Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias

Detalhes bibliográficos
Autor(a) principal: Gusmão, Isabel Celeste Caires Pereira
Data de Publicação: 2019
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFPB
Texto Completo: https://repositorio.ufpb.br/jspui/handle/123456789/20149
Resumo: Essential oils have been considered as a promising therapeutic option in periodontal diseases. The objective of this study has been to develop pharmaceutical formulation containing Lippia sidoides Cham oil for control of periodontal diseases. The botanical material (leaves) was collected from the Herbarium of Federal University of Paraíba. The oil was extracted by steam drag and characterized by Nuclear Magnetic Resonance (NMR) and gas chromatography coupled to mass spectrometry (GC-MS). ATCC strains of Aggregatibacter actinomycetecomitans (A. actinomycetecomitans), and Porphyromonas gingivalis (P. gingivalis) were used to evaluate the antimicrobial activity of OELS and the major constituents by broth microdilution technique. The developed pharmaceutical form was orabase paste with two OELS concentrations: 0.15 and 0.25%. Prior to clinical trials, a cytotoxicity experiment and cell damage evaluation by TEM (Transmission Electron Microscopy) and SEM (Scanning Electron Microscopy) biofilm were performed. The phase I clinical trial was developed with 30 participants who used three products: placebo orabase (P, n = 10), orabase with 0.15% OELS (A, n = 10) and orabase with 0.25% OELS. (B, n = 10). The evaluations have been at time zero (TO) before use and after 6 hours of use (T6) and by following techniques: exfoliative cytology with staining, bacterial cell count in saliva and organoleptic tests. NMR and GC-MS have showed that thymol is the major component of OELS. For A. actinomycetemcomitans, the Minimum Inhibitory Concentration (MIC) of OELS, TIM, CAR and CLX was 44.23, 48.25 and 28.45 and 0.042 mg / mL, respectively. For P. gingivalis those values were: 39.30, 28.15 and 28.15 and 0.053 mg / mL. Associations with CLX have indicated synergistic effect. MET images have showed cell wall damage in both microorganisms. OELS concentrations were cytotoxic from 0.15 to 0.25%. The pharmaceutical formulation was stable at room temperature. Preliminary results of 0.15% orabase paste (A) indicate lower antimicrobial effect on T6 when compared to 0.25% paste (B). On the other hand, in T6 paste B resulted in alteration in jugal mucosa cells indicating a possible inflammatory effect. It has concluded that OELS has antimicrobial effect in vitro against periodontal disease related microorganisms and has synergistic effect with CLX. Experiments indicate that OELS has antimicrobial activity in vitro, probably related to the major component (thymol). OELS Orabase Paste has potential for clinical use. The 0.15% OELS orabase paste formulation is safe for short-term intraoral use and has in vivo antimicrobial efficacy. In addition, the observation of synergistic effect of OELS allows us to infer that it is possible to reduce OELS concentrations when in combination with CLX which may potentiate the antimicrobial effect and simultaneously reduce cytotoxicity.
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spelling Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatiasÒleos essenciaisDoenças periodontaisVerbenaceaeEssential oilsPeriodontal diseasesCNPQ::CIENCIAS BIOLOGICASEssential oils have been considered as a promising therapeutic option in periodontal diseases. The objective of this study has been to develop pharmaceutical formulation containing Lippia sidoides Cham oil for control of periodontal diseases. The botanical material (leaves) was collected from the Herbarium of Federal University of Paraíba. The oil was extracted by steam drag and characterized by Nuclear Magnetic Resonance (NMR) and gas chromatography coupled to mass spectrometry (GC-MS). ATCC strains of Aggregatibacter actinomycetecomitans (A. actinomycetecomitans), and Porphyromonas gingivalis (P. gingivalis) were used to evaluate the antimicrobial activity of OELS and the major constituents by broth microdilution technique. The developed pharmaceutical form was orabase paste with two OELS concentrations: 0.15 and 0.25%. Prior to clinical trials, a cytotoxicity experiment and cell damage evaluation by TEM (Transmission Electron Microscopy) and SEM (Scanning Electron Microscopy) biofilm were performed. The phase I clinical trial was developed with 30 participants who used three products: placebo orabase (P, n = 10), orabase with 0.15% OELS (A, n = 10) and orabase with 0.25% OELS. (B, n = 10). The evaluations have been at time zero (TO) before use and after 6 hours of use (T6) and by following techniques: exfoliative cytology with staining, bacterial cell count in saliva and organoleptic tests. NMR and GC-MS have showed that thymol is the major component of OELS. For A. actinomycetemcomitans, the Minimum Inhibitory Concentration (MIC) of OELS, TIM, CAR and CLX was 44.23, 48.25 and 28.45 and 0.042 mg / mL, respectively. For P. gingivalis those values were: 39.30, 28.15 and 28.15 and 0.053 mg / mL. Associations with