Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Tipo de documento: | Tese |
Idioma: | eng |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da USP |
Texto Completo: | https://www.teses.usp.br/teses/disponiveis/25/25149/tde-07022022-111623/ |
Resumo: | This study compared the proteomic profile of the acquired enamel pellicle (AEP) of the palatal/lingual region and unstimulated saliva of patients (1) with gastroesophageal reflux disease (GERD) and erosive tooth wear (ETW) (BEWE, score 9; GE Group); (2) with GERD without ETW (BEWE = 0; GNE group); (3) without GERD without ETW (BEWE=0, Control group) using label-free shotgun quantitative proteomic analysis (nLC-ESI-MS / MS). Since in preliminary experiments we observed increased expression of different Hb subunits in patients with GERD without ETW when compared with patients with GERD and ETW, we also evaluated, in vitro, the potential of AEP modification with different concentrations of hemoglobin (Hb) to protect enamel against initial erosion. After prophylaxis, AEP was allowed to form for 120 min. The AEP was collected from the palatal/lingual surface with filter paper soaked in 3% citric acid. After extraction, proteins were identified by nLC-ESI-MS / MS. A total of 213 proteins were identified in the AEP. The GNE group had a high number of phosphorylated and calcium-binding proteins, in addition to Spectrin beta-chain isoforms found exclusively in this group. Regarding the proteomics of saliva, when the GNE group was compared with the GE group, a remarkable increase in distinct Hb subunits was observed (up to 22 times). Other increased proteins were Serum albumin (4.5-fold) and type II cytoskeletal keratin isoforms (more than 3-fold). Proteinase inhibitors, such as Alpha-1-antitrypsin and Alpha-2-macroglobulin, were identified exclusively in the GNE group. In the in vitro study, Hb at 1.0 and 4.0 mg/mL significantly reduced initial enamel erosion after 3 days. These results indicate that many proteins may be involved in the resistance of GERD patients to the ETW, with special emphasis on hemoglobin, in addition to intracellular proteins that have the potential to bind to hydroxyapatite or participate in the formation of supramolecular aggregates. These proteins may be candidates for inclusion in dental products for protection against ETW. Data from the in vitro study indicate that changing the composition of AEP by enriching with Hb at 1.0 mg/mL might be a good strategy for protection against intrinsic erosion. |
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Biblioteca Digital de Teses e Dissertações da USP |
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spelling |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosionProteoma da película adquirida do esmalte e saliva em pacientes com doença do refluxo gastroesofágico com ou sem desgaste dentário erosivo e avaliação in vitro do potencial protetor da hemoglobina contra a erosão inicialAcquired enamel pellicleErosão dentáriaGastroesophageal refluxHemoglobinHemoglobinaPelícula adquiridaProteômicaProteomicsRefluxo gastroesofágicoTooth erosionThis study compared the proteomic profile of the acquired enamel pellicle (AEP) of the palatal/lingual region and unstimulated saliva of patients (1) with gastroesophageal reflux disease (GERD) and erosive tooth wear (ETW) (BEWE, score 9; GE Group); (2) with GERD without ETW (BEWE = 0; GNE group); (3) without GERD without ETW (BEWE=0, Control group) using label-free shotgun quantitative proteomic analysis (nLC-ESI-MS / MS). Since in preliminary experiments we observed increased expression of different Hb subunits in patients with GERD without ETW when compared with patients with GERD and ETW, we also evaluated, in vitro, the potential of AEP modification with different concentrations of hemoglobin (Hb) to protect enamel against initial erosion. After prophylaxis, AEP was allowed to form for 120 min. The AEP was collected from the palatal/lingual surface with filter paper soaked in 3% citric acid. After extraction, proteins were identified by nLC-ESI-MS / MS. A total of 213 proteins were identified in the AEP. The GNE group had a high number of phosphorylated and calcium-binding proteins, in addition to Spectrin beta-chain isoforms found exclusively in this group. Regarding