phytochlorin and Periodontal-Diseases

The antimicrobial photodynamic therapy (aPDT) has stood out as an alternative and promising method of disinfection and has been exploited for the treatment of oral bacteria. In this study, we evaluate in vitro the action of aPDT, mediated by methylene blue, chlorin-e6, and curcumin against clinical subgingival plaques that were resistant to metronidazole. The sensitivity profile of the samples to metronidazole was analyzed by the agar dilution method. Cell viability in the planktonic and biofilm phase was assessed by CFU / mL. The composition of the biofilm was evaluated by the checkboard DNA-DNA Hibrydization technique. Photosensitizers internalization was qualitatively assessed by confocal fluorescence microscopy (CLSM). The aPDT mediated by the three photosensitizers tested was able to reduce the totality of the planktonic microbial load and partially reduce the biofilm samples. The analysis performed by CLSM showed that the photosensitizers used in the application of aPDT were able to permeate the interior of the biofilm. The aPDT has been shown to be useful in a supportive and effective approach to the treatment of periodontal disease.

Topics: Anti-Bacterial Agents; Biofilms; Chlorophyllides; Curcumin; Dental Plaque; Drug Resistance, Bacterial; Humans; Methylene Blue; Metronidazole; Microbial Sensitivity Tests; Periodontal Diseases; Photochemotherapy; Photosensitizing Agents; Porphyrins

Photodynamic inactivation (PDI) may be a very promising alternative method for the antimicrobial treatment of periodontitis. Several studies have demonstrated the sensitivity of subgingival flora to PDI using toluidine blue, methylene blue, and chlorin e6 derivatives. In the present study we report the activity of the Fotolon sensitizer, composed of chlorin e6 and polyvinylpyrrolidone (PVP), against anaerobic bacteria isolated from periodontal diseases. Over 99.9% reduction in colony forming units in 20 Gram-positive and 30 Gram-negative clinical anaerobic strains was obtained.

Topics: Bacteria, Anaerobic; Chlorophyllides; Gram-Negative Anaerobic Bacteria; Gram-Positive Bacteria; Humans; In Vitro Techniques; Periodontal Diseases; Photochemotherapy; Photosensitizing Agents; Porphyrins

The aim of this study was to evaluate a new approach for killing periodontopathogenic bacteria using photodynamic therapy (PDT).. In this study, we investigated the photosensitizers chlorin e6, BLC 1010, and BLC 1014 by three different methods for their effect in PDT on the viability of periodontopathogenic bacterial species. The methods included examination of inhibition zones on agar plates, determination of colony-forming units (CFU), and the use of a bacterial viability kit.. Using the CFU method, we were able to demonstrate that the anaerobic bacteria Porphyromonas gingivalis, Fusobacterium nucleatum, and Capnocytophaga gingivalis can be photoinactivated completely by illumination with an intensity of 5.3 J/cm2 in the presence of 10 microM chlorin e6 and 10 microM BLC 1010. With the photosensitizers chlorin e6 and BLC 1010, we were able to induce zones of inhibition on agar plates. BLC 1014 failed to produce a zone of inhibition. The results of the bacterial viability test also showed that the photosensitizer BLC 1014 provides the lowest photodynamic effect in comparison to the others.. The data collected to date suggest that photodynamic therapy with chlorin e6 and BLC 1010 is advantageous for suppressing periodontopathogenic bacteria.

Topics: Aggregatibacter actinomycetemcomitans; Capnocytophaga; Chlorophyllides; Colony Count, Microbial; Eikenella corrodens; Fusobacterium nucleatum; Gram-Negative Bacteria; Humans; Lasers; Periodontal Diseases; Photochemotherapy; Photosensitizing Agents; Porphyrins; Porphyromonas gingivalis; Radiation-Sensitizing Agents

This study explores a new approach for antimicrobial therapy with light activation of targeted poly-L-lysine (pL)-chlorin e6 (ce6) conjugates. The goal was to test the hypothesis that these conjugates between pL and ce6 would efficiently target photodestruction towards gram-positive (Actinomyces viscosus) and gram-negative (Porphyromonas gingivalis) oral species while sparing an oral epithelial cell line (HCPC-1). Conjugates of ce6 with pL (average molecular weight, 2,000) having a positive, neutral, or negative charge were prepared. Illumination with red light (lambdamax = 671 nm) from a diode array produced a dose-dependent loss of CFU from the bacteria, under conditions that did not affect the viability of the epithelial cells. For P. gingivalis, the cationic conjugate produced 99% killing, while the neutral conjugate killed 91% and the anionic conjugate killed 76% after 1 min of incubation and exposure to red light for 10 min. For A. viscosus, the cationic conjugate produced >99.99% killing while HCPC-1 cells remained intact. The importance of the positive charge was shown by the effectiveness of ce6-monoethylenediamine monoamide (a monocationic derivative of ce6) in killing both bacteria. The clinically employed benzoporphyrin derivative under the same conditions killed epithelial cells while leaving P. gingivalis relatively unharmed. A mixture of ce6 with pL did not show phototoxicity comparable with that of the cationic conjugate. These results were explained by the selective uptake of the conjugates by bacteria (20- to 100-fold) compared to that by mammalian cells, while free ce6 showed much less selectivity for bacteria (5- to 20-fold). The data suggest that the cationic pL-ce6 conjugate may have an application for the photodynamic therapy of periodontal disease.

Topics: Animals; Bacteria; Cell Line; Chlorophyllides; Cricetinae; Mouth; Periodontal Diseases; Photochemotherapy; Polylysine; Porphyrins