Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel

Detalhes bibliográficos
Autor(a) principal: Colange, Ana Lucia
Data de Publicação: 2022
Outros Autores: Oliveira, Carlos Sabino de, Domingos Neto, Benedito, Araújo, Heloisa Sobreiro Selistre de, Trovatti, Eliane, Iemma, Monica Rosas da Costa
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Research, Society and Development
Texto Completo: https://rsdjournal.org/index.php/rsd/article/view/33260
Resumo: BMP´s are signaling proteins that belong to the Transforming Growth Factor-β (TGF-β) superfamily. These proteins promote the recruitment and differentiation of mesenchymal progenitor cells into bone forming cells, the osteoblasts and increase the rate of bone formation. The carrier systems to release rhBMP-2 to the action site are based on the use of free and soluble BMP incorporated into biopolymers such as collagen, gelatin, chitosan, hyaluronic acid and silk. The fused rhBMP-2-thioredoxin could be an interesting approach for new advances in the field of carrying systems of these growth factors. The fused protein thioredoxin can be useful as a coupling agent of BMP-2 to the carrier system, binding it to the surface of the matrix and it is one of the main aims of this work. The recombinant protein rhBMP-2 was produced by IPTG induction obtaining a soluble protein without the need for refolding process. The immobilization of rhBMP-2 at the surface of the TEMPO modified cellulose nanofibrils was indicated by FTIR spectroscopy. The cellular viability tests indicated increased proliferative behavior of both, C2C12 and stem cells from rats, when seeded in presence of rhBMP2 when compared to the free rhBMP2 substrate. The calcified extracellular matrix confirmed the increased activity of the rhBMP2-cellulose substrate, indicating the success of the proposed method. The cell proliferation assays indicated the method used to immobilize rhBMP2 onto the surface of the TEMPO modified cellulose was successful. The cells growth increased when compared to the reference sample free of rhBMP2.
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spelling Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel Efecto sobre la viabilidad celular y proliferación de rhBMP-2 inmovilizada en hidrogel de celulosa modificado con TEMPOEfeito na viabilidade e proliferação celular de rhBMP-2 imobilizada em hidrogel de celulose modificado por TEMPOrhBMP2Nanofibrillas de celulosa modificadas con TEMPOInmovilizaciónInjertoProliferación celular.rhBMP2Nanofibrilas de celulose modificadas por TEMPOImobilizaçãoEnxertoProliferação celular.rhBMP2TEMPO modified cellulose nanofibrilsImmobilizationGraftingCell proliferation.BMP´s are signaling proteins that belong to the Transforming Growth Factor-β (TGF-β) superfamily. These proteins promote the recruitment and differentiation of mesenchymal progenitor cells into bone forming cells, the osteoblasts and increase the rate of bone formation. The carrier systems to release rhBMP-2 to the action site are based on the use of free and soluble BMP incorporated into biopolymers such as collagen, gelatin, chitosan, hyaluronic acid and silk. The fused rhBMP-2-thioredoxin could be an interesting approach for new advances in the field of carrying systems of these growth factors. The fused protein thioredoxin can be useful as a coupling agent of BMP-2 to the carrier system, binding it to the surface of the matrix and it is one of the main aims of this work. The recombinant protein rhBMP-2 was produced by IPTG induction obtaining a soluble protein without the need for refolding process. The immobilization of rhBMP-2 at the surface of the TEMPO modified cellulose nanofibrils was indicated by FTIR spectroscopy. The cellular viability tests indicated increased proliferative behavior of both, C2C12 and stem cells from rats, when seeded in presence of rhBMP2 when compared to the free rhBMP2 substrate. The calcified extracellular matrix confirmed the increased activity of the rhBMP2-cellulose substrate, indicating the success of the proposed method. The cell proliferation assays indicated the method used to immobilize rhBMP2 onto the surface of the TEMPO modified cellulose was successful. The cells growth increased when compared to the reference sample free of rhBMP2.Las BMP son proteínas de señalización que pertenecen a la superfamilia del factor de crecimiento transformante beta (TGF-β). Estas proteínas promueven el reclutamiento y la diferenciación de células progenitoras mesenquimales en células formadoras de hueso, osteoblastos, y aumentan la tasa de formación de hueso. Los sistemas portadores para la liberación de rhBMP-2 en el sitio de acción se basan en el uso de BMP libre y soluble incorporado en biopolímeros como colágeno, gelatina, quitosano, ácido