the proteomics of saliva, when the GNE group was compared with the GE group, a remarkable increase in distinct Hb subunits was observed (up to 22 times). Other increased proteins were Serum albumin (4.5-fold) and type II cytoskeletal keratin isoforms (more than 3-fold). Proteinase inhibitors, such as Alpha-1-antitrypsin and Alpha-2-macroglobulin, were identified exclusively in the GNE group. In the in vitro study, Hb at 1.0 and 4.0 mg/mL significantly reduced initial enamel erosion after 3 days. These results indicate that many proteins may be involved in the resistance of GERD patients to the ETW, with special emphasis on hemoglobin, in addition to intracellular proteins that have the potential to bind to hydroxyapatite or participate in the formation of supramolecular aggregates. These proteins may be candidates for inclusion in dental products for protection against ETW. Data from the in vitro study indicate that changing the composition of AEP by enriching with Hb at 1.0 mg/mL might be a good strategy for protection against intrinsic erosion.Este estudo comparou o perfil proteômico da película adquirida do esmalte (PAE) da região palatina/lingual e saliva não estimulada de pacientes (1) com doença do refluxo gastresofágico (DRGE) e desgaste dentário erosivo (DDE) (BEWE, score 9; Grupo RE); (2) com DRGE e sem DDE (BEWE = 0; grupo RSE); c) sem DRGE e sem DDE (BEWE=0, grupo Controle) usando análise proteômica quantitativa livre de marcadores shotgun (nLC-ESI-MS / MS). Também foi avaliado, in vitro, o potencial da modificação do PAE com diferentes concentrações de hemoglobina (Hb) para proteger contra a erosão inicial do esmalte, uma vez que em experimentos preliminares observamos aumento da expressão de diferentes subunidades de Hb em pacientes com DRGE sem DDE, quando comparados com pacientes com DRGE e com DDE. A PAE foi formada 120 min após a profilaxia. A película foi coletada da superfície palatina/lingual com papel de filtro umedecido em ácido cítrico a 3%. Ao todo, foram identificadas 213 proteínas na PAE. O grupo RSE apresentou um alto número de proteínas fosforiladas e de ligação ao cálcio, além de isoformas de Spectrin beta-chain encontradas exclusivamente neste grupo. Em relação à análise proteômica da saliva, quando o grupo RSE foi comparado com o grupo RE, foi observado um aumento notável em subunidades distintas de Hb (até 22 vezes). Outras proteínas aumentadas foram Serum albumin (4,5 vezes) e isoformas de Cytoskeletal keratin II (mais de 3 vezes). Os inibidores de protease, como Alfa-1-antitripsin e Alfa-2-macroglobulin, foram identificados exclusivamente no grupo RSE. No estudo in vitro, a Hb nas concentrações de 1,0 e 4,0 mg / mL reduziu significativamente a erosão inicial do esmalte após 3 dias. Estes resultados indicam que muitas proteínas podem estar envolvidas na resistência de pacientes com DRGE à ocorrência do DDE, com especial destaque para a hemoglobina, além das proteínas intracelulares que têm potencial para se ligar à hidroxiapatita ou participar da formação de agregados supramoleculares. Dessa forma, essas proteínas podem ser candidatas a serem inclusas em produtos odontológicos para proteção contra o DDE. Os dados do estudo in vitro indicam que a alteração na composição da PAE, pelo enriquecimento com Hb a 1,0 mg/mL, pode ser uma boa estratégia para proteção contra a erosão intrínseca.Biblioteca Digitais de Teses e Dissertações da USPBuzalaf, Marilia Afonso RabeloMartini, Tatiana2021-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/25/25149/tde-07022022-111623/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2024-08-02T12:12:02Zoai:teses.usp.br:tde-07022022-111623Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212024-08-02T12:12:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion Proteoma da película adquirida do esmalte e saliva em pacientes com doença do refluxo gastroesofágico com ou sem desgaste dentário erosivo e avaliação in vitro do potencial protetor da hemoglobina contra a erosão inicial |
title |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion |
spellingShingle |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion Martini, Tatiana Acquired enamel pellicle Erosão dentária Gastroesophageal reflux Hemoglobin Hemoglobina Película adquirida Proteômica Proteomics Refluxo gastroesofágico Tooth erosion |