CLX have indicated synergistic effect. MET images have showed cell wall damage in both microorganisms. OELS concentrations were cytotoxic from 0.15 to 0.25%. The pharmaceutical formulation was stable at room temperature. Preliminary results of 0.15% orabase paste (A) indicate lower antimicrobial effect on T6 when compared to 0.25% paste (B). On the other hand, in T6 paste B resulted in alteration in jugal mucosa cells indicating a possible inflammatory effect. It has concluded that OELS has antimicrobial effect in vitro against periodontal disease related microorganisms and has synergistic effect with CLX. Experiments indicate that OELS has antimicrobial activity in vitro, probably related to the major component (thymol). OELS Orabase Paste has potential for clinical use. The 0.15% OELS orabase paste formulation is safe for short-term intraoral use and has in vivo antimicrobial efficacy. In addition, the observation of synergistic effect of OELS allows us to infer that it is possible to reduce OELS concentrations when in combination with CLX which may potentiate the antimicrobial effect and simultaneously reduce cytotoxicity.NenhumaOs óleos essenciais têm sido considerados como uma opção terapêutica promissora nas periodontopatias. O objetivo desse estudo foi desenvolver formulação farmacêutica contendo óleo de Lippia sidoides Cham para controle de Doenças Periodontais. O material botânico (folhas) foi coletado do Herbário da Universidade Federal da Paraíba. O óleo foi extraído por arraste a vapor e caracterizado por Ressonância Magnética Nuclear (RMN) e cromatografia gasosa acoplada à espectrometria de massas (CG-EM). Cepas ATCC de Aggregatibacter actinomycetecomitans (A. actinomycetecomitans) e Porphyromonas gingivalis (P. gingivalis) foram utilizadas para avaliação da atividade antimicrobiana do OELS e dos constituintes majoritários por técnica de microdiluição em caldo. A forma farmacêutica desenvolvida foi de pasta de orabase com duas concentrações de OELS: 0,15 e 0,25%. Previamente aos ensaios clínicos, realizou-se experimento de citotoxicidade e avaliação de dano celular por MET (Microscopia Eletrônica de Transmissão) e de biofilme em MEV (Microscopia Eletrônica de Varredura). O ensaio clínico de fase I foi desenvolvido com 30 participantes que usaram três produtos: orabase placebo (P, n=10), orabase com 0,15% de OELS (A, n=10) e orabase com 0,25% de OELS (B, n=10). As avaliações foram no tempo zero (TO), antes do uso e após 6 horas de uso (T6), e pelas seguintes técnicas: citologia esfoliativa com coloração, contagem de células bacterianas na saliva e testes organolépticos. O RMN e CG-EM mostraram que o timol é o componente majoritário do OELS. Para A. actinomycetecomitans, a Concentração Inibitória Minima (CIM) do OELS, TIM, CAR e CLX foi de 44,23, 48,25 e 28,45 e 0,042 mg/mL, respectivamente. Para P. gingivalis, esses valores foram: 39,30, 28,15 e 28,15 e 0,053 mg/mL. As associações com CLX indicaram efeito sinérgico. As imagens por MET mostraram dano de parede celular em ambos os microrganismos. As concentrações de OELS se mostraram citotóxicas no estudo clínico a partir de 0,15mgm/L e 0,25mg m/L. A formulação farmacêutica se mostrou estável em temperatura ambiente. Os resultados preliminares da pasta de orabase de 0,15% (A) indicam menor efeito antimicrobiano em T6, quando comparado com a pasta de 0,25% (B). Por outro lado, em T6 a pasta B resultou em alteração nas células da mucosa jugal, indicando possível efeito inflamatório. Conclui-se que OELS tem efeito antimicrobiano in vitro frente aos microrganismos relacionados com doenças do periodonto e tem efeito sinérgico com a CLX. Os experimentos indicam que o OELS possui atividade antimicrobiana in vitro, provavelmente relacionada ao componente majoritário (timol). A pasta de orabase com OELS tem potencial para uso clínico. A formulação de pasta orabase com OELS a 0,15% é segura para uso intraoral em curta duração e possui eficácia antimicrobiana in vivo. Em adição, a observação de efeito sinérgico do OELS permite inferir que é possível reduzir as concentrações de OELS, quando em associação com CLX, o que pode potenciar o efeito antimicrobiano e, simultaneamente, reduzir a citotoxicidade.Universidade Federal da ParaíbaBrasilBiotecnologiaPrograma de Pós-Graduação em BiotecnologiaUFPBSampaio, Fabio Correiahttp://lattes.cnpq.br/7549914789004407Gusmão, Isabel Celeste Caires Pereira2021-06-01T19:22:41Z2020-08-292021-06-01T19:22:41Z2019-08-29info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttps://repositorio.ufpb.br/jspui/handle/123456789/20149porhttp://creativecommons.org/licenses/by-nd/3.0/br/info:eu-repo/semantics/embargoedAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2021-06-09T19:28:27Zoai:repositorio.ufpb.br:123456789/20149Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2021-06-09T19:28:27Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false
dc.title.none.fl_str_mv Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
title Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
spellingShingle Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
Gusmão, Isabel Celeste Caires Pereira
