Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas
Ano de defesa: | 2020 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , , |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Nove de Julho
|
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Biofotônica Aplicada às Ciências da Saúde
|
Departamento: |
Saúde
|
País: |
Brasil
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://bibliotecatede.uninove.br/handle/tede/2609 |
Resumo: | Periodontal disease is an inflammatory response to oral biofilm and conventional treatment consists of scaling and root planing. As an adjunct to this treatment, antimicrobial photodynamic therapy (aPDT) has been proposed, which consists of the use of photosensitizer (FS) and light source for the formation of reactive oxygen species (Eros). However, there are difficulties in performing aPDT clinically and it is believed that this is due to the dye dimerization. Some authors report that in a surfactant, methylene blue decreases the tendency of dimerization. In addition, other studies show that the association of urea with methylene blue can decrease this dimerization, which would increase the effectiveness of the therapy. Thus, the objective of the study was to investigate the role of methylene blue vehicles in the photodynamic inactivation of periodontopathogenic bacteria, in addition to investigating the role of urea in the inactivation of periodontopathogens and the association of urea and surfactant in the inactivation of periodontopathogens. For this, A. actinomycetemcomitans (ATCC 29523) and P. gingivalis (ATCC 33277) were grown in microaerophilia and anaerobiosis for 48 h, respectively. Then, the experiments were carried out in triplicate, with 6 groups: 1) Control, without intervention 2) Light, only irradiated 3) FS, only the dye 4) PDT1, performed aPDT with 1 min of irradiation 5) PDT3, aPDT with irradiation of 3 min and 6) PDT5, aPDT with irradiation of 5 min. Methylene blue (AM) was used as a dye in the conventional form and surfactant in the presence of SDS (sodium dodecyl sulfate), with a final concentration of 100 µM, in addition to urea (U) with a final concentration of 2 M, and irradiated with laser λ = 660nm (Photon Lase lll, DMC, São Carlos, Brazil) with power of 100mW and radiant exposure 215J / cm2. After microbial growth, colony-forming units were counted. Optical absorption spectroscopy, which is the uptake analysis, was also performed to assess the absorbance spectrum. As a result, we observed that the control, light and FS groups of A. actinomycetemcomitans showed 1x109 cfu / mL and a microbial reduction of 7 orders of magnitude was achieved after 5 min of irradiation. In the control, light and FS groups of P. gingivalis, no significant differences were observed in relation to microbial decrease, but in the groups in which aPDT was performed, there was a difference in the pattern of microbial death with 3 and 5 min of irradiation showing complete reduction in groups with AM and AM-U-SDS. When analyzing the degree of incorporation of the dye, it is observed that A. actinomycetemcomitans incorporated 20% more than P. gingivalis in the AM and U-AM solutions. Regarding absorbance, P. gingivalis showed slightly higher absorption, and between the different media the absorption spectrum was similar. In view of the information presented, it is concluded that the different methylene blue vehicles assist in the photodynamic inactivation of periodontopathogenic bacteria and increase the effectiveness of aPDT in the described patterns, and the association of urea with methylene blue increases microbial death. |
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Prates, Renato Araujohttp://lattes.cnpq.br/7664790931310514Prates, Renato Araujohttp://lattes.cnpq.br/7664790931310514Marcos, Rodrigo Labathttp://lattes.cnpq.br/2964112163504080Ana, Patricia Aparecida dahttp://lattes.cnpq.br/2372516182094204http://lattes.cnpq.br/0485490397525915Miranda, Bianca Aparecida Godoy2021-10-22T20:35:07Z2020-12-08Miranda, Bianca Aparecida Godoy. Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas. 2020. 