Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal
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 an...
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Microbiologia Fitoterapia Cromatografia em camada delgada Toxicidade CNPQ::CIENCIAS DA SAUDE |
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Microbiologia Fitoterapia Cromatografia em camada delgada Toxicidade CNPQ::CIENCIAS DA SAUDE Costa, Maria Regina Macedo Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal |
| description |
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. |
| author2 |
Lima, Kenio Costa de |
| author_facet |
Lima, Kenio Costa de Costa, Maria Regina Macedo |
| format |
doctoralThesis |
| author |
Costa, Maria Regina Macedo |
| author_sort |
Costa, Maria Regina Macedo |
| title |
Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal |
| title_short |
Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal |
| title_full |
Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal |
| title_fullStr |
Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal |
| title_full_unstemmed |
Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal |
| title_sort |
caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal |
| publisher |
Brasil |
| publishDate |
2018 |
| url |
https://repositorio.ufrn.br/jspui/handle/123456789/24929 |
| work_keys_str_mv |
AT costamariareginamacedo caracterizacaoquimicacitotoxicidadeeacaoantimicrobianadeextratosvegetaissobremicroorganismossuperinfectantesdomeioambientebucal |
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1773965838096793600 |
| spelling |
ri-123456789-249292019-05-26T06:23:38Z Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal Costa, Maria Regina Macedo Lima, Kenio Costa de Araújo, Aurigena Antunes de Lins, Ruthineia Diógenes Alves Uchoa Costa, Edja Maria Melo de Brito Pereira, Jozinete Vieira Microbiologia Fitoterapia Cromatografia em camada delgada Toxicidade CNPQ::CIENCIAS DA SAUDE 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. A presente pesquisa teve como objetivo avaliar in vitro a ação antimicrobiana dos extratos da raiz de Solanum paniculatum (jurubeba), da folha de Spondias mombin (cajá) 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 citotóxicos. O screening fitoquímico dos extratos foi investigado por reações químicas e Cromatografia em Camada Delgada (CCD). Nas análises por CCD foram utilizadas placas de sílica gel 60; AcOEt:Ácido fórmico:MeOH:Água (10:0,5:0,6:0,2, v/v/v/v), BuOH:Ácido fórmico:Água (3:1:1, v/v/v), Clorofórmio:MeOH (8:2, v/v), tolueno:acetato de etila:ácido fórmico (5:2,5:0,25, v/v/v), acetato de etila:n-propanol:ácido acético: água (4:2:2:0,6, v/v/v/v), como fases móveis; e para detecção dos metabólitos secundários, usou-se vanilina sulfúrica, solução 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 Concentração Inibitória Mínima, Concentração Inibitória Mínima de Aderência (CIMA) Cinética bactericida (CF) e fungicida (CF) dos extratos sobre Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa e Candida albicans (cepas ATCC e isolados clínicos). Ao nível de 5% de significância 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 citotóxico dos extratos foi avaliado através da viabilidade celular de fibroblastos da linhagem 3T3. As análises por CCD sugeriram a presença de ácidos fenólicos, taninos, saponinas, alcaloides, flavonoides, ácido elágico, quercetina, canferol, cumarinas e carboidratos. Todos os extratos apresentaram ação bacteriostática e fungistática. S. paniculatum apresentou desempenho médio superior ao controle positivo e estatisticamente significativo até a diluição 1:16 (31,25 mg/mL) sobre P. aeruginosa ATCC. S. mombin apresentou desempenho médio superior à clorexidina e estatisticamente significativo até a diluição 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 médio superior ao controle positivo e estatisticamente significativo até a diluição 1:4 (125 mg/mL) e 1:16 (31,25 mg/mL) sobre Enterococcus faecalis ATCC e P. aeruginosa ATCC, respectivamente. S. paniculatum apresentou ação bactericida e fungicida sobre E. faecalis (ATCC e AB) e C. albicans (ATCC e AB) e efeito antiaderente sobre todos os micro-organismos testados até a diluição 1:512 (0,97 mg/mL), exceto para S. aureus ATCC (1:8) e P. aeruginosa ATCC (1:2). S. mombin apresentou ação bactericida sobre E. faecalis (ATCC e AB), fungicida sobre C. albicans (ATCC e AB) e ação antiaderente sobre todas as bactérias testadas, com destaque para P. aeruginosa ATCC e E. faecalis (ATTC e AB) até a diluição 1:512 (0,97 mg/mL). Lycium barbarum não apresentou ação bactericida e fungicida, entretanto apresentou efeito antiaderente até a diluição 1:32 (15,65 mg/mL) sobre P. aeruginosa ATCC. Os extratos apresentaram citotoxicidade em cultura de células nas concentrações 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 são mais eficazes frente a determinados alvos do que sobre células humanas. Conclui-se que S. paniculatum, Spondias mombin e Lycium barbarum apresentaram significativa ação antimicrobiana, justificada pelos achados farmacológicos, estimulando a pesquisa de substâncias naturais bioativas, alvo-específicas, para tratamento de infecções bucais persistentes ou refratárias. 2018-03-21T15:28:03Z 2018-03-21T15:28:03Z 2016-09-28 doctoralThesis COSTA, Maria Regina Macedo. Caracterização química, citotoxicidade e ação antimicrobiana de extratos vegetais sobre micro-organismos superinfectantes do meio ambiente bucal. 2016. 152f. Tese (Doutorado em Ciências da Saúde) - Centro de Ciências da Saúde, Universidade Federal do Rio Grande do Norte, Natal, 2016. https://repositorio.ufrn.br/jspui/handle/123456789/24929 por Acesso Aberto application/pdf Brasil UFRN PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS DA SAÚDE |