Antimicrobial activity of violacein against oral bacteria associated with halitosis: an in vitro study

Introduction: violacein is a natural purple pigment produced by environmental bacteria that presents antimicrobial activity, particularly against Gram-positive bacteria. Intraoral halitosis (IOH) is a condition defined by the unpleasant odor emanating from the mouth, whose main source are volatile sulfur compounds, produced by Gram-negative oral bacteria on the tongue coating. In IOH treatment, antimicrobials have been indicated as chemical adjuncts, including natural products. Objective: thus, this study tested the antimicrobial activity of a violacein extract on key IOH-related bacteria (Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum, Prevotella intermedia, Solobacterium moorei). Materials and Methods: bacteria were cultured in fastidious anaerobe blood agar in anaerobiosis, and 109 cells/ml suspensions were plated. Crude extract of violacein obtained from Chromobacterium violaceum was diluted in a 25% ethanol aqueous solution to 8, 4, 2, 1, 0.5 and 0.25 mg/ml. Using the disk agar diffusion method, 10 µl aliquots of each dilution were deposited on the seeded plates. Chlorohexidine (0.1%) and 25% ethanol solution were used as controls. Plates were incubated in anaerobiosis at 37°C for 72h, and the inhibition halos were recorded. Results: although chlorhexidine showed higher inhibition halos than the violacein extract, most species were inhibited at 4 and 8 mg/ml concentrations (p<0.05). P. gingivalis followed by F. nucleatum were the most affected species in relation to the other bacteria, although statistical significance was only reached for P. gingivalis (p<0.05). Conclusion: crude violacein extract from C. violaceum demonstrated antimicrobial activity against IOH-associated oral bacteria, being a potential antimicrobial to be studied as an adjunct in the control of IOH.

Introdução: a violaceína é um pigmento roxo natural produzido por bactérias ambientais que apresenta ação antimicrobiana, particularmente contra bactérias Gram-positivas. A halitose intraoral (HIO) é uma condição definida pelo odor desagradável que emana da boca, cuja principal fonte são os compostos sulfurados voláteis produzidos por bactérias Gram-negativas da saburra lingual. No tratamento da HIO, antimicrobianos têm sido indicados como adjuvantes, incluindo produtos naturais. Objetivo: assim, este estudo avaliou o potencial antimicrobiano de um extrato de violaceína em patógenos-chave da HIO (Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum, Prevotella intermedia, Solobacterium moorei). Materiais e Métodos: bactérias foram cultivadas em meio ágar sangue para fastidiosos, em anaerobiose, e suspensões de 109 células/ml foram semeadas. O extrato bruto de violaceína obtido de Chromobacterium violaceum foi diluído em solução aquosa com 25% de etanol nas concentrações de 8, 4, 2, 1, 0,5 e 0,25 mg/ml. Através do método de disco difusão, 10 µl de cada diluição foram depositados nas placas semeadas. A clorexidina (0,1%) e a solução etanólica a 25% foram usadas como controles. As placas foram incubadas em anaerobiose a 37°C por 72h, e os halos de inibição foram registrados. Resultados: embora a clorexidina tenha apresentado os maiores halos de inibição do do que o extrato, a maioria das espécies foi inibida nas concentrações de 4 e 8 mg/ml (p<0,05). P. gingivalis e F. nucleatum foram as espécies mais afetadas em relação às outras bactérias, porém só foi observada significância estatística para P. gingivalis (p<0,05). Conclusão: o extrato bruto de violaceína de C. violaceum demonstrou atividade antimicrobiana contra bactérias orais associadas a HIO, sendo um potencial antimicrobiano a ser estudado como adjuvante no controle da HIO.

Phototoxic assessment of a sunscreen formulation and its excipients: An in vivo and in vitro study.

BACKGROUND: Cosmetic preservatives are used to protect cosmetic formulations and improve its shelf-life. However, these substances may exert phototoxic effects when used under sunlight. OBJECTIVE: To assess safety, efficacy and putative phototoxic effects of a sunscreen formulation SPF 30 and its excipients. MATERIALS/METHODS: Irradiation was performed with solar simulated light (SSL) and the sunscreen from the School of Pharmacy/UFRJ/Brazil. We used albino hairless mice in different groups (control (G1), only irradiated (G2), sunscreen plus irradiation (G3) and vehicle plus irradiation (G4) for morphological assessment and immunefluorescence detection to OKL38. In vitro analyses were with a Saccharomyces cerevisiae (SC) strain plus SSL in the presence of methylparaben, propylparaben, imidazolidinyl urea, aminomethyl propanol and their association. RESULTS: G3 and G4 displayed photosensitization leading to thickening of the epidermis and increased dermal cellularity. G4 displayed strong OKL38 labeling when compared with other groups. Aminomethyl propanol, methylparaben and propylparaben are endowed with phototoxic activity against SC. Propylparaben displayed the highest phototoxic effect, followed by excipients association. CONCLUSIONS: The sunscreen's vehicle is endowed with phototoxic activity. Propylparaben was the most phototoxic agent, increasing the overall phototoxicity of excipient association, pointing to a critical concern regarding vehicle associations intended to cosmetic purposes.