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Previous issue date: 2015-03-30 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / A candid?ase ? uma importante manifesta??o oral em pacientes transplantados renais.
Candida spp. possuem fatores de virul?ncia que contribuem para o processo infeccioso,
incluindo a capacidade de aderir a c?lulas epiteliais e a forma??o de biofilme em superf?cies
bi?ticas e abi?ticas. O extrato obtido a partir das folhas de Eugenia uniflora [acetona: ?gua
(7:3, v/v)] tem demonstrado atividade antif?ngica contra Candida spp. Este estudo avaliou a
influ?ncia do extrato de E. uniflora na ades?o ?s C?lulas Epiteliais Bucais Humanas (CEBH)
e forma??o de biofilme de 42 cepas de Candida spp. isoladas da cavidade oral de pacientes
transplantados renais. As cepas de Candida spp. pertencentes a um banco de microorganimos
foram reativadas e reidentificadas fenotipicamente atrav?s da metodologia cl?ssica e por
m?todos moleculares (genotipagem ABC e RAPD), quando necess?rio, para a conclus?o da
identifica??o em n?vel de esp?cie. Para os ensaios de virul?ncia avaliados in vitro, as
leveduras foram cultivadas na presen?a e aus?ncia de 1000 ?g/ mL de extrato. Uma propor??o
de 10:1 c?lulas (c?lulas Candida spp. x CEBHs) foi incubada durante 1 hora a 37 ?C, 200
rpm, fixadas com formalina a 10% e o n?mero de c?lulas de Candida aderidas a 150 CEBH
foram determinadas em microsc?pio ?ptico. Os biofilmes foram formados em microplacas de
poliestireno na presen?a e aus?ncia do extrato. A quantifica??o foi realizada com colora??o
por cristal violeta a 570 nm. Todos os isolados apresentaram-se vi?veis e na reidentifica??o
fenot?pica apresentaram caracter?sticas sugestivas de cada esp?cie identificada. Duas cepas
identificadas presuntivamente como Candida dubliniensis foram confirmadas como
pertencentes a esta esp?cie atrav?s da genotipagem ABC, e as cepas identificadas como
pertencentes ao complexo Candida parapsilosis foram diferenciadas atrav?s da genotipagem
por RAPD. Candida albicans destacou-se como a esp?cie com maior potencial de ades?o ao
epit?lio bucal, enquanto que C. tropicalis apresentou expressiva forma??o de biofilme.
Detectamos que o extrato de E. uniflora foi capaz de reduzir a ades?o ao CEBH tanto para
Candida albicans como para as esp?cies de Candida n?o-Candida albicans. Por outro lado,
apenas 16 cepas de Candida spp. (36%) apresentaram a forma??o de biofilme reduzida na
presen?a do extrato. No entanto, duas cepas de C. tropicalis altamente produtoras de biofilme
tiveram uma redu??o expressiva na capacidade de forma??o de biofilme. Este estudo refor?a a
ideia de que, al?m da inibi??o do crescimento, E. uniflora pode interferir com a express?o de
alguns fatores de virul?ncia de Candida spp., e pode ser eventualmente aplicada no futuro
como um poss?vel agente antif?ngico. / Candidiasis is a major oral manifestation in kidney transplant patients. Candida spp. possess
essential virulence factors which contribute for the infectious process, including the ability to
adhere to epithelial cells and biofilm formation. The extract obtained from the leaves of
Eugenia uniflora [acetone: water (7:3, v/v)] has demonstrated antifungal activity against
Candida spp. This study evaluated the influence of the extract of E. uniflora in adhesion to
human buccal epithelial cells (HBEC) and biofilm formation of 42 strains of Candida spp.
isolated from the oral cavity of kidney transplant patients. Candida spp. strains belonging to a
culture collection were reactivated and phenotypically re-identified by classical and molecular
methods (genotyping ABC and RAPD), when necessary, to complete the identification to the
species level. For the virulence tests evaluated in vitro, yeasts were grown in the presence and
absence of 1000 ?g/mL of the extract. A ratio of 10: 1 (Candida spp. cells x HBECs) was
incubated for 1 hour at 37 ? C, 200 rpm, fixed with 10% formalin and the number of Candida
cells adhered to 150 HBEC determined by optical microscope. Biofilms were formed on
polystyrene microplates in the presence or absence of the extract. The quantification was
performed with crystal violet staining at 570 nm. All isolates were viable and exhibited
phenotypic characteristics suggestive of each species identified. Two strains presumptively
identified as Candida dubliniensis belonged to this species as determined with genotyping
ABC, while strains identified as belonging to the Candida parapsilosis species complex were
differentiated by RAPD genotyping. Candida albicans was found to be the most adherent
species to the buccal epithelia, while C. tropicalis showed remarkable biofilm formation.We
could detect that the extract of E. uniflora was able to reduce adhesion to HBEC for both
Candida albicans and non-Candida albicans Candida species. On the other hand, only 16
Candida spp. strains (36 %) showed reduced biofilm formation. However, two highly biofilm
producer strains of C. tropicalis had an expressive reduction in biofilm formation. This study
reinforces the idea that besides growth inhibition, E. uniflora may interfere with the
expression of some virulence factors of Candida spp., and may be possibly applied in the
future as a novel antifungal agent.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/20306 |
| Date | 30 March 2015 |
| Creators | Souza, Luanda Barbara Ferreira Can?rio de |
| Contributors | 25849518860, http://lattes.cnpq.br/1463249528959656, Giordani, Raquel Brandt, 00104164042, http://lattes.cnpq.br/5032750980466610, Barros, T?nia Fraga, 21929653549, http://lattes.cnpq.br/0584865026052092, Chaves, Guilherme Maranh?o |
| Publisher | Universidade Federal do Rio Grande do Norte, PROGRAMA DE P?S-GRADUA??O EM CI?NCIAS FARMACEUTICAS, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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