47 f. Dissertação( Programa de Pós-Graduação em Biofotônica Aplicada às Ciências da Saúde) - Universidade Nove de Julho, São Paulo.http://bibliotecatede.uninove.br/handle/tede/2609Periodontal disease is an inflammatory response to oral biofilm and conventional treatment consists of scaling and root planing. As an adjunct to this treatment, antimicrobial photodynamic therapy (aPDT) has been proposed, which consists of the use of photosensitizer (FS) and light source for the formation of reactive oxygen species (Eros). However, there are difficulties in performing aPDT clinically and it is believed that this is due to the dye dimerization. Some authors report that in a surfactant, methylene blue decreases the tendency of dimerization. In addition, other studies show that the association of urea with methylene blue can decrease this dimerization, which would increase the effectiveness of the therapy. Thus, the objective of the study was to investigate the role of methylene blue vehicles in the photodynamic inactivation of periodontopathogenic bacteria, in addition to investigating the role of urea in the inactivation of periodontopathogens and the association of urea and surfactant in the inactivation of periodontopathogens. For this, A. actinomycetemcomitans (ATCC 29523) and P. gingivalis (ATCC 33277) were grown in microaerophilia and anaerobiosis for 48 h, respectively. Then, the experiments were carried out in triplicate, with 6 groups: 1) Control, without intervention 2) Light, only irradiated 3) FS, only the dye 4) PDT1, performed aPDT with 1 min of irradiation 5) PDT3, aPDT with irradiation of 3 min and 6) PDT5, aPDT with irradiation of 5 min. Methylene blue (AM) was used as a dye in the conventional form and surfactant in the presence of SDS (sodium dodecyl sulfate), with a final concentration of 100 µM, in addition to urea (U) with a final concentration of 2 M, and irradiated with laser λ = 660nm (Photon Lase lll, DMC, São Carlos, Brazil) with power of 100mW and radiant exposure 215J / cm2. After microbial growth, colony-forming units were counted. Optical absorption spectroscopy, which is the uptake analysis, was also performed to assess the absorbance spectrum. As a result, we observed that the control, light and FS groups of A. actinomycetemcomitans showed 1x109 cfu / mL and a microbial reduction of 7 orders of magnitude was achieved after 5 min of irradiation. In the control, light and FS groups of P. gingivalis, no significant differences were observed in relation to microbial decrease, but in the groups in which aPDT was performed, there was a difference in the pattern of microbial death with 3 and 5 min of irradiation showing complete reduction in groups with AM and AM-U-SDS. When analyzing the degree of incorporation of the dye, it is observed that A. actinomycetemcomitans incorporated 20% more than P. gingivalis in the AM and U-AM solutions. Regarding absorbance, P. gingivalis showed slightly higher absorption, and between the different media the absorption spectrum was similar. In view of the information presented, it is concluded that the different methylene blue vehicles assist in the photodynamic inactivation of periodontopathogenic bacteria and increase the effectiveness of aPDT in the described patterns, and the association of urea with methylene blue increases microbial death.A doença periodontal é uma resposta inflamatória ao biofilme oral e o tratamento convencional consiste em raspagem e alisamento radicular. Como coadjuvante a esse tratamento, tem sido proposta a terapia fotodinâmica antimicrobiana (aPDT), que consiste na utilização de fotossensibilizador (FS) e fonte de luz para a formação de espécies reativas de oxigênio (Eros). Porém, existem dificuldades em realizar a aPDT clinicamente e acredita-se que isso seja devido à dimerização do corante. Alguns autores relatam que em meio surfactante, o azul de metileno diminui a tendencia de dimerização. Além disso, outros estudos expõem que a associação da ureia ao azul de metileno pode diminuir essa dimerização, o que aumentaria a efetividade da terapia, já que a ureia aumenta a tensão superficial da água e diminui a interação entre moléculas. Assim, o objetivo do estudo foi investigar o papel dos veículos de azul de metileno na inativação fotodinâmica de bactérias periodontopatógenas, além de investigar o papel da ureia na inativação de periodontopatógenos e a associação da ureia e veículo surfactante na inativação de periodontopatógenos. Para isso, A. actinomycetemcomitans (ATCC 29523) e P. gingivalis (ATCC 33277) foram cultivados respectivamente em microaerofilia e anaerobiose por 48 h. Em seguida, os experimentos foram realizados em triplicata, com 6 grupos: 1) Controle, sem intervenção 2) Luz, apenas irradiado 3) FS, apenas o corante 4) PDT1, realizado aPDT com 1 min de irradiação 5) PDT3, aPDT com irradiação de 3 min e 6) PDT5, aPDT com irradiação de 5 min. O azul de metileno (AM) foi utilizado como corante na forma convencional e surfactante na presença de SDS (dodecil