Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration
ABSTRACT: Currently, bone morphogenetic protein 2 (BMP‐2) is one of the two osteoinductive growth factors used in medical devices to promote bone formation. Typically, this protein is bought from commercial houses at high rates and in small quantities that are not enough to cover clinical needs. Bec...
- Autores:
-
Mesa Restrepo, Andrea
Alzate, Juan Fernando
Patiño González, Edwin Bairon
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2021
- Institución:
- Universidad de Antioquia
- Repositorio:
- Repositorio UdeA
- Idioma:
- eng
- OAI Identifier:
- oai:bibliotecadigital.udea.edu.co:10495/19441
- Acceso en línea:
- http://hdl.handle.net/10495/19441
- Palabra clave:
- Bone Regeneration
Fosfatasa alcalina
Alkaline Phosphatase
Células C2C12
Cells C2C12
Renaturalización
Refolding protein
rhBMP‐2
http://id.nlm.nih.gov/mesh/D001861
- Rights
- openAccess
- License
- Atribución-NoComercial-CompartirIgual 2.5 Colombia
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oai:bibliotecadigital.udea.edu.co:10495/19441 |
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UDEA2 |
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Repositorio UdeA |
repository_id_str |
|
dc.title.spa.fl_str_mv |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration |
dc.title.alternative.spa.fl_str_mv |
Proteína morfogenética de hueso 2 : expresión heteróloga y potencial en la regeneración ósea |
title |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration |
spellingShingle |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration Bone Regeneration Fosfatasa alcalina Alkaline Phosphatase Células C2C12 Cells C2C12 Renaturalización Refolding protein rhBMP‐2 http://id.nlm.nih.gov/mesh/D001861 |
title_short |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration |
title_full |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration |
title_fullStr |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration |
title_full_unstemmed |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration |
title_sort |
Bone morphogenetic protein 2 : heterologous expression and potential in bone regeneration |
dc.creator.fl_str_mv |
Mesa Restrepo, Andrea Alzate, Juan Fernando Patiño González, Edwin Bairon |
dc.contributor.author.none.fl_str_mv |
Mesa Restrepo, Andrea Alzate, Juan Fernando Patiño González, Edwin Bairon |
dc.subject.mesh.none.fl_str_mv |
Bone Regeneration |
topic |
Bone Regeneration Fosfatasa alcalina Alkaline Phosphatase Células C2C12 Cells C2C12 Renaturalización Refolding protein rhBMP‐2 http://id.nlm.nih.gov/mesh/D001861 |
dc.subject.decs.none.fl_str_mv |
Fosfatasa alcalina Alkaline Phosphatase |
dc.subject.proposal.spa.fl_str_mv |
Células C2C12 Cells C2C12 Renaturalización Refolding protein rhBMP‐2 |
dc.subject.meshuri.none.fl_str_mv |
http://id.nlm.nih.gov/mesh/D001861 |
description |
ABSTRACT: Currently, bone morphogenetic protein 2 (BMP‐2) is one of the two osteoinductive growth factors used in medical devices to promote bone formation. Typically, this protein is bought from commercial houses at high rates and in small quantities that are not enough to cover clinical needs. Because of this, it has been proposed that research centers use their own heterologous expression systems to have a constant supply of BMP‐2. The aim of this study was to standardize the heterologous expression of BMP‐2 and evaluate its osteoinductive activity in vitro. Our procedure for expression and purification was based on recombinant DNA technology using the plasmid pET‐28 and IPTG as inductor. After extracting the protein from inclusion bodies, folding it and modifying it via a redox system, we observed via electrophoresis a 26 kDa dimer. We evaluated its osteoinductive activity in myoblastic C2C12 by quantifying enzymatically the activity of alkaline phosphate (ALP) and staining mineralization nodules. ALP activity is proportional to BMP‐2 concentration, increasing 90% at 3 μg/mL. These cells form calcium nodules, mineralizing 50% of the area. |
publishDate |
2021 |
dc.date.accessioned.none.fl_str_mv |
2021-04-26T16:34:48Z |
dc.date.available.none.fl_str_mv |
2021-04-26T16:34:48Z |
dc.date.issued.none.fl_str_mv |
2021 |
dc.type.spa.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.hasversion.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/ART |
dc.type.local.spa.fl_str_mv |
Artículo de investigación |
format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
status_str |
publishedVersion |
dc.identifier.issn.none.fl_str_mv |
0304-3584 |
dc.identifier.uri.none.fl_str_mv |
http://hdl.handle.net/10495/19441 |
dc.identifier.doi.none.fl_str_mv |
10.17533/udea.acbi.v43n114a01 |
dc.identifier.eissn.none.fl_str_mv |
2145-7166 |
identifier_str_mv |
0304-3584 10.17533/udea.acbi.v43n114a01 2145-7166 |
url |
http://hdl.handle.net/10495/19441 |
dc.language.iso.spa.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartofjournalabbrev.spa.fl_str_mv |
Actual. Biol. |
dc.rights.*.fl_str_mv |
Atribución-NoComercial-CompartirIgual 2.5 Colombia |
dc.rights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
dc.rights.uri.*.fl_str_mv |
http://creativecommons.org/licenses/by-nc-sa/2.5/co/ |
dc.rights.accessrights.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
dc.rights.creativecommons.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-sa/4.0/ |
rights_invalid_str_mv |
Atribución-NoComercial-CompartirIgual 2.5 Colombia http://creativecommons.org/licenses/by-nc-sa/2.5/co/ http://purl.org/coar/access_right/c_abf2 https://creativecommons.org/licenses/by-nc-sa/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.extent.spa.fl_str_mv |
