Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)

Detalhes bibliográficos
Ano de defesa: 2021
Autor(a) principal: Nascimento, Matheus Vieira
Orientador(a): Pontes, Karina Matthes de Freitas
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/55996
Resumo: Previous studies suggest poly(ether-ether-ketone) (PEEK) as a potential alternative to titanium implants because of its satisfactory aesthetic and mechanical characteristics. However, its polymeric nature hinders osseointegration, which could be overcome with enzymatic treatment combining laccase and catechols. This study aimed to evaluate the effects of treatment with laccase associated with catechols on wettability, cell adhesion, and antimicrobial activity of PEEK substrates. PEEK discs (15 x 2 mm) were cut and randomly distributed in treatment groups (n = 3): controls - acetate solution (Ac), laccase enzyme (Lc) and catechols (dopa, DOc dopamine DMc; and caffeic acid ACc); experimental - laccase + dopa (LDO), laccase + dopamine (LDM) and laccase + caffeic acid (AC). All substances were individually diluted in acetate solution pH 4.75 at 0.5 mg/mL at room temperature and kept in contact with PEEK substrates for 18 hours. Changes in wettability were measured by water contact angle (WCA) 0, 10, 30 minutes, 1, 2, 4 hours 1, 2, 5, 7, 14, 21, and 30 days after treatments. The cell density (cells / mm²) of osteoblast-like cells (MG-63) was measured after 24 hours of contact, with eosin-hematoxylin staining under optical microscopy. Antimicrobial activity against S. aureus (ATCC 25923) and E. coli (ATCC 11205) was performed by counting the colony-forming units (CFU/mL) after 24 hours of contact. The data were analyzed with one-way ANOVA and ANOVA of repeated measures submitted to the post-tests of Dunnet, Sidak, and Tukey, and linear trend analysis was also performed (α = 0.05). All treatments significantly decreased the WCA of the samples compared to the acetate control (Ac - 85.6º ± 0.6; p <0.0001) at all times evaluated. The groups treated with laccase+catechols had the lowest WCA, with an emphasis on LDM at T = 0min (18.5º ± 2.3). The groups treated with laccase + catechol exhibited higher cell density of MG 63 than the group treated with Lc (p <0.05), as well as with the control group Ac acetate (p <0.05). There was no statistical difference between the control and treated groups in the reduction of CFU for both S. aureus and E. coli (p> 0.05). The surface treatment with laccase associated with catechols significantly decreased the contact angle of the PEEK substrates and improved the adhesion of MG-63, although it did not show antimicrobial activity against S. aureus and E. coli.
id UFC-7_fd5e14725aa6ceff40e8ca4ad15dc4c9
oai_identifier_str oai:repositorio.ufc.br:riufc/55996
network_acronym_str UFC-7
network_name_str Repositório Institucional da Universidade Federal do Ceará (UFC)
repository_id_str
spelling Nascimento, Matheus VieiraPontes, Karina Matthes de Freitas2021-01-12T13:13:03Z2021-01-12T13:13:03Z2021NASCIMENTO, M. V. Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona). 2020. 9 f. Dissertação (Mestrado em Odontologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, Fortaleza, 2020.http://www.repositorio.ufc.br/handle/riufc/55996Previous studies suggest poly(ether-ether-ketone) (PEEK) as a potential alternative to titanium implants because of its satisfactory aesthetic and mechanical characteristics. However, its polymeric nature hinders osseointegration, which could be overcome with enzymatic treatment combining laccase and catechols. This study aimed to evaluate the effects of treatment with laccase associated with catechols on wettability, cell adhesion, and antimicrobial activity of PEEK substrates. PEEK discs (15 x 2 mm) were cut and randomly distributed in treatment groups (n = 3): controls - acetate solution (Ac), laccase enzyme (Lc) and catechols (dopa, DOc dopamine DMc; and caffeic acid ACc); experimental - laccase + dopa (LDO), laccase + dopamine (LDM) and laccase + caffeic acid (AC). All substances were individually diluted in acetate solution pH 4.75 at 0.5 mg/mL at room temperature and kept in contact with PEEK substrates for 18 hours. Changes in wettability were measured by water contact angle (WCA) 0, 10, 30 minutes, 1, 2, 4 hours 1, 2, 5, 7, 14, 21, and 30 days after treatments. The cell density (cells / mm²) of osteoblast-like cells (MG-63) was measured after 24 hours of contact, with