3778); molar (1,9068 ± 0,4497). Concluise que a profundidade média do sulco gengival, entre o grupo de dentes, foi maior no arco inferior em relação ao superior; exceto por faces: incisiva distal, molar vestibular e incisivo lingual e canino; em que o resultado relacionado à profundidade média é menor que o arco superior (AU)