Improving Drug Delivery on Candida Albicans Using Geraniol Nanoemulsion.

Geraniol (GE) is a monoterpene alcohol with excellent antifungal activity. However, its low solubility and high volatility impair its use. Nanoemulsions (NE) are excellent delivery systems for poorly soluble and volatile drugs, achieving controlled release of the active ingredient. The aim of this study was to improve the delivery of geraniol (GE) incorporated in NE against Candida albicans in order to evaluate the antibiofilm effect and cytotoxicity. Nanoemulsion containing 10% oil phase (cholesterol) (w/w), 10% surfactant (mixture of soy phosphatidylcholine and Brij 58; 1:2) (w/w), and 80% aqueous phase (phosphate buffer) (w/w) was synthesized. Incorporation of GE was carried out by sonication and the final compounds were characterized by hydrodynamic diameter, polydispersity index (PDI), and zeta potential (ZP), in addition to evaluation of physicochemical stability after 6 months and 1 year. The GE-NE effect was evaluated on Candida albicans biofilms and cytotoxic effect was evaluated on immortalized normal oral cell line NOK-Si. The diameter of GE-NE was 232.3 ± 2.7 nm and PDI 0.155 with exhibited homogeneity and stability in solution. GE-NE showed antibiofilm activity at a concentration of 75 µg/mL with reduction of >6.0 log10, and no cytotoxicity against NOK-Si cells at concentrations below 150 µg/mL was observed. GE-NE proved to be a promising candidate for prevention and treatment of fungal diseases.

Chlorin-e6 conjugated to the antimicrobial peptide LL-37 loaded nanoemulsion enhances photodynamic therapy against multi-species biofilms related to periodontitis.

In our previous studies, Chlorin-e6 (Ce6) demonstrated a significant reduction of microorganisms' viability against multi-species biofilm related to periodontitis while irradiated with blue light. However, the conjugation of Ce6 and antimicrobial peptides, and the incorporation of this photosensitizer in a nanocarrier, is still poorly explored. We hypothesized that chlorin-e6 conjugated to the antimicrobial peptide LL-37 loaded nanoemulsion could inhibit a multi-species biofilm related to periodontitis during photodynamic therapy (PDT), the pre-treatment with hydrogen peroxide was also tested. The nanoemulsion (NE) incorporated with Ce6 was characterized regarding the physiochemical parameters. Images were obtained by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Later, the Ce6 and LL-37 incorporated in NE was submitted to UV-Vis analysis and Reactive Oxygen Species (ROS) assay. Finally, the combined formulation (Ce6+LL-37 in nanoemulsion) was tested against multi-species biofilm related to periodontitis. The formed nanoformulation was kinetically stable, optically transparent with a relatively small droplet diameter (134.2 unloaded and 146.9 loaded), and weak light scattering. The NE system did not impact the standard UV-VIS spectra of Ce6, and the ROS production was improved while Ce6 was incorporated in the NE. The combination of Ce6 and LL-37 in NE was effective to reduce the viability of all bacteria tested. The treatment with hydrogen peroxide previous to PDT significantly impacted bacterial viability. The current aPDT regimen was the best already tested against periodontal biofilm by our research team. Our results suggest that this combined protocol must be exploited for clinical applications in localized infections such as periodontal disease. - Nanoemulsion demonstrated to be an excellent nanocarrier for photodynamic application. - Chlorin-e6 incorporated in nanoemulsion showed great physicochemical and biophotonic parameters. - The combination of chlorin-e6 and LL-37 peptide in nanoemulsion is effective to eliminate periodontal pathogenic bacteria. - The treatment with hydrogen peroxide previous to PDT significantly impacted bacterial viability.

Chlorin-based photosensitizer under blue or red-light irradiation against multi-species biofilms related to periodontitis.