eosin-hematoxylin staining under optical microscopy. Antimicrobial activity against S. aureus (ATCC 25923) and E. coli (ATCC 11205) was performed by counting the colony-forming units (CFU/mL) after 24 hours of contact. The data were analyzed with one-way ANOVA and ANOVA of repeated measures submitted to the post-tests of Dunnet, Sidak, and Tukey, and linear trend analysis was also performed (α = 0.05). All treatments significantly decreased the WCA of the samples compared to the acetate control (Ac - 85.6º ± 0.6; p <0.0001) at all times evaluated. The groups treated with laccase+catechols had the lowest WCA, with an emphasis on LDM at T = 0min (18.5º ± 2.3). The groups treated with laccase + catechol exhibited higher cell density of MG 63 than the group treated with Lc (p <0.05), as well as with the control group Ac acetate (p <0.05). There was no statistical difference between the control and treated groups in the reduction of CFU for both S. aureus and E. coli (p> 0.05). The surface treatment with laccase associated with catechols significantly decreased the contact angle of the PEEK substrates and improved the adhesion of MG-63, although it did not show antimicrobial activity against S. aureus and E. coli.Estudos prévios sugerem o poli(éter-éter-cetona) (PEEK) como um potencial material alternativo aos implantes de titânio devido as suas características estéticas e mecânicas satisfatórias. Contudo, sua natureza polimérica dificulta a osseointegração, sendo o tratamento enzimático com laccase e catecóis uma alternativa para melhorar algumas propriedades do material. Portanto, o objetivo desde trabalho foi avaliar os efeitos do tratamento com laccase associada a catecóis na molhabilidade, na adesão de células e na atividade antimicrobiana de substratos de PEEK. Para tal, discos de PEEK (15 x 2 mm) foram confeccionados e distribuídos aleatoriamente em grupos de tratamentos (n=3): controles - solução de acetato (Ac), enzima laccase (Lc) e catecóis (dopa, DOc; dopamina DMc; e ácido cafeico ACc); experimentais - laccase+dopa (LDO), laccase+dopamina (LDM) e laccase+ácido cafeico (AC). Todas as substâncias foram diluídas individualmente em solução de acetato pH 4,75 a 0,5 mg/mL à temperatura ambiente e mantidas em contato com os substratos de PEEK por 18 horas. Alterações na molhabilidade foram mensuradas por ângulo de contato (AC) 0, 10, 30 minutos, 1, 2, 4 horas 1, 2, 5, 7, 14, 21 e 30 dias após os tratamentos. A densidade celular (células/mm²) de osteoblast-like cells (MG-63) foi mensurada após 24h de contato, com coloração eosina-hematoxilina sob microscopia óptica. A atividade antimicrobiana contra Staphylococcus aureus (ATCC 25923) e Escherichia coli (ATCC 11205) foi executada contabilizando-se as unidades formadoras de colônia (UFC/mL) após 24 horas de contato. Os dados foram analisados com one-way ANOVA e ANOVA de medidas repetidas submetidos aos pós-testes de Dunnet, Sidak e Tukey, além da execução de análise de tendência linear (α=0,05). Todos os tratamentos diminuíram significativamente o AC das amostras em relação ao controle acetato (Ac - 85,6º ± 0,6; p<0,0001) em todos os tempos avaliados. Os grupos tratados com laccase+catecóis obtiveram os menores AC, com destaque para LDM no T=0min (18,5º ± 2,3). Os grupos tratados com laccase+catecol exibiram maior densidade celular de MG-63 que o grupo tratado com Lc (p<0,05), bem como com relação ao grupo controle acetato Ac (p<0,05). Não houve diferença estatística entre os grupos controles e os tratados na redução de UFC tanto para S. aureus quanto para E. coli (p>0,05). Conclui-se que o tratamento de superfície com laccase associado a catecóis diminuiu significativamente o ângulo de contato do substrato PEEK e melhorou a adesão de MG-63, embora não tenha apresentado atividade antimicrobiana contra S. aureus e E. coli.LacaseMolhabilidadeImplantes DentáriosCatecóisTratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/55996/4/license.txt8a4605be74aa9ea9d79846c1fba20a33MD54ORIGINAL2020_dis_mvnascimento.pdf2020_dis_mvnascimento.pdfapplication/pdf979279http://repositorio.ufc.br/bitstream/riufc/55996/3/2020_dis_mvnascimento.pdf0721347dbfe09e6fb649086380b7109bMD53riufc/559962021-01-12 10:13:03.397oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2021-01-12T13:13:03Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
title Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
spellingShingle Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