10 |
dc.format.mimetype.spa.fl_str_mv |
application/pdf |
dc.publisher.spa.fl_str_mv |
Universidad de Antioquia, Facultad de Ciencias Exactas y Naturales, Instituto de Biología |
dc.publisher.group.spa.fl_str_mv |
Grupo de Biomateriales Avanzados y Medicina Regenerativa (BAMR) Grupo de Bioquímica Estructural de Macromoléculas |
dc.publisher.place.spa.fl_str_mv |
Medellín, Colombia |
institution |
Universidad de Antioquia |
bitstream.url.fl_str_mv |
http://bibliotecadigital.udea.edu.co/bitstream/10495/19441/1/Pati%c3%b1oEdwin_2021_MorphogeneticProteinRegeneration.pdf http://bibliotecadigital.udea.edu.co/bitstream/10495/19441/2/license_rdf http://bibliotecadigital.udea.edu.co/bitstream/10495/19441/3/license.txt |
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41b8baac9d5238372fad3c2c6c965a2c e2060682c9c70d4d30c83c51448f4eed 8a4605be74aa9ea9d79846c1fba20a33 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositorio Institucional Universidad de Antioquia |
repository.mail.fl_str_mv |
andres.perez@udea.edu.co |
_version_ |
1805390118431752192 |
spelling |
Mesa Restrepo, AndreaAlzate, Juan FernandoPatiño González, Edwin Bairon2021-04-26T16:34:48Z2021-04-26T16:34:48Z20210304-3584http://hdl.handle.net/10495/1944110.17533/udea.acbi.v43n114a012145-7166ABSTRACT: Currently, bone morphogenetic protein 2 (BMP‐2) is one of the two osteoinductive growth factors used in medical devices to promote bone formation. Typically, this protein is bought from commercial houses at high rates and in small quantities that are not enough to cover clinical needs. Because of this, it has been proposed that research centers use their own heterologous expression systems to have a constant supply of BMP‐2. The aim of this study was to standardize the heterologous expression of BMP‐2 and evaluate its osteoinductive activity in vitro. Our procedure for expression and purification was based on recombinant DNA technology using the plasmid pET‐28 and IPTG as inductor. After extracting the protein from inclusion bodies, folding it and modifying it via a redox system, we observed via electrophoresis a 26 kDa dimer. We evaluated its osteoinductive activity in myoblastic C2C12 by quantifying enzymatically the activity of alkaline phosphate (ALP) and staining mineralization nodules. ALP activity is proportional to BMP‐2 concentration, increasing 90% at 3 μg/mL. These cells form calcium nodules, mineralizing 50% of the area.RESUMEN: Actualmente la proteína morfogénica ósea 2 (BMP‐2) es uno de los dos factores de crecimiento más osteoinductivos usado clínicamente en dispositivos médicos que promueve la formación de hueso. Normalmente esta proteína es comprada a altos precios y en cantidades pequeñas que no alcanzan a cubrir las necesidades clínicas. Debido a esto, se ha propuesto que centros de investigación utilicen sus propios sistemas heterólogos para obtener un suministro constante. El objetivo de este trabajo es estandarizar la expresión heteróloga de la BMP‐2 y evaluar su actividad osteoinductiva in vitro. Nuestro procedimiento para la expresión y purificación se basó en la tecnología de DNA recombinante usando el plásmido pET‐28 e IPTG como inductor. Después de extraer la proteína de los cuerpos de inclusión, renaturalizarla y modificarla usando un sistema redox, se observó mediante electroforesis un dímero de 26 kDa. Se evaluó su actividad osteoinductiva en células mioblásticas C2C12 cuantificando enzimáticamente la actividad de la fosfatasa alcalina (ALP) y realizando una tinción de los nódulos de mineralización. La actividad de la fosfatasa alcalina fue proporcional a la concentración de BMP‐2, obteniendo un aumento del 90 % a concentraciones de 3 μg/mL. Estas células formaron nódulos de calcio, mineralizando un 50 % la superficie.COL0144399COL015627510application/pdfengUniversidad de Antioquia, Facultad de Ciencias Exactas y Naturales, Instituto de BiologíaGrupo de Biomateriales Avanzados y Medicina Regenerativa (BAMR)Grupo de Bioquímica Estructural de MacromoléculasMedellín, Colombiainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARTArtículo de investigaciónhttp://purl.org/coar/version/c_970fb48d4fbd8a85Atribución-NoComercial-CompartirIgual 2.5 Colombiainfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by-nc-sa/4.0/Bone RegenerationFosfatasa alcalinaAlkaline PhosphataseCélulas C2C12Cells C2C12RenaturalizaciónRefolding proteinrhBMP‐2http://id.nlm.nih.gov/mesh/D001861Bone morphogenetic protein 2 : heterologous expression and potential in bone regenerationProteína morfogenética de hueso 2 : expresión heteróloga y potencial en la regeneración óseaActual. Biol.Actualidades Biológicas43114ORIGINALPatiñoEdwin_2021_MorphogeneticProteinRegeneration.pdfPatiñoEdwin_2021_MorphogeneticProteinRegeneration.pdfArtículo de investigaciónapplication/pdf295795http://bibliotecadigital.udea.edu.co/bitstream/10495/19441/1/Pati%c3%b1oEdwin_2021_MorphogeneticProteinRegeneration.pdf41b8baac9d5238372fad3c2c6c965a2cMD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-81051http://bibliotecadigital.udea.edu.co/bitstream/10495/19441/2/license_rdfe2060682c9c70d4d30c83c51448f4eedMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://bibliotecadigital.udea.edu.co/bitstream/10495/19441/3/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5310495/19441oai:bibliotecadigital.udea.edu.co:10495/194412021-05-16 11:48:58.34Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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 |