In our previous study, Chlorin-e6 (Ce6) demonstrated a significant reduction of microorganisms' viability against single-species biofilm related to periodontitis once irradiated by red light (660 nm). Also, higher bacteria elimination was observed under blue light (450 nm) irradiation. However, the use of blue light irradiation of Ce6 for antimicrobial administration is poorly explored. This study evaluated the effect of chlorin-e6-mediated antimicrobial photodynamic therapy (aPDT) using different wavelengths (450 or 660 nm) against multi-species biofilms related to periodontitis. Streptococcus oralis, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans composed the mature biofilm developed under proper conditions for five days. aPDT was performed using different concentrations of Ce6 (100 and 200 µM), wavelengths (450 or 660 nm), and comparisons were made after qPCR assay and confocal laser scanning microscopy (CLSM) analysis. The greatest bacterial elimination was observed in the groups where Ce6 was used with blue light, for S. orallis (2.05 Log10 GeQ mL-1, p < 0.0001) and P. gingivalis (1.4 Log10 GeQ mL-1, p < 0.0001), aPDT with red light showed significant bacteria reduction only for S. orallis. aPDT with blue light demonstrated statistically higher elimination in comparison with aPDT with red light. The aPDT did not show a statistically significant effect when tested against A. actinomycetemcomitans and F. nucleatum (p=0.776 and 0.988, respectively). The aPDT using blue light showed a promising higher photobiological effect, encouraging researchers to consider it in the irradiation of Ce6 for further investigations.

Photodynamic inactivation using a chlorin-based photosensitizer with blue or red-light irradiation against single-species biofilms related to periodontitis.

Chlorin-e6 (Ce6), as a photosensitizer (PS), has demonstrated significant reduction of microorganisms' viability when irradiated by red light. However, the main absorption peak of this PS is located at blue light spectrum, which is less investigated. This study aimed to evaluate the effect of pure-chlorin-e6-mediated photodynamic inactivation (PDI) using different light sources (450 or 660 nm) against biofilms related to periodontitis. Streptococcus oralis, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans single-species biofilms were developed under proper conditions for five days. PDI was performed using different concentrations of Ce6 (100 and 200 mM), wavelengths (450 and 660 nm) and comparisons were made after colony forming unit and confocal laser scanning microscopy (CLSM) analysis. The use of light and PS were also individually tested. The greatest bacterial elimination was observed in the group where PDI was employed with blue light and concentration of 200 mM for all bacterial strains tested (4.01 log10 for A. actinomycetemcomitans, and total elimination for P. gingivalis and S. oralis), except for F. nucleatum, where 3.46 log10 reduction was observed when red light and 200 mM Ce6 were applied (p < 0.05). The antimicrobial effects of PDI mediated by Ce6 for all single pathogenic biofilms were confirmed by live/dead staining under CLSM analysis. For all single-species biofilms, the use of PDI mediated by chlorin-e6 photosensitizer under blue or red-light irradiation (450 and 660 nm) demonstrated a significant reduction in bacterial viability, but blue light showed a promising higher photobiological effect, encouraging its adjuvant use to basic periodontitis treatment.

Terpinen-4-ol and carvacrol affect multi-species biofilm composition.

The aim of this study was to investigate the cytotoxic activity and inhibitory effect of terpinen-4-ol (T4ol) and carvacrol against single- and multi-species biofilms. The toxicity of each compound was tested on oral keratinocytes and evaluated by XTT assay. Inhibition and eradication of single-species biofilms were analyzed by crystal violet assay and the effect on multi-species biofilm composition was evaluated by qPCR. T4ol and carvacrol did not affect the epithelial cell viability, in contrast to chlorhexidine, which showed a high cytotoxic effect. Inhibition and eradication of single-species biofilms treated with T4ol and carvacrol were observed. The same inhibitory effect was observed for multi-species biofilms, especially on periodontal pathogens. In conclusion, specific concentrations of T4ol and carvacrol without toxicity towards the epithelial cells reduced the numbers of periodontal pathogens in single- and multi-species biofilms.

Analysis of the biofilm formed on the surface of a glass-ionomer cement associated with different concentrations of chlorhexidine diacetate.

