ABSTRACT
Despite many advances, biomaterial-associated infections continue to be a major clinical problem. In order to minimize bacterial adhesion, material surface modifications are currently being investigated and natural products possess large potential for the design of innovative surface coatings. We report the bioguided phytochemical investigation of Pityrocarpa moniliformis and the characterization of tannins by mass spectrometry. It was demonstrated that B-type linked proanthocyanidins-coated surfaces, here termed Green coatings, reduced Gram-positive bacterial adhesion and supported mammalian cell spreading. The proposed mechanism of bacterial attachment inhibition is based on electrostatic repulsion, high hydrophilicity and the steric hindrance provided by the coating that blocks bacterium-substratum interactions. This work shows the applicability of a prototype Green-coated surface that aims to promote necessary mammalian tissue compatibility, while reducing bacterial colonization.
Subject(s)
Bacterial Adhesion/physiology , Environmental Microbiology , Surface Properties , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Bacterial Adhesion/drug effects , Biofilms/drug effects , Biological Products/chemistry , Biological Products/pharmacology , Coated Materials, Biocompatible/chemistry , Epithelial Cells/drug effects , Fabaceae/chemistry , Gram-Positive Bacteria/drug effects , Gram-Positive Bacteria/physiology , Plant Extracts/chemistry , Plant Extracts/pharmacology , Proanthocyanidins/chemistry , Proanthocyanidins/pharmacologyABSTRACT
Giardia lamblia é um protozoário parasito do intestino delgado, agente etiológico da giardiose, o qual apresenta um complexo citoesqueleto composto por microtúbulos. Neste estudo, a distribuição dos microtúbulos no citoesqueleto de trofozoítos e cistos de G. lamblia foi investigada por meio de um taxóide fluorescente, o FLUTAX-2 - um derivado do Taxol que se liga aos dímeros polimerizados de alfabeta-tubulina. O efeito do metronidazol sobre o citoesqueleto do organismo também foi avaliado. FLUTAX-2 foi capaz de se ligar aos microtúbulos de trofozoítos e cistos de G. lamblia, tornando possível a visualização de estruturas do citoesqueleto do microrganismo, como flagelos, funis e disco adesivo. Além disso, FLUTAX-2 marcou os trofozoitos cultivados por diferentes períodos, revelando que o intervalo de 48 horas é o melhor tempo de cultivo. A incubação dos parasitos na presença de metronidazol não demonstrou alteração significativa na marcação microtubular por FLUTAX-2. Nosso grupo já apresentou a marcação pelo FLUTAX-2 em outros protozoários, Trichomonas vaginalis, T. gallinae e Tritrichomonas foetus, e este é o primeiro trabalho a revelar o perfil de ligação em G. lamblia. Os resultados contribuem para o conhecimento de aspectos biológicos e morfológicos do parasito e sugerem perspectivas para o diagnóstico laboratorial da giardiose.
G. lamblia is a protozoan parasite of the small intestine and the etiologic agent ofgiardiosis, which presents a complex cytoskeleton composed of microtubules. In this study, the microtubule cytoskeleton was labeled in trophozoites and cysts of G. lamblia, using a fluorescent taxoid (FLUTAX-2), a derivative of Taxol, which binds to áâ-tubulin dimer polymerized. In addition, the effect of metronidazole on the cytoskeleton of the parasite was also evaluated. FLUTAX-2 was able to labelthe microtubules of trophozoites and cysts, allowing the observation of cytoskeletal structures, such as flagella, funis, adhesive disk, and median body. Moreover, FLUTAX-2 labeled the trophozoites from several different cultivation times, revealing the 48 hours as the best incubation period. The incubation of parasites in presence of metronidazole did not show significant alteration on the microtubulelabeling by FLUTAX-2. We have shown the FLUTAX-2 labeling in other protozoa,Trichomonas vaginalis, T. gallinae and Tritrichomonas foetus, and this is the first report revealing the binding profile in G. lamblia. Our results contribute to the knowledge of biological and morphological features of this intestinal pathogen. Furthermore, our data showed no modification of microtubule profile labeling in presence of metronidazole. Finally, a new perspective for giardiosis diagnosticis suggested, since trophozoites and cysts of G. lamblia were labeled withFLUTAX-2.