Total biomass of multispecies-biofilm after Photodynamic Therapy mediated by nanoemulsion-phthalocyanine

Abstract

s / Photodiagnosis and Photodynamic Therapy 12 (2015) 325–375 337 Cell death of Leishmania amazonensis promastigotes induced by photodynamic therapy Debora Picanco Aureliano1,2, Jose Angelo Lauletta Lindoso2, Sandra Regina Castro Soares2, Martha Simoes Ribeiro1 1 Centro de Lasers e Aplicacoes/IPEN/CNEN/SP, Brazil 2 Laboratorio de Soroepidemiologia e Imunologia (LIM – 38)/Instituto de Medicina Tropical/USP/SP, Brazil Leishmania amazonensis is an etiological agent of American cutaneous or mucocutaneous leishmaniasis. Photodynamic therapy (PDT) is a current approach to treat leishmaniasis. In this study we investigatedmechanisms of cell death in L. amazonensis promastigotes after PDT by flow cytometry. Infective-stage of L. amazonensis metacyclic promastigotes was isolated from stationary cultures in 96 well plates, and 107 parasites were incubated with MB (50 M and 100 M) during 10min. The samples were irradiated using a 630nm-LED (P=100mW and 260mW) during 5min. Following PDT, the promastigotes of the parasites were incubated with annexin V conjugated to fluorescein isothiocyanate (FITC) and propidium iodide (PI) for 15min according to the Annexin V-FITC labeling for kit apoptosis detection (eBioscience). The reading of 5000 events independent was standard for all tests. Our results showed a similar fluorescence percentage of parasites stained when PDT was performed with 50 M MB (85.8%) or 100 M MB (82.7%) and irradiated with P=100mW. For LED power of 260mW, the fluorescence of parasites stained with Annexin V and PI was 79.4% and 72.1% (50 Mand 100 MMB, respectively). These findings indicate that PDT using MB induces apoptosis. http://dx.doi.org/10.1016/j.pdpdt.2015.07.050 Total biomass of multispecies-biofilm after Photodynamic Therapy mediated by nanoemulsion-phthalocyanine Ewerton Garcia de Oliveira Mima, Bruna Camila Rosado, Paula Volpato Sanita, Carlos Eduardo Vergani, Antonio Claudio Tedesco, Ana Claudia Pavarina Araraquara Dental School, UNESP – Univ Estadual Paulista; Photobiology & Photomedicine Research Group, Nanobiotechnology & Tissue Engineering Center, Sao Paulo University, Brazil Oral candidosis is the most common fungal infection caused by biofilm of Candida spp., especially C. albicans, but bacterial species contribute to the formation of this biofilm. This study evaluated the total biomass of multispecies-biofilm of Streptococcus mutans, Candida albicans and Candida glabrata submitted to Photodynamic Therapy (PDT) mediated by aluminum-chloride-phthalocyanine encapsulated in cationic nanoemulsion (FC-Cl-Al-NE) associated with 660nm-LED light. Standard suspensions (106–107CFU/mL) of these microorganisms were transferred to wells of microtiter plates for the multispecies-biofilm formation (48h at 37 ◦C in candle jars). The biofilm was treated for 30minutes with FC-ClAl-NE at 31.8mM. The biofilm was irradiated with a light dose of 39.3 J/cm2 (10.7mW, 21.84mW/cm2, 30min). Additional biofilms were treated only with FC-Cl-Al-NE or light, and controls consisted of untreated biofilms. Then, each sample was evaluated by measuring the total biomass using the crystal violet staining assay (absorbance at 570nm). Data were analyzed by ANOVA and Tukey ( =0.05). No significant difference (p=0.340) was verified among groups, demonstrating that PDT was not able to reduce the total biomass of the biofilm. With the parameters employed, PDT was not able to reduce the total biomass of the biofilm evaluated. http://dx.doi.org/10.1016/j.pdpdt.2015.07.051 A novel regimen for perioral dermatitis by photodynamic therapy?