Nascimento, Matheus Vieira
Lacase
Molhabilidade
Implantes Dentários
Catecóis
title_short Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
title_full Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
title_fullStr Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
title_full_unstemmed Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
title_sort Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona)
author Nascimento, Matheus Vieira
author_facet Nascimento, Matheus Vieira
author_role author
dc.contributor.author.fl_str_mv Nascimento, Matheus Vieira
dc.contributor.advisor1.fl_str_mv Pontes, Karina Matthes de Freitas
contributor_str_mv Pontes, Karina Matthes de Freitas
dc.subject.por.fl_str_mv Lacase
Molhabilidade
Implantes Dentários
Catecóis
topic Lacase
Molhabilidade
Implantes Dentários
Catecóis
description Previous studies suggest poly(ether-ether-ketone) (PEEK) as a potential alternative to titanium implants because of its satisfactory aesthetic and mechanical characteristics. However, its polymeric nature hinders osseointegration, which could be overcome with enzymatic treatment combining laccase and catechols. This study aimed to evaluate the effects of treatment with laccase associated with catechols on wettability, cell adhesion, and antimicrobial activity of PEEK substrates. PEEK discs (15 x 2 mm) were cut and randomly distributed in treatment groups (n = 3): controls - acetate solution (Ac), laccase enzyme (Lc) and catechols (dopa, DOc dopamine DMc; and caffeic acid ACc); experimental - laccase + dopa (LDO), laccase + dopamine (LDM) and laccase + caffeic acid (AC). All substances were individually diluted in acetate solution pH 4.75 at 0.5 mg/mL at room temperature and kept in contact with PEEK substrates for 18 hours. Changes in wettability were measured by water contact angle (WCA) 0, 10, 30 minutes, 1, 2, 4 hours 1, 2, 5, 7, 14, 21, and 30 days after treatments. The cell density (cells / mm²) of osteoblast-like cells (MG-63) was measured after 24 hours of contact, with eosin-hematoxylin staining under optical microscopy. Antimicrobial activity against S. aureus (ATCC 25923) and E. coli (ATCC 11205) was performed by counting the colony-forming units (CFU/mL) after 24 hours of contact. The data were analyzed with one-way ANOVA and ANOVA of repeated measures submitted to the post-tests of Dunnet, Sidak, and Tukey, and linear trend analysis was also performed (α = 0.05). All treatments significantly decreased the WCA of the samples compared to the acetate control (Ac - 85.6º ± 0.6; p <0.0001) at all times evaluated. The groups treated with laccase+catechols had the lowest WCA, with an emphasis on LDM at T = 0min (18.5º ± 2.3). The groups treated with laccase + catechol exhibited higher cell density of MG 63 than the group treated with Lc (p <0.05), as well as with the control group Ac acetate (p <0.05). There was no statistical difference between the control and treated groups in the reduction of CFU for both S. aureus and E. coli (p> 0.05). The surface treatment with laccase associated with catechols significantly decreased the contact angle of the PEEK substrates and improved the adhesion of MG-63, although it did not show antimicrobial activity against S. aureus and E. coli.
publishDate 2021
dc.date.accessioned.fl_str_mv 2021-01-12T13:13:03Z
dc.date.available.fl_str_mv 2021-01-12T13:13:03Z
dc.date.issued.fl_str_mv 2021
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 NASCIMENTO, M. V. Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona). 2020. 9 f. Dissertação (Mestrado em Odontologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, Fortaleza, 2020.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/55996
identifier_str_mv NASCIMENTO, M. V. Tratamento enzimático com laccase associada a catecóis na molhabilidade, adesão de células e atividade antimicrobiana em substratos de poli(éter-éter-cetona). 2020. 9 f. Dissertação (Mestrado em Odontologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, Fortaleza, 2020.
url http://www.repositorio.ufc.br/handle/riufc/55996
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
bitstream.url.fl_str_mv http://repositorio.ufc.br/bitstream/riufc/55996/4/license.txt
http://repositorio.ufc.br/bitstream/riufc/55996/3/2020_dis_mvnascimento.pdf
bitstream.checksum.fl_str_mv 8a4605be74aa9ea9d79846c1fba20a33
0721347dbfe09e6fb649086380b7109b
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
_version_ 1847793239433674752