Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://hdl.handle.net/1822/82060 |
Resumo: | Electroactive materials allow to modulate cell-materials interactions and cell fate, leading to advanced tissue regeneration strategies. Nevertheless, their effect at the cellular level is still poorly understood. In this context, the proteome analysis of C2C12 cell differentiation cultured on piezoelectric polymer films with null average surface charge (non-poled), net positive surface charge (poled +) and net negative surface charge (poled -) has been addressed. Protein/pathway alterations for skeletal muscle development were identified comparing proteomic profiles of C2C12 cells differentiated on poly(vinylidene fluoride) with similar cells differentiated on polystyrene plate (control), using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). Only significantly expressed proteins (P<0.01, analysis of variance) were used for bioinformatic analyses. A total of 37 significantly expressed proteins were detected on the C2C12 proteome with PVDF “poled –” at 24 h, whereas on the PVDF “poled +” a total of 105 significantly expressed proteins were considered. At 5 days of differentiation, the number of significantly expressed proteins decreased to 23 and 31 in cells grown on negative and positive surface charge, respectively, being the influence of surface charge more explicit in some proteins. In both cases, proteins such as Fbn1, Hspg2, Rcn3, Tgm2, Mylpf, Anxa2, Anxa6, involved in calcium-related signaling, were highly expressed during myoblast differentiation. Furthermore, some proteins involved in muscle contraction (Acta2, Anxa2, Anxa6) were detected in the PVDF “poled +” sample. Upregulation of several proteins that enhance skeletal muscle development was detected in the PVDF “poled –” sample, including Ckm (422%), Tmem14c (384%), Serpinb6a (460%), adh7 (199%), and Car3 (171%), while for the “poled +” samples these proteins were also upregulated at a smaller magnitude (254%, 317%, 253%, 123%, 72%, respectively). Other differentially expressed proteins such as Mylpf (189%), Mybph (168%), and Mbnl1 (168%) were upregulated only in PVDF “poled –” samples, while Hba-a1 levels (581%) were increased in PVDF “poled +” sample. On the other hand, cells cultured on non-poled samples have no differences with respect to the ones culured on the control, contrarily to the poled films, with overall surface charge, demonstrating the relevance of scaffolds surface charge on cell behavior. This study demonstrates that both positive and negative overall surface charge promote the differentiation of C2C12 cells through involvement of proteins related with the contraction of the skeletal muscle cells, with a more pronounced effect with the negative charged surfaces. |
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Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysisProteomic analysisActive polymersPoly(vinylidene fluoride) filmsC2C12 myoblast cellsMyotubeScience & TechnologyElectroactive materials allow to modulate cell-materials interactions and cell fate, leading to advanced tissue regeneration strategies. Nevertheless, their effect at the cellular level is still poorly understood. In this context, the proteome analysis of C2C12 cell differentiation cultured on piezoelectric polymer films with null average surface charge (non-poled), net positive surface charge (poled +) and net negative surface charge (poled -) has been addressed. Protein/pathway alterations for skeletal muscle development were identified comparing proteomic profiles of C2C12 cells differentiated on poly(vinylidene fluoride) with similar cells differentiated on polystyrene plate (control), using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). Only significantly expressed proteins (P<0.01, analysis of variance) were used for bioinformatic analyses. A total of 37 significantly expressed proteins were detected on the C2C12 proteome with PVDF “poled –” at 24 h, whereas on the PVDF “poled +” a total of 105 significantly expressed proteins were considered. At 5 days of differentiation, the number of significantly expressed proteins decreased to 23 and 31 in cells grown on negative and positive surface charge, respectively, being the