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Previous issue date: 2016-09-28 / A presente pesquisa teve como objetivo avaliar in vitro a a??o antimicrobiana dos extratos da raiz de Solanum paniculatum (jurubeba), da folha de Spondias mombin (caj?) e do fruto de Lycium barbarum (gojiberry) sobre micro-organimos superinfectantes do meio ambiente bucal, bem como analisar fitoquimicamente os materiais vegetais e seus potenciais efeitos citot?xicos. O screening fitoqu?mico dos extratos foi investigado por rea??es qu?micas e Cromatografia em Camada Delgada (CCD). Nas an?lises por CCD foram utilizadas placas de s?lica gel 60; AcOEt:?cido f?rmico:MeOH:?gua (10:0,5:0,6:0,2, v/v/v/v), BuOH:?cido f?rmico:?gua (3:1:1, v/v/v), Clorof?rmio:MeOH (8:2, v/v), tolueno:acetato de etila:?cido f?rmico (5:2,5:0,25, v/v/v), acetato de etila:n-propanol:?cido ac?tico: ?gua (4:2:2:0,6, v/v/v/v), como fases m?veis; e para detec??o dos metab?litos secund?rios, usou-se vanilina sulf?rica, solu??o FeCl3 1 %, Reagente Natural A 0,5%, Reagente Natural A 0,2%/UV 365 nm, KOH a 5%(v/v), e Dragendorff. Posteriormente, foi determinada a Concentra??o Inibit?ria M?nima, Concentra??o Inibit?ria M?nima de Ader?ncia (CIMA) Cin?tica bactericida (CF) e fungicida (CF) dos extratos sobre Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa e Candida albicans (cepas ATCC e isolados cl?nicos). Ao n?vel de 5% de signific?ncia aplicou-se o teste t-Student ou de Mann-Whitney (p<0,05). Cada ensaio foi realizado em duplicata e utilizou-se como controle positivo, o digluconato de clorexidina a 0,12% e nistatina 100.000 U.I. O potencial citot?xico dos extratos foi avaliado atrav?s da viabilidade celular de fibroblastos da linhagem 3T3. As an?lises por CCD sugeriram a presen?a de ?cidos fen?licos, taninos, saponinas, alcaloides, flavonoides, ?cido el?gico, quercetina, canferol, cumarinas e carboidratos. Todos os extratos apresentaram a??o bacteriost?tica e fungist?tica. S. paniculatum apresentou desempenho m?dio superior ao controle positivo e estatisticamente significativo at? a dilui??o 1:16 (31,25 mg/mL) sobre P. aeruginosa ATCC. S. mombin apresentou desempenho m?dio superior ? clorexidina e estatisticamente significativo at? a dilui??o 1:512 (0,97 mg/ mL) e 1:256 (1,95 mg/mL) sobre E. faecalis ATCC e P. aeruginosa ATCC, respectivamente. Lycium barbarum apresentou desempenho m?dio superior ao controle positivo e estatisticamente significativo at? a dilui??o 1:4 (125 mg/mL) e 1:16 (31,25 mg/mL) sobre Enterococcus faecalis ATCC e P. aeruginosa ATCC, respectivamente. S. paniculatum apresentou a??o bactericida e fungicida sobre E. faecalis (ATCC e AB) e C. albicans (ATCC e AB) e efeito antiaderente sobre todos os micro-organismos testados at? a dilui??o 1:512 (0,97 mg/mL), exceto para S. aureus ATCC (1:8) e P. aeruginosa ATCC (1:2). S. mombin apresentou a??o bactericida sobre E. faecalis (ATCC e AB), fungicida sobre C. albicans (ATCC e AB) e a??o antiaderente sobre todas as bact?rias testadas, com destaque para P. aeruginosa ATCC e E. faecalis (ATTC e AB) at? a dilui??o 1:512 (0,97 mg/mL). Lycium barbarum n?o apresentou a??o bactericida e fungicida, entretanto apresentou efeito antiaderente at? a dilui??o 1:32 (15,65 mg/mL) sobre P. aeruginosa ATCC. Os extratos apresentaram citotoxicidade em cultura de c?lulas nas concentra??es testadas (100 mg/ mL, 31,25 mg/mL, 15,65 mg/mL, 7,81 mg/mL, 6,67 mg/mL, 1,95 mg/mL e 0,97 mg/mL) suscitando a necessidade de testes que indiquem quantas vezes os produtos s?o mais eficazes frente a determinados alvos do que sobre c?lulas humanas. Conclui-se que S. paniculatum, Spondias mombin e Lycium barbarum apresentaram significativa a??o antimicrobiana, justificada pelos achados farmacol?gicos, estimulando a pesquisa de subst?ncias naturais bioativas, alvo-espec?ficas, para tratamento de infec??es bucais persistentes ou refrat?rias. / This study aimed to evaluate the in vitro antimicrobial activity of the root extract of Solanum paniculatum (jurubeba), the leaf of Spondias mombin (hog or java plum) and the fruit of Lycium barbarum (gojiberry) on superinfecting micro-organisms in the oral