PURPOSE: This cross-over in situ experiment evaluated the microbial composition and quantified the total polysaccharides in the biofilm formed on the surface of a high viscosity glass-ionomer cement (GIC) mixed with chlorhexidine diacetate (CHX) in concentrations of 0.5%, 1% and 2%. GIC without CHX was used as control. METHODS: The volunteers (n= 8) tested each material for a 7-day period, by using a palatal acrylic device containing four round specimens, all fabricated with the same material. An increasing order of CHX concentration and a 15-day washout period between each concentration was adopted. The biofilm formed was collected and inoculated in specific culture media and thereafter, the total microorganisms, total streptococci, Streptococcus mutans and Lactobacillus spp were counted. Total polysaccharides were quantified by using a phenol-sulfuric method. Microbiologic data were analyzed by ANOVA for repeated measures and Tukey tests, and the total polysaccharide content by Friedman and Dunn's tests. Significance level was set at 5%. RESULTS: Microorganism counts showed statistically significant differences among groups only for Lactobacillus (P< 0.05), that were significantly higher for group GIC+CHX 2% compared with GIC and GIC+CHX 0.5%. However, the concentration of total polysaccharides in the biofilm was lower for groups containing 1% and 2% of CHX. CLINICAL SIGNIFICANCE: The addition of chlorhexidine in the studied concentrations did not reduce the microorganism counts, but caused reduction of biofilm density.

Aesthetic rehabilitation of oligodontia in primary dentition with adhesive partial denture.

The primary teeth are essential for bone development and establishment of the arches on occlusion. Thus, the congenitally absence of teeth may trigger a shift in the balance of the occlusion, promoting disharmony in the structures of the maxilla-mandibular system. However, some interventions are possible to be performed in these cases even in pediatric patients, to redirect growth, preventing growth deviations and reestablishing the aesthetic. The aim of this paper is to report the treatment of a 4-year-old child presenting congenitally absence of mandibular central and lateral incisors and maxilla lateral incisors, which consequently compromises aesthetics, occlusal function, and the development and the functional growth of the bones. The oral rehabilitation was performed with an adhesive partial denture, which was able to restore the aesthetic and the occlusal function, therefore being a viable alternative in the treatment of this patient of little age.

Implications of Surface and Bulk Properties of Abutment Implants and Their Degradation in the Health of Periodontal Tissue.

The aim of the current review was to investigate the implications of the surface and bulk properties of abutment implants and their degradation in relation to periodontal health. The success of dental implants is no longer a challenge for dentistry. The scientific literature presents several types of implants that are specific for each case. However, in cases of prosthetics components, such as abutments, further research is needed to improve the materials used to avoid bacterial adhesion and enhance contact with epithelial cells. The implanted surfaces of the abutments are composed of chemical elements that may degrade under different temperatures or be damaged by the forces applied onto them. This study showed that the resulting release of such chemical elements could cause inflammation in the periodontal tissue. At the same time, the surface characteristics can be altered, thus favoring biofilm development and further increasing the inflammation. Finally, if not treated, this inflammation can cause the loss of the implant.

Porphyromonas gingivalis-mediated shedding of extracellular matrix metalloproteinase inducer (EMMPRIN) by oral epithelial cells: a potential role in inflammatory periodontal disease.

Extracellular matrix metalloproteinase inducer (EMMPRIN) or CD147 is a transmembrane glycoprotein expressed by various cell types, including oral epithelial cells. Recent studies have brought evidence that EMMPRIN plays a role in periodontitis. In the present study, we investigated the effect of Porphyromonas gingivalis, a major pathogen in chronic periodontitis, on the shedding of membrane-anchored EMMPRIN and on the expression of the EMMPRIN gene by oral epithelial cells. A potential contribution of shed EMMPRIN to the inflammatory process of periodontitis was analyzed by evaluating the effect of recombinant EMMPRIN on cytokine and matrix metalloproteinase (MMP) secretion by human gingival fibroblasts. ELISA and immunofluorescence analyses revealed that P. gingivalis mediated the shedding of epithelial cell-surface EMMPRIN in a dose- and time-dependent manner. Cysteine proteinase (gingipain)-deficient P. gingivalis mutants were used to demonstrate that both Arg- and Lys-gingipain activities are involved in EMMPRIN shedding. Real-time PCR showed that P. gingivalis had no significant effect on the expression of the EMMPRIN gene in epithelial cells. Recombinant EMMPRIN induced the secretion of IL-6 and MMP-3 by gingival fibroblasts, a phenomenon that appears to involve mitogen activated protein kinases. The present study brought to light a new mechanism by which P. gingivalis can promote the inflammatory response during periodontitis.