Òleos essenciais
Doenças periodontais
Verbenaceae
Essential oils
Periodontal diseases
CNPQ::CIENCIAS BIOLOGICAS
title_short Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
title_full Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
title_fullStr Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
title_full_unstemmed Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
title_sort Avaliação laboratorial e clínica de formulação farmacêutica contendo óleo de Lippia sidoides Cham para o controle de periodontopatias
author Gusmão, Isabel Celeste Caires Pereira
author_facet Gusmão, Isabel Celeste Caires Pereira
author_role author
dc.contributor.none.fl_str_mv Sampaio, Fabio Correia
http://lattes.cnpq.br/7549914789004407
dc.contributor.author.fl_str_mv Gusmão, Isabel Celeste Caires Pereira
dc.subject.por.fl_str_mv Òleos essenciais
Doenças periodontais
Verbenaceae
Essential oils
Periodontal diseases
CNPQ::CIENCIAS BIOLOGICAS
topic Òleos essenciais
Doenças periodontais
Verbenaceae
Essential oils
Periodontal diseases
CNPQ::CIENCIAS BIOLOGICAS
description Essential oils have been considered as a promising therapeutic option in periodontal diseases. The objective of this study has been to develop pharmaceutical formulation containing Lippia sidoides Cham oil for control of periodontal diseases. The botanical material (leaves) was collected from the Herbarium of Federal University of Paraíba. The oil was extracted by steam drag and characterized by Nuclear Magnetic Resonance (NMR) and gas chromatography coupled to mass spectrometry (GC-MS). ATCC strains of Aggregatibacter actinomycetecomitans (A. actinomycetecomitans), and Porphyromonas gingivalis (P. gingivalis) were used to evaluate the antimicrobial activity of OELS and the major constituents by broth microdilution technique. The developed pharmaceutical form was orabase paste with two OELS concentrations: 0.15 and 0.25%. Prior to clinical trials, a cytotoxicity experiment and cell damage evaluation by TEM (Transmission Electron Microscopy) and SEM (Scanning Electron Microscopy) biofilm were performed. The phase I clinical trial was developed with 30 participants who used three products: placebo orabase (P, n = 10), orabase with 0.15% OELS (A, n = 10) and orabase with 0.25% OELS. (B, n = 10). The evaluations have been at time zero (TO) before use and after 6 hours of use (T6) and by following techniques: exfoliative cytology with staining, bacterial cell count in saliva and organoleptic tests. NMR and GC-MS have showed that thymol is the major component of OELS. For A. actinomycetemcomitans, the Minimum Inhibitory Concentration (MIC) of OELS, TIM, CAR and CLX was 44.23, 48.25 and 28.45 and 0.042 mg / mL, respectively. For P. gingivalis those values were: 39.30, 28.15 and 28.15 and 0.053 mg / mL. Associations with CLX have indicated synergistic effect. MET images have showed cell wall damage in both microorganisms. OELS concentrations were cytotoxic from 0.15 to 0.25%. The pharmaceutical formulation was stable at room temperature. Preliminary results of 0.15% orabase paste (A) indicate lower antimicrobial effect on T6 when compared to 0.25% paste (B). On the other hand, in T6 paste B resulted in alteration in jugal mucosa cells indicating a possible inflammatory effect. It has concluded that OELS has antimicrobial effect in vitro against periodontal disease related microorganisms and has synergistic effect with CLX. Experiments indicate that OELS has antimicrobial activity in vitro, probably related to the major component (thymol). OELS Orabase Paste has potential for clinical use. The 0.15% OELS orabase paste formulation is safe for short-term intraoral use and has in vivo antimicrobial efficacy. In addition, the observation of synergistic effect of OELS allows us to infer that it is possible to reduce OELS concentrations when in combination with CLX which may potentiate the antimicrobial effect and simultaneously reduce cytotoxicity.
publishDate 2019
dc.date.none.fl_str_mv 2019-08-29
2020-08-29
2021-06-01T19:22:41Z
2021-06-01T19:22:41Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://repositorio.ufpb.br/jspui/handle/123456789/20149
url https://repositorio.ufpb.br/jspui/handle/123456789/20149
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nd/3.0/br/
info:eu-repo/semantics/embargoedAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nd/3.0/br/
eu_rights_str_mv embargoedAccess
dc.publisher.none.fl_str_mv Universidade Federal da Paraíba
Brasil
Biotecnologia
Programa de Pós-Graduação em Biotecnologia
UFPB
publisher.none.fl_str_mv Universidade Federal da Paraíba
Brasil
Biotecnologia
Programa de Pós-Graduação em Biotecnologia
UFPB
dc.source.none.fl_str_mv reponame:Biblioteca Digital de Teses e Dissertações da UFPB
instname:Universidade Federal da Paraíba (UFPB)
instacron:UFPB
instname_str Universidade Federal da Paraíba (UFPB)
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reponame_str Biblioteca Digital de Teses e Dissertações da UFPB
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repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)
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