sulfato de sódio), com concentração final de 100 µM, além da ureia (U) com concentração final de 2 M, e irradiado com laser λ = 660nm (Photon Lase lll, DMC, São Carlos, Brasil) com potência de 100mW e exposição radiante 215J/cm2. Após o crescimento microbiano, as unidades formadoras de colônias foram contadas. Espectroscopia de absorção óptica, que é a análise de uptake, também foi realizada para avaliar o espectro de absorbância. Como resultado, observamos que os grupos controle, luz e FS de A. actinomycetemcomitans apresentaram 1x109 ufc/mL e uma redução microbiana de 7 ordens de magnitude foi alcançada após 5 min de irradiação. Nos grupos controle, luz e FS de P. gingivalis, não foram observadas diferenças significativas em relação à diminuição microbiana, mas nos grupos em que aPDT foi realizada, houve diferença no padrão de morte microbiana com 3 e 5 min de irradiação mostrando redução completa nos grupos com AM e AM-U-SDS. Ao analisar o grau de incorporação do corante, observa-se que A. actinomycetemcomitans incorporou 20% mais que a P. gingivalis nas soluções de AM e U-AM. Em relação à absorbância, P. gingivalis apresentou absorção ligeiramente superior, e entre os diferentes meios o espectro de absorção foi semelhante. Diante das informações apresentadas, conclui-se que os diferentes veículos de azul de metileno auxiliam na inativação fotodinâmica de bactérias periodontopatógenas e aumentam e eficácia da aPDT nos padrões descritos, e a associação da ureia com o azul de metileno aumenta a morte microbianaSubmitted by Nadir Basilio (nadirsb@uninove.br) on 2021-10-22T20:35:07Z No. of bitstreams: 1 Bianca Aparecida Godoy Miranda.pdf: 1151920 bytes, checksum: b86d2428429e6f539f22b753d1aa8b41 (MD5)Made available in DSpace on 2021-10-22T20:35:07Z (GMT). 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dc.title.por.fl_str_mv |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas |
title |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas |
spellingShingle |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas Miranda, Bianca Aparecida Godoy fototerapia dímero periodontite ureia phototherapy dimer periodontitis urea methylene blue CIENCIAS DA SAUDE |
title_short |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas |
title_full |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas |
title_fullStr |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas |
title_full_unstemmed |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas |
title_sort |
Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas |
author |
Miranda, Bianca Aparecida Godoy |
author_facet |
Miranda, Bianca Aparecida Godoy |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Prates, Renato Araujo |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/7664790931310514 |
dc.contributor.referee1.fl_str_mv |
Prates, Renato Araujo |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/7664790931310514 |
dc.contributor.referee2.fl_str_mv |
Marcos, Rodrigo Labat |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/2964112163504080 |
dc.contributor.referee3.fl_str_mv |
Ana, Patricia Aparecida da |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/2372516182094204 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/0485490397525915 |
dc.contributor.author.fl_str_mv |
Miranda, Bianca Aparecida Godoy |
contributor_str_mv |
Prates, Renato Araujo Prates, Renato Araujo Marcos, Rodrigo Labat Ana, Patricia Aparecida da |
dc.subject.por.fl_str_mv |
fototerapia dímero periodontite ureia |
topic |
fototerapia dímero periodontite ureia phototherapy dimer periodontitis urea methylene blue CIENCIAS DA SAUDE |
dc.subject.eng.fl_str_mv |
phototherapy dimer periodontitis urea methylene blue |
dc.subject.cnpq.fl_str_mv |
CIENCIAS DA SAUDE |
description |
Periodontal disease is an inflammatory response to oral biofilm and conventional treatment consists of scaling and root planing. As an adjunct to this treatment, antimicrobial photodynamic therapy (aPDT) has been proposed, which consists of the use of photosensitizer (FS) and light source for the formation of reactive oxygen species (Eros). However, there are difficulties in performing aPDT clinically and it is believed that this is due to the dye dimerization. Some authors report that in a surfactant, methylene blue decreases the tendency of dimerization. In addition, other studies show that the association of urea with methylene blue can decrease this dimerization, which would increase the effectiveness of the therapy. Thus, the objective of the study was to investigate the role