hialurónico y seda. La rhBMP-2-tioredoxina fusionada podría ser un enfoque interesante para futuros avances en el campo de los sistemas de transporte de estos factores de crecimiento. La proteína fusionada con tiorredoxina puede ser útil como agente de acoplamiento de BMP-2 al sistema transportador, uniéndola a la superficie de la matriz y es uno de los principales objetivos de este trabajo. La proteína recombinante rhBMP-2 se produjo por inducción de IPTG obteniendo una proteína soluble sin necesidad de un proceso de replegamiento. La inmovilización de rhBMP-2 en la superficie de las nanofibrillas de celulosa modificadas con TEMPO se indicó mediante espectroscopia FTIR. Las pruebas de viabilidad celular indicaron un aumento en el comportamiento proliferativo de las células madre C2C12 y de rata, cuando se sembraron en presencia de rhBMP2 en comparación con el sustrato rhBMP2 libre. La matriz extracelular calcificada confirmó el aumento de la actividad del sustrato de celulosa rhBMP2, lo que indica el éxito del método propuesto. Los ensayos de proliferación celular indicaron que el método utilizado para inmovilizar rhBMP2 en la superficie de celulosa modificada con TEMPO fue exitoso. El crecimiento celular aumentó en comparación con la muestra de referencia sin rhBMP2.As BMP's são proteínas sinalizadoras que pertencem à superfamília Fator de Crescimento Transformador Beta (TGF-β). Essas proteínas promovem o recrutamento e a diferenciação de células progenitoras mesenquimais em células formadoras de osso, os osteoblastos e aumentam a taxa de formação óssea. Os sistemas carreadores para liberação de rhBMP-2 no local de ação são baseados no uso de BMP livre e solúvel incorporada em biopolímeros como colágeno, gelatina, quitosana, ácido hialurônico e seda. A rhBMP-2-tioredoxina fundida pode ser uma abordagem interessante para novos avanços no campo de sistemas de transporte desses fatores de crescimento. A proteína fundida tiorredoxina pode ser útil como agente de acoplamento da BMP-2 ao sistema carreador, ligando-a à superfície da matriz e é um dos principais objetivos deste trabalho. A proteína recombinante rhBMP-2 foi produzida por indução de IPTG obtendo-se uma proteína solúvel sem a necessidade de processo de redobragem. A imobilização de rhBMP-2 na superfície das nanofibrilas de celulose modificadas por TEMPO foi indicada por espectroscopia FTIR. Os testes de viabilidade celular indicaram aumento do comportamento proliferativo tanto de C2C12 quanto de células-tronco de ratos, quando semeadas na presença de rhBMP2 quando comparadas ao substrato rhBMP2 livre. A matriz extracelular calcificada confirmou o aumento da atividade do substrato rhBMP2-celulose, indicando o sucesso do método proposto. Os ensaios de proliferação celular indicaram que o método usado para imobilizar rhBMP2 na superfície da celulose modificada por TEMPO foi bem-sucedido. O crescimento das células aumentou quando comparado com a amostra de referência isenta de rhBMP2.Research, Society and Development2022-08-29info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://rsdjournal.org/index.php/rsd/article/view/3326010.33448/rsd-v11i11.33260Research, Society and Development; Vol. 11 No. 11; e471111133260Research, Society and Development; Vol. 11 Núm. 11; e471111133260Research, Society and Development; v. 11 n. 11; e4711111332602525-3409reponame:Research, Society and Developmentinstname:Universidade Federal de Itajubá (UNIFEI)instacron:UNIFEIenghttps://rsdjournal.org/index.php/rsd/article/view/33260/28618Copyright (c) 2022 Ana Lucia Colange; Carlos Sabino de Oliveira; Benedito Domingos Neto; Heloisa Sobreiro Selistre de Araújo; Eliane Trovatti; Monica Rosas da Costa Iemmahttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessColange, Ana Lucia Oliveira, Carlos Sabino de Domingos Neto, BeneditoAraújo, Heloisa Sobreiro Selistre de Trovatti, Eliane Iemma, Monica Rosas da Costa2022-09-05T13:24:46Zoai:ojs.pkp.sfu.ca:article/33260Revistahttps://rsdjournal.org/index.php/rsd/indexPUBhttps://rsdjournal.org/index.php/rsd/oairsd.articles@gmail.com2525-34092525-3409opendoar:2024-01-17T09:48:58.007290Research, Society and Development - Universidade Federal de Itajubá (UNIFEI)false
dc.title.none.fl_str_mv Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
Efecto sobre la viabilidad celular y proliferación de rhBMP-2 inmovilizada en hidrogel de celulosa modificado con TEMPO
Efeito na viabilidade e proliferação celular de rhBMP-2 imobilizada em hidrogel de celulose modificado por TEMPO
title Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
spellingShingle Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
Colange, Ana Lucia
rhBMP2
Nanofibrillas de celulosa modificadas con TEMPO
Inmovilización
Injerto
Proliferación celular.