influence of surface charge more explicit in some proteins. In both cases, proteins such as Fbn1, Hspg2, Rcn3, Tgm2, Mylpf, Anxa2, Anxa6, involved in calcium-related signaling, were highly expressed during myoblast differentiation. Furthermore, some proteins involved in muscle contraction (Acta2, Anxa2, Anxa6) were detected in the PVDF “poled +” sample. Upregulation of several proteins that enhance skeletal muscle development was detected in the PVDF “poled –” sample, including Ckm (422%), Tmem14c (384%), Serpinb6a (460%), adh7 (199%), and Car3 (171%), while for the “poled +” samples these proteins were also upregulated at a smaller magnitude (254%, 317%, 253%, 123%, 72%, respectively). Other differentially expressed proteins such as Mylpf (189%), Mybph (168%), and Mbnl1 (168%) were upregulated only in PVDF “poled –” samples, while Hba-a1 levels (581%) were increased in PVDF “poled +” sample. On the other hand, cells cultured on non-poled samples have no differences with respect to the ones culured on the control, contrarily to the poled films, with overall surface charge, demonstrating the relevance of scaffolds surface charge on cell behavior. This study demonstrates that both positive and negative overall surface charge promote the differentiation of C2C12 cells through involvement of proteins related with the contraction of the skeletal muscle cells, with a more pronounced effect with the negative charged surfaces.This work has been supported by FCT-Fundacao para a Ciencia e Tecnologia (FCT) under the scope of the strategic funding of UID/FIS/04650/2020 unit, and project PTDC/BTM-MAT/28237/2017. Clarisse Ribeiro thanks the FCT for the contract under the Stimulus of Scientific Employment, Individual Support (CEECIND). 3rd Edit ion (2020.04163.CEECIND). Andreia C. Gomes acknowledges a sabbatical leave scholarship from Fundacao para a Ciencia e a Tecnologia I.P. (SFRH/BSAB/127924/2016). The authors acknowledge funding by Spanish State Research Agency (AEI) and the EuDropean Regional Development Fund (ERFD) through the project PID2019-106099RB-C43/AEI/10.13039/501100011033 and from the Basque Government Industry Departments under the ELKARTEK program. Finally, we also thank Mona Britt Hansen for expert technical help. The mass spectrometry platform was kindly donated to BH by A. P. Moller og Hustru Chastine Mc-Kinney Mollers Fond til almene Formaal.American Chemical Society (ACS)Universidade do MinhoRibeiro, Sylvie OliveiraRibeiro, ClarisseMartins, Vítor M.Honoré, BentNeves, Maria TeresaGomes, Andreia CLanceros-Méndez, S.20222025-01-01T00:00:00Z2022-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/1822/82060engRibeiro, S., Ribeiro, C., Martins, V. M., Honoré, B., Neves-Petersen, M. T., Gomes, A. C., & Lanceros-Mendez, S. (2022, May 30). Understanding Myoblast Differentiation Pathways When Cultured on Electroactive Scaffolds through Proteomic Analysis. ACS Applied Materials & Interfaces. American Chemical Society (ACS). http://doi.org/10.1021/acsami.2c034441944-824410.1021/acsami.2c0344410.1021/acsami.2c0344435635507https://pubs.acs.org/doi/10.1021/acsami.2c03444info:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-12-30T01:24:33Zoai:repositorium.sdum.uminho.pt:1822/82060Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:10:05.127387Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis |
title |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis |
spellingShingle |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis Ribeiro, Sylvie Oliveira Proteomic analysis Active polymers Poly(vinylidene fluoride) films C2C12 myoblast cells Myotube Science & Technology |
title_short |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis |
title_full |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis |
title_fullStr |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis |
title_full_unstemmed |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis |
title_sort |
Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis |
author |
Ribeiro, Sylvie Oliveira |
author_facet |
Ribeiro, Sylvie Oliveira Ribeiro, Clarisse Martins, Vítor M. Honoré, Bent Neves, Maria Teresa Gomes, Andreia C Lanceros-Méndez, S. |
author_role |
author |
author2 |