of methylene blue vehicles in the photodynamic inactivation of periodontopathogenic bacteria, in addition to investigating the role of urea in the inactivation of periodontopathogens and the association of urea and surfactant in the inactivation of periodontopathogens. For this, A. actinomycetemcomitans (ATCC 29523) and P. gingivalis (ATCC 33277) were grown in microaerophilia and anaerobiosis for 48 h, respectively. Then, the experiments were carried out in triplicate, with 6 groups: 1) Control, without intervention 2) Light, only irradiated 3) FS, only the dye 4) PDT1, performed aPDT with 1 min of irradiation 5) PDT3, aPDT with irradiation of 3 min and 6) PDT5, aPDT with irradiation of 5 min. Methylene blue (AM) was used as a dye in the conventional form and surfactant in the presence of SDS (sodium dodecyl sulfate), with a final concentration of 100 µM, in addition to urea (U) with a final concentration of 2 M, and irradiated with laser λ = 660nm (Photon Lase lll, DMC, São Carlos, Brazil) with power of 100mW and radiant exposure 215J / cm2. After microbial growth, colony-forming units were counted. Optical absorption spectroscopy, which is the uptake analysis, was also performed to assess the absorbance spectrum. As a result, we observed that the control, light and FS groups of A. actinomycetemcomitans showed 1x109 cfu / mL and a microbial reduction of 7 orders of magnitude was achieved after 5 min of irradiation. In the control, light and FS groups of P. gingivalis, no significant differences were observed in relation to microbial decrease, but in the groups in which aPDT was performed, there was a difference in the pattern of microbial death with 3 and 5 min of irradiation showing complete reduction in groups with AM and AM-U-SDS. When analyzing the degree of incorporation of the dye, it is observed that A. actinomycetemcomitans incorporated 20% more than P. gingivalis in the AM and U-AM solutions. Regarding absorbance, P. gingivalis showed slightly higher absorption, and between the different media the absorption spectrum was similar. In view of the information presented, it is concluded that the different methylene blue vehicles assist in the photodynamic inactivation of periodontopathogenic bacteria and increase the effectiveness of aPDT in the described patterns, and the association of urea with methylene blue increases microbial death. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020-12-08 |
dc.date.accessioned.fl_str_mv |
2021-10-22T20:35:07Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Miranda, Bianca Aparecida Godoy. Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas. 2020. 47 f. Dissertação( Programa de Pós-Graduação em Biofotônica Aplicada às Ciências da Saúde) - Universidade Nove de Julho, São Paulo. |
dc.identifier.uri.fl_str_mv |
http://bibliotecatede.uninove.br/handle/tede/2609 |
identifier_str_mv |
Miranda, Bianca Aparecida Godoy. Azul de metileno associado com ureia na inativação fotodinâmica de bactérias periodontopatógenas. 2020. 47 f. Dissertação( Programa de Pós-Graduação em Biofotônica Aplicada às Ciências da Saúde) - Universidade Nove de Julho, São Paulo. |
url |
http://bibliotecatede.uninove.br/handle/tede/2609 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
8765449414823306929 |
dc.relation.confidence.fl_str_mv |
600 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Nove de Julho |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Biofotônica Aplicada às Ciências da Saúde |
dc.publisher.initials.fl_str_mv |
UNINOVE |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Saúde |
publisher.none.fl_str_mv |
Universidade Nove de Julho |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da Uninove instname:Universidade Nove de Julho (UNINOVE) instacron:UNINOVE |
instname_str |
Universidade Nove de Julho (UNINOVE) |
instacron_str |
UNINOVE |
institution |
UNINOVE |
reponame_str |
Biblioteca Digital de Teses e Dissertações da Uninove |
collection |
Biblioteca Digital de Teses e Dissertações da Uninove |
bitstream.url.fl_str_mv |
http://localhost:8080/tede/bitstream/tede/2609/2/Bianca+Aparecida+Godoy+Miranda.pdf http://localhost:8080/tede/bitstream/tede/2609/1/license.txt |
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b86d2428429e6f539f22b753d1aa8b41 bd3efa91386c1718a7f26a329fdcb468 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da Uninove - Universidade Nove de Julho (UNINOVE) |
repository.mail.fl_str_mv |
bibliotecatede@uninove.br||bibliotecatede@uninove.br |
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1797250764155912192 |