Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis
Autor(a) principal: | |
---|---|
Data de Publicação: | 2021 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Journal of applied oral science (Online) |
Texto Completo: | https://www.revistas.usp.br/jaos/article/view/190346 |
Resumo: | Saliva is the major contributor for the protein composition of the acquired enamel pellicle (AEP), a bacteria-free organic layer formed by the selective adsorption of salivary proteins on the surface of the enamel. However, the amount of proteins that can be recovered is even smaller under in vitro condition, due to the absence of continuous salivary flow. Objective: This study developed an in vitro AEP protocol for proteomics analysis using a new formation technique with different collection solutions. Methodology: 432 bovine enamel specimens were prepared (4x4 mm) and divided into four groups (n=108). Unstimulated saliva was provided by nine subjects. The new AEP formation technique was based on saliva resupply by a new one every 30 min within 120 minutes at 37ºC under agitation. AEP was collected using an electrode filter paper soaked in the collection solutions according with the group: 1) 3% citric acid (CA); 2) 0.5% sodium dodecyl sulfate (SDS); 3) CA followed by SDS (CA+SDS); 4) SDS followed by CA (SDS+CA). The pellicles collected were processed for analysis through LC-ESI-MS/MS technique. Results: A total of 55 proteins were identified. The total numbers of proteins identified in each group were 40, 21, 28 and 41 for the groups CA, SDS, CA+SDS and SDS+CA, respectively. Twenty-three typical AEP proteins were identified in all groups, but Mucin was only found in CA and CA+SDS, while three types of PRP were not found in the SDS group. Moreover, a typical enamel protein, Enamelin, was identified in the CA+SDS group only. Conclusion: The new technique of the in vitro AEP formation through saliva replacement was essential for a higher number of the proteins identified. In addition, considering practicality, quantity and quality of identified proteins, citric acid seems to be the best solution to be used for collection of AEP proteins. |
id |
USP-17_2403f92352e2408c0e88effef64a2795 |
---|---|
oai_identifier_str |
oai:revistas.usp.br:article/190346 |
network_acronym_str |
USP-17 |
network_name_str |
Journal of applied oral science (Online) |
repository_id_str |
|
spelling |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysisPellicleEnamelSalivaProteomicsMethodsSaliva is the major contributor for the protein composition of the acquired enamel pellicle (AEP), a bacteria-free organic layer formed by the selective adsorption of salivary proteins on the surface of the enamel. However, the amount of proteins that can be recovered is even smaller under in vitro condition, due to the absence of continuous salivary flow. Objective: This study developed an in vitro AEP protocol for proteomics analysis using a new formation technique with different collection solutions. Methodology: 432 bovine enamel specimens were prepared (4x4 mm) and divided into four groups (n=108). Unstimulated saliva was provided by nine subjects. The new AEP formation technique was based on saliva resupply by a new one every 30 min within 120 minutes at 37ºC under agitation. AEP was collected using an electrode filter paper soaked in the collection solutions according with the group: 1) 3% citric acid (CA); 2) 0.5% sodium dodecyl sulfate (SDS); 3) CA followed by SDS (CA+SDS); 4) SDS followed by CA (SDS+CA). The pellicles collected were processed for analysis through LC-ESI-MS/MS technique. Results: A total of 55 proteins were identified. The total numbers of proteins identified in each group were 40, 21, 28 and 41 for the groups CA, SDS, CA+SDS and SDS+CA, respectively. Twenty-three typical AEP proteins were identified in all groups, but Mucin was only found in CA and CA+SDS, while three types of PRP were not found in the SDS group. Moreover, a typical enamel protein, Enamelin, was identified in the CA+SDS group only. Conclusion: The new technique of the in vitro AEP formation through saliva replacement was essential for a higher number of the proteins identified. In addition, considering practicality, quantity and quality of identified proteins, citric acid seems to be the best solution to be used for collection of AEP proteins.Universidade de São Paulo. Faculdade de Odontologia de Bauru2021-09-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/jaos/article/view/19034610.1590/1678-7757-2020-0189 Journal of Applied Oral Science; Vol. 28 (2020); e2020189Journal of Applied Oral Science; Vol. 28 (2020); e2020189Journal of Applied Oral Science; v. 28 (2020); e20201891678-77651678-7757reponame:Journal of applied oral science (Online)instname:Universidade de São Paulo (USP)instacron:USPenghttps://www.revistas.usp.br/jaos/article/view/190346/175752Copyright (c) 2021 Journal of Applied Oral Sciencehttp://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessPelá, Vinícius Taioqui Ventura, Talita Mendes Oliveira Buzalaf, Marília Afonso Rabelo 2021-09-03T14:20:49Zoai:revistas.usp.br:article/190346Revistahttp://www.scielo.br/jaosPUBhttps://www.revistas.usp.br/jaos/oai||jaos@usp.br1678-77651678-7757opendoar:2021-09-03T14:20:49Journal of applied oral science (Online) - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis |
title |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis |
spellingShingle |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis Pelá, Vinícius Taioqui Pellicle Enamel Saliva Proteomics Methods |
title_short |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis |
title_full |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis |
title_fullStr |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis |
title_full_unstemmed |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis |
title_sort |
Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis |
author |
Pelá, Vinícius Taioqui |
author_facet |
Pelá, Vinícius Taioqui Ventura, Talita Mendes Oliveira Buzalaf, Marília Afonso Rabelo |
author_role |
author |
author2 |
Ventura, Talita Mendes Oliveira Buzalaf, Marília Afonso Rabelo |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Pelá, Vinícius Taioqui Ventura, Talita Mendes Oliveira Buzalaf, Marília Afonso Rabelo |
dc.subject.por.fl_str_mv |
Pellicle Enamel Saliva Proteomics Methods |
topic |
Pellicle Enamel Saliva Proteomics Methods |
description |
Saliva is the major contributor for the protein composition of the acquired enamel pellicle (AEP), a bacteria-free organic layer formed by the selective adsorption of salivary proteins on the surface of the enamel. However, the amount of proteins that can be recovered is even smaller under in vitro condition, due to the absence of continuous salivary flow. Objective: This study developed an in vitro AEP protocol for proteomics analysis using a new formation technique with different collection solutions. Methodology: 432 bovine enamel specimens were prepared (4x4 mm) and divided into four groups (n=108). Unstimulated saliva was provided by nine subjects. The new AEP formation technique was based on saliva resupply by a new one every 30 min within 120 minutes at 37ºC under agitation. AEP was collected using an electrode filter paper soaked in the collection solutions according with the group: 1) 3% citric acid (CA); 2) 0.5% sodium dodecyl sulfate (SDS); 3) CA followed by SDS (CA+SDS); 4) SDS followed by CA (SDS+CA). The pellicles collected were processed for analysis through LC-ESI-MS/MS technique. Results: A total of 55 proteins were identified. The total numbers of proteins identified in each group were 40, 21, 28 and 41 for the groups CA, SDS, CA+SDS and SDS+CA, respectively. Twenty-three typical AEP proteins were identified in all groups, but Mucin was only found in CA and CA+SDS, while three types of PRP were not found in the SDS group. Moreover, a typical enamel protein, Enamelin, was identified in the CA+SDS group only. Conclusion: The new technique of the in vitro AEP formation through saliva replacement was essential for a higher number of the proteins identified. In addition, considering practicality, quantity and quality of identified proteins, citric acid seems to be the best solution to be used for collection of AEP proteins. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-09-03 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistas.usp.br/jaos/article/view/190346 10.1590/1678-7757-2020-0189 |
url |
https://www.revistas.usp.br/jaos/article/view/190346 |
identifier_str_mv |
10.1590/1678-7757-2020-0189 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/jaos/article/view/190346/175752 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2021 Journal of Applied Oral Science http://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2021 Journal of Applied Oral Science http://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Odontologia de Bauru |
publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Odontologia de Bauru |
dc.source.none.fl_str_mv |
Journal of Applied Oral Science; Vol. 28 (2020); e2020189 Journal of Applied Oral Science; Vol. 28 (2020); e2020189 Journal of Applied Oral Science; v. 28 (2020); e2020189 1678-7765 1678-7757 reponame:Journal of applied oral science (Online) instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Journal of applied oral science (Online) |
collection |
Journal of applied oral science (Online) |
repository.name.fl_str_mv |
Journal of applied oral science (Online) - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
||jaos@usp.br |
_version_ |
1800221681986306048 |