rhBMP2
Nanofibrilas de celulose modificadas por TEMPO
Imobilização
Enxerto
Proliferação celular.
rhBMP2
TEMPO modified cellulose nanofibrils
Immobilization
Grafting
Cell proliferation.
title_short Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
title_full Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
title_fullStr Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
title_full_unstemmed Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
title_sort Effect on viability and cellular proliferation of rhBMP-2 immobilized on TEMPO modified cellulose hydrogel
author Colange, Ana Lucia
author_facet Colange, Ana Lucia
Oliveira, Carlos Sabino de
Domingos Neto, Benedito
Araújo, Heloisa Sobreiro Selistre de
Trovatti, Eliane
Iemma, Monica Rosas da Costa
author_role author
author2 Oliveira, Carlos Sabino de
Domingos Neto, Benedito
Araújo, Heloisa Sobreiro Selistre de
Trovatti, Eliane
Iemma, Monica Rosas da Costa
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Colange, Ana Lucia
Oliveira, Carlos Sabino de
Domingos Neto, Benedito
Araújo, Heloisa Sobreiro Selistre de
Trovatti, Eliane
Iemma, Monica Rosas da Costa
dc.subject.por.fl_str_mv rhBMP2
Nanofibrillas de celulosa modificadas con TEMPO
Inmovilización
Injerto
Proliferación celular.
rhBMP2
Nanofibrilas de celulose modificadas por TEMPO
Imobilização
Enxerto
Proliferação celular.
rhBMP2
TEMPO modified cellulose nanofibrils
Immobilization
Grafting
Cell proliferation.
topic rhBMP2
Nanofibrillas de celulosa modificadas con TEMPO
Inmovilización
Injerto
Proliferación celular.
rhBMP2
Nanofibrilas de celulose modificadas por TEMPO
Imobilização
Enxerto
Proliferação celular.
rhBMP2
TEMPO modified cellulose nanofibrils
Immobilization
Grafting
Cell proliferation.
description BMP´s are signaling proteins that belong to the Transforming Growth Factor-β (TGF-β) superfamily. These proteins promote the recruitment and differentiation of mesenchymal progenitor cells into bone forming cells, the osteoblasts and increase the rate of bone formation. The carrier systems to release rhBMP-2 to the action site are based on the use of free and soluble BMP incorporated into biopolymers such as collagen, gelatin, chitosan, hyaluronic acid and silk. The fused rhBMP-2-thioredoxin could be an interesting approach for new advances in the field of carrying systems of these growth factors. The fused protein thioredoxin can be useful as a coupling agent of BMP-2 to the carrier system, binding it to the surface of the matrix and it is one of the main aims of this work. The recombinant protein rhBMP-2 was produced by IPTG induction obtaining a soluble protein without the need for refolding process. The immobilization of rhBMP-2 at the surface of the TEMPO modified cellulose nanofibrils was indicated by FTIR spectroscopy. The cellular viability tests indicated increased proliferative behavior of both, C2C12 and stem cells from rats, when seeded in presence of rhBMP2 when compared to the free rhBMP2 substrate. The calcified extracellular matrix confirmed the increased activity of the rhBMP2-cellulose substrate, indicating the success of the proposed method. The cell proliferation assays indicated the method used to immobilize rhBMP2 onto the surface of the TEMPO modified cellulose was successful. The cells growth increased when compared to the reference sample free of rhBMP2.
publishDate 2022
dc.date.none.fl_str_mv 2022-08-29
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://rsdjournal.org/index.php/rsd/article/view/33260
10.33448/rsd-v11i11.33260
url https://rsdjournal.org/index.php/rsd/article/view/33260
identifier_str_mv 10.33448/rsd-v11i11.33260
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://rsdjournal.org/index.php/rsd/article/view/33260/28618
dc.rights.driver.fl_str_mv https://creativecommons.org/licenses/by/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://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 Research, Society and Development
publisher.none.fl_str_mv Research, Society and Development
dc.source.none.fl_str_mv Research, Society and Development; Vol. 11 No. 11; e471111133260
Research, Society and Development; Vol. 11 Núm. 11; e471111133260
Research, Society and Development; v. 11 n. 11; e471111133260
2525-3409
reponame:Research, Society and Development
instname:Universidade Federal de Itajubá (UNIFEI)
instacron:UNIFEI
instname_str Universidade Federal de Itajubá (UNIFEI)
instacron_str UNIFEI
institution UNIFEI
reponame_str Research, Society and Development
collection Research, Society and Development
repository.name.fl_str_mv Research, Society and Development - Universidade Federal de Itajubá (UNIFEI)
repository.mail.fl_str_mv rsd.articles@gmail.com
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