environment, as well as phytochemically analyze plant material and potential cytotoxic effects. The phytochemical screening of the extracts was carried out by chemical reactions and Thin Layer Chromatography (TLC). In the analysis by TLC silica gel 60 plates were used; EtOAc: formic acid: MeOH: Water (10: 0.5: 0.6: 0.2 v / v / v / v), BuOH: formic acid: water (3: 1: 1, v / v / v ) Chloroform: MeOH (8: 2, v / v) toluene: ethyl acetate: formic acid (5: 2.5: 0.25 v / v / v) and ethyl acetate: n-propanol: acid acetic acid: water (4: 2: 2: 0.6 v / v / v / v) as mobile phase; and for detection of secondary metabolites the following were used: vanillin sulfuric acid, FeCl 3 solution of 1%, 0.5% Natural Reagent A, Reagent A 0.2% Natural UV / 365 nm KOH 5% (v / v) and Dragendorff . Subsequently, the minimum inhibitory concentration, Minimum Inhibitory Concentration of Adherence (MICA) Bactericidal Kinetics (BK) and fungicidal (FC) were determined from the extracts of Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans (ATCC and clinical isolates). The t-Student test or Mann-Whitney test was applied with 5% significance (p <0.05). Each assay was performed in duplicate and chlorhexidine digluconate at 0.12% and nystatin 100,000 I.U. were used as positive controls. The cytotoxic potential of the extracts was assessed by cell viability from fibroblasts from the 3T3 cell line. Analysis by TLC suggested the presence of phenolic acids, tannins, saponins, alkaloids, flavonoids, ellagic acid, quercetin, kaempferol, coumarins and carbohydrates. All extracts showed bacteriostatic and fungistatic action. S. paniculatum demonstrated a higher mean performance than the positive control and was statistically significant up till the dilution 1:16 (31.25 mg / mL) of P. aeruginosa ATCC. S. mombin had a mean performance superior to chlorhexidine and was statistically significant at the dilutions 1: 512 (0.97 mg / mL) and 1: 256 (1.95 mg / mL) of E. faecalis ATCC and P. aeruginosa ATCC, respectively. Lycium barbarum showed a mean performance higher than the positive control and was statistically significant at the 1:4 dilution (125 mg / mL) and 1:16 (31.25 mg/mL) of Enterococcus faecalis ATCC and P. aeruginosa ATCC, respectively. S. paniculatum presented bactericidal and fungicidal action on E. faecalis (ATCC and OE) and C. albicans (ATCC and OE) and non-stick effect on all microorganisms tested until the dilution 1: 512 (0.97 mg/mL), except for S. aureus ATCC (1:8) and P. aeruginosa ATCC (1: 2). S. mombin showed bactericidal action against E. faecalis (ATCC and OE) and C. albicans (ATCC and OE) and non-stick action on all tested bacteria, especially P. aeruginosa ATCC and E. faecalis (ATTC and OE) until the dilution 1: 512 (0.97 mg/mL). Lycium barbarum showed no bactericidal and fungicidal action, but showed non-stick effect until the 1:32 dilution (15.65 mg/mL) of P. aeruginosa ATCC. The extracts showed cytotoxicity in cell culture at the concentrations tested (100 mg / mL, 31.25 mg / mL, 15.65 mg / mL, 7.81 mg / mL, 6.67 mg / mL, 1.95 mg / mL and 0.97 mg / mL) indicating the need for tests that indicate how much more the tested products are effective against the determined targets than the human cell. It can be concluded that S. paniculatum, Spondias mombin and Lycium barbarum showed significant antimicrobial action, justified by the pharmacological findings, thus stimulating the need for further research of target-specific, bioactive natural substances in the treatment of persistent or refractory oral infections.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/24929 |
| Date | 28 September 2016 |
| Creators | Costa, Maria Regina Macedo |
| Contributors | 50337157472, Ara?jo, Aurigena Antunes de, 83806059420, Lins, Ruthineia Di?genes Alves Uchoa, 87808838420, Costa, Edja Maria Melo de Brito, 85014605400, Pereira, Jozinete Vieira, 46831576400, Lima, Kenio Costa de |
| Publisher | PROGRAMA DE P?S-GRADUA??O EM CI?NCIAS DA SA?DE, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| 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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