Ribeiro, Clarisse Martins, Vítor M. Honoré, Bent Neves, Maria Teresa Gomes, Andreia C Lanceros-Méndez, S. |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Ribeiro, Sylvie Oliveira Ribeiro, Clarisse Martins, Vítor M. Honoré, Bent Neves, Maria Teresa Gomes, Andreia C Lanceros-Méndez, S. |
dc.subject.por.fl_str_mv |
Proteomic analysis Active polymers Poly(vinylidene fluoride) films C2C12 myoblast cells Myotube Science & Technology |
topic |
Proteomic analysis Active polymers Poly(vinylidene fluoride) films C2C12 myoblast cells Myotube Science & Technology |
description |
Electroactive materials allow to modulate cell-materials interactions and cell fate, leading to advanced tissue regeneration strategies. Nevertheless, their effect at the cellular level is still poorly understood. In this context, the proteome analysis of C2C12 cell differentiation cultured on piezoelectric polymer films with null average surface charge (non-poled), net positive surface charge (poled +) and net negative surface charge (poled -) has been addressed. Protein/pathway alterations for skeletal muscle development were identified comparing proteomic profiles of C2C12 cells differentiated on poly(vinylidene fluoride) with similar cells differentiated on polystyrene plate (control), using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). Only significantly expressed proteins (P<0.01, analysis of variance) were used for bioinformatic analyses. A total of 37 significantly expressed proteins were detected on the C2C12 proteome with PVDF “poled –” at 24 h, whereas on the PVDF “poled +” a total of 105 significantly expressed proteins were considered. At 5 days of differentiation, the number of significantly expressed proteins decreased to 23 and 31 in cells grown on negative and positive surface charge, respectively, being the influence of surface charge more explicit in some proteins. In both cases, proteins such as Fbn1, Hspg2, Rcn3, Tgm2, Mylpf, Anxa2, Anxa6, involved in calcium-related signaling, were highly expressed during myoblast differentiation. Furthermore, some proteins involved in muscle contraction (Acta2, Anxa2, Anxa6) were detected in the PVDF “poled +” sample. Upregulation of several proteins that enhance skeletal muscle development was detected in the PVDF “poled –” sample, including Ckm (422%), Tmem14c (384%), Serpinb6a (460%), adh7 (199%), and Car3 (171%), while for the “poled +” samples these proteins were also upregulated at a smaller magnitude (254%, 317%, 253%, 123%, 72%, respectively). Other differentially expressed proteins such as Mylpf (189%), Mybph (168%), and Mbnl1 (168%) were upregulated only in PVDF “poled –” samples, while Hba-a1 levels (581%) were increased in PVDF “poled +” sample. On the other hand, cells cultured on non-poled samples have no differences with respect to the ones culured on the control, contrarily to the poled films, with overall surface charge, demonstrating the relevance of scaffolds surface charge on cell behavior. This study demonstrates that both positive and negative overall surface charge promote the differentiation of C2C12 cells through involvement of proteins related with the contraction of the skeletal muscle cells, with a more pronounced effect with the negative charged surfaces. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022 2022-01-01T00:00:00Z 2025-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://hdl.handle.net/1822/82060 |
url |
https://hdl.handle.net/1822/82060 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Ribeiro, S., Ribeiro, C., Martins, V. M., Honoré, B., Neves-Petersen, M. T., Gomes, A. C., & Lanceros-Mendez, S. (2022, May 30). Understanding Myoblast Differentiation Pathways When Cultured on Electroactive Scaffolds through Proteomic Analysis. ACS Applied Materials & Interfaces. American Chemical Society (ACS). http://doi.org/10.1021/acsami.2c03444 1944-8244 10.1021/acsami.2c03444 10.1021/acsami.2c03444 35635507 https://pubs.acs.org/doi/10.1021/acsami.2c03444 |
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info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
American Chemical Society (ACS) |
publisher.none.fl_str_mv |
American Chemical Society (ACS) |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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