title_short |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion |
title_full |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion |
title_fullStr |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion |
title_full_unstemmed |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion |
title_sort |
Proteomics of acquired enamel pellicle and saliva in patients with gastroesophageal reflux disease with or without erosive tooth wear and in vitro evaluation of the protective potential of hemoglobin against initial erosion |
author |
Martini, Tatiana |
author_facet |
Martini, Tatiana |
author_role |
author |
dc.contributor.none.fl_str_mv |
Buzalaf, Marilia Afonso Rabelo |
dc.contributor.author.fl_str_mv |
Martini, Tatiana |
dc.subject.por.fl_str_mv |
Acquired enamel pellicle Erosão dentária Gastroesophageal reflux Hemoglobin Hemoglobina Película adquirida Proteômica Proteomics Refluxo gastroesofágico Tooth erosion |
topic |
Acquired enamel pellicle Erosão dentária Gastroesophageal reflux Hemoglobin Hemoglobina Película adquirida Proteômica Proteomics Refluxo gastroesofágico Tooth erosion |
description |
This study compared the proteomic profile of the acquired enamel pellicle (AEP) of the palatal/lingual region and unstimulated saliva of patients (1) with gastroesophageal reflux disease (GERD) and erosive tooth wear (ETW) (BEWE, score 9; GE Group); (2) with GERD without ETW (BEWE = 0; GNE group); (3) without GERD without ETW (BEWE=0, Control group) using label-free shotgun quantitative proteomic analysis (nLC-ESI-MS / MS). Since in preliminary experiments we observed increased expression of different Hb subunits in patients with GERD without ETW when compared with patients with GERD and ETW, we also evaluated, in vitro, the potential of AEP modification with different concentrations of hemoglobin (Hb) to protect enamel against initial erosion. After prophylaxis, AEP was allowed to form for 120 min. The AEP was collected from the palatal/lingual surface with filter paper soaked in 3% citric acid. After extraction, proteins were identified by nLC-ESI-MS / MS. A total of 213 proteins were identified in the AEP. The GNE group had a high number of phosphorylated and calcium-binding proteins, in addition to Spectrin beta-chain isoforms found exclusively in this group. Regarding the proteomics of saliva, when the GNE group was compared with the GE group, a remarkable increase in distinct Hb subunits was observed (up to 22 times). Other increased proteins were Serum albumin (4.5-fold) and type II cytoskeletal keratin isoforms (more than 3-fold). Proteinase inhibitors, such as Alpha-1-antitrypsin and Alpha-2-macroglobulin, were identified exclusively in the GNE group. In the in vitro study, Hb at 1.0 and 4.0 mg/mL significantly reduced initial enamel erosion after 3 days. These results indicate that many proteins may be involved in the resistance of GERD patients to the ETW, with special emphasis on hemoglobin, in addition to intracellular proteins that have the potential to bind to hydroxyapatite or participate in the formation of supramolecular aggregates. These proteins may be candidates for inclusion in dental products for protection against ETW. Data from the in vitro study indicate that changing the composition of AEP by enriching with Hb at 1.0 mg/mL might be a good strategy for protection against intrinsic erosion. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-12-01 |
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://www.teses.usp.br/teses/disponiveis/25/25149/tde-07022022-111623/ |
url |
https://www.teses.usp.br/teses/disponiveis/25/25149/tde-07022022-111623/ |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
|
dc.rights.driver.fl_str_mv |
Liberar o conteúdo para acesso público. info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Liberar o conteúdo para acesso público. |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.coverage.none.fl_str_mv |
|
dc.publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da USP instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Biblioteca Digital de Teses e Dissertações da USP |
collection |
Biblioteca Digital de Teses e Dissertações da USP |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br |
_version_ |
1815256775804321792 |