El papel del óxido nítrico en la modulación del proceso inflamatorio de la enfermedad periodontal / The role of nitric oxide in the modulation of inflammatory periodontal disease

El óxido nítrico (NO) es un radical libre que participa de prácticamente todos los procesos fisiológicos del organismo; sin embargo, en grandes concentraciones puede actuar de forma perjudicial en los tejidos. Su acción inmuno-regulatoria está presente en la inflamación y en los mecanismos de auto inmunidad, siendo intensamente estudiada en el área médica. Recientemente, algunos estudios también han citado al NO como factor etiopatogénico de la enfermedad periodontal, el cual posee naturaleza inflamatoria y multifactorial. La acción del NO depende de su concentración en el organismo, pudiendo ser benéfica o perjudicial y, basada en eso, investigaciones han enfocado el efecto de algunos inhibidores de NO como supuestos agentes terapéuticos en los procesos inflamatorios. A partir de ese contexto, este trabajo tuvo como objetivo relatar el papel del NO y de la inhibición de la óxido nítrico sintasa (NOS) en el proceso de la modulación de la enfermedad periodontal. En conclusión, se puede sugerir que el NO parece ejercer un papel fundamental en la evolución de la enfermedad periodontal inflamatoria y, que la inhibición de la NOS puede ser considerada una terapéutica promisora en la modulación del proceso

Nitric oxide (NO) is a free radical with participation in almost all physiologic host processes; however, in high concentrations it may damage the tissues. Its immunoregulatory action is present in the inflammation and in auto-immune mechanisms, being intensively studied in the medical area. Recently, some studies have also reported that NO could play a role as etiopathogenic factor of the periodontal disease, which shows inflammatory and multifactorial course. Due to beneficial or damage effects of NO, according to its concentrations, some studies have been focused in the evaluation of inhibitor of NO-Synthase (NOS) as therapeutic agents in inflammatory processes. In this context, the aim of this study was to report the role of NO and NOS inhibition in the periodontal disease modulation process. In conclusion, it could be suggested that NO seems to play an essential role in evolution of inflammatory periodontal disease, and that the NOS inhibition may be considered as a promising therapeutic in modulation of inflammatory process

Denture disinfection by microwave irradiation: a randomized clinical study.

OBJECTIVE: This study evaluated the clinical effectiveness of two exposure times of microwave irradiation on the disinfection of complete dentures. METHODS: Biofilm samples were collected from dentures of 30 patients, who were randomly divided into two experimental groups of 15 subjects each: Group 1-patients had their maxillary denture microwaved for 3 min (650W); Group 2-patients had their maxillary denture microwaved for 2 min (650W). Denture biofilm samples were taken with swabs, before (left side surfaces) and after (right side surfaces) microwave irradiation. All microbial material was plated on selective media for Candida spp., Staphylococcus spp., mutans streptococci and a non-selective media. After incubation (48 h/37 degrees C), the number of colony-forming units (cfu/mL) was counted. Microorganisms which grew on selective media were identified using biochemical methods. The data were statistically analyzed by Kruskal-Wallis test, followed by Dunn's post-test (alpha=0.05). RESULTS: Microwave irradiation for 3 min (Group 1) resulted in sterilization of all dentures evaluated. After microwave irradiation for 2 min (Group 2), a significant decrease in Candida spp. (P=0.0062), Staphylococcus spp. (P=0.0178), mutans streptococci (P=0.0047) and non-identified species (P<0.0001) was achieved in comparison with the cfu/mL obtained before irradiation. The colonies grown after 2 min of microwave irradiation were identified as Candida albicans, non-aureus Staphylococci and Streptococcus mutans. CONCLUSION: Microwave irradiation for 3 min may be a potential treatment to prevent cross-contamination.