Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria

Autores
Laiuppa, Juan Andrés; Santillán, Graciela Edith
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The in vitro osteogenic differentiation has been intensively studied. However, it is not yet clear precisely how osteogenesis can be optimized.Changes in extracellular Ca2þ concentration ([Ca2þ]e), as well as modulation of purinergic receptors play an important role in the regulation ofosteoblasts differentiation and bone formation. In this study, we investigated the effects of a combined treatment of ATPg-S and high [Ca2þ]e(5.35 mM) on osteogenic differentiation and function of primary cell cultures from rat calvaria. Our results indicate that ATPg-S stimulates celltransition from the G0 to S phase of cell cycle, involving the PI3K signaling pathway. Treatment with 10 or 100 mM ATPg-S and [Ca2þ]e (ATP-[Ca2þ]e) for 48 h increases cell number significantly above the control. ATPg-S treatment in osteogenic medium containing [Ca2þ]e stimulatesthe gene expression of BMP-4, BMP-5, and OPN at 16, 48, and 72 h, respectively, above control. In same conditions, treatment for 6 days with10mMUTP or 100mMUDP significantly increased the ALP activity respect to control. Cells grown in osteogenic medium showed a statisticallysignificant increase in calcium deposits at 15 and 18 days, for 10mMATPg-S treatment, and at 18 and 22 days, for [Ca2þ]e treatment, respect tocontrol but ATP-[Ca2þ]e treatment shown a significant greater mineralization at 15 days respect to ATPg-S, and at 18 days respect to bothagonists. In conclusion, we demonstrated that an osteogenic medium containing 10mM ATPg-S and 5.35mM [Ca2þ]e enhance osteogenesisand mineralization by rat primary calvarial cells cultures.
Fil: Laiuppa, Juan Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Santillán, Graciela Edith. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Materia
CALCIUM
EXTRACELLULAR ATP
OSTEOGENESIS
PURINERGIC RECEPTORS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/117328
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/117328
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat CalvariaLaiuppa, Juan AndrésSantillán, Graciela EdithCALCIUMEXTRACELLULAR ATPOSTEOGENESISPURINERGIC RECEPTORShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The in vitro osteogenic differentiation has been intensively studied. However, it is not yet clear precisely how osteogenesis can be optimized.Changes in extracellular Ca2þ concentration ([Ca2þ]e), as well as modulation of purinergic receptors play an important role in the regulation ofosteoblasts differentiation and bone formation. In this study, we investigated the effects of a combined treatment of ATPg-S and high [Ca2þ]e(5.35 mM) on osteogenic differentiation and function of primary cell cultures from rat calvaria. Our results indicate that ATPg-S stimulates celltransition from the G0 to S phase of cell cycle, involving the PI3K signaling pathway. Treatment with 10 or 100 mM ATPg-S and [Ca2þ]e (ATP-[Ca2þ]e) for 48 h increases cell number significantly above the control. ATPg-S treatment in osteogenic medium containing [Ca2þ]e stimulatesthe gene expression of BMP-4, BMP-5, and OPN at 16, 48, and 72 h, respectively, above control. In same conditions, treatment for 6 days with10mMUTP or 100mMUDP significantly increased the ALP activity respect to control. Cells grown in osteogenic medium showed a statisticallysignificant increase in calcium deposits at 15 and 18 days, for 10mMATPg-S treatment, and at 18 and 22 days, for [Ca2þ]e treatment, respect tocontrol but ATP-[Ca2þ]e treatment shown a significant greater mineralization at 15 days respect to ATPg-S, and at 18 days respect to bothagonists. In conclusion, we demonstrated that an osteogenic medium containing 10mM ATPg-S and 5.35mM [Ca2þ]e enhance osteogenesisand mineralization by rat primary calvarial cells cultures.Fil: Laiuppa, Juan Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Santillán, Graciela Edith. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaVeterinary and Human Toxicology2016-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/117328Laiuppa, Juan Andrés; Santillán, Graciela Edith; Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria; Veterinary and Human Toxicology; Journal of Cellular Biochemistry; 117; 1; 11-2016; 2658-26680730-2312CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1002/jcb.25565info:eu-repo/semantics/altIdentifier/doi/10.1002/jcb.25565info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-11-05T09:48:48Zoai:ri.conicet.gov.ar:11336/117328instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-11-05 09:48:48.987CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
title Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
spellingShingle Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
Laiuppa, Juan Andrés
CALCIUM
EXTRACELLULAR ATP
OSTEOGENESIS
PURINERGIC RECEPTORS
title_short Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
title_full Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
title_fullStr Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
title_full_unstemmed Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
title_sort Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria
dc.creator.none.fl_str_mv Laiuppa, Juan Andrés
Santillán, Graciela Edith
author Laiuppa, Juan Andrés
author_facet Laiuppa, Juan Andrés
Santillán, Graciela Edith
author_role author
author2 Santillán, Graciela Edith
author2_role author
dc.subject.none.fl_str_mv CALCIUM
EXTRACELLULAR ATP
OSTEOGENESIS
PURINERGIC RECEPTORS
topic CALCIUM
EXTRACELLULAR ATP
OSTEOGENESIS
PURINERGIC RECEPTORS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The in vitro osteogenic differentiation has been intensively studied. However, it is not yet clear precisely how osteogenesis can be optimized.Changes in extracellular Ca2þ concentration ([Ca2þ]e), as well as modulation of purinergic receptors play an important role in the regulation ofosteoblasts differentiation and bone formation. In this study, we investigated the effects of a combined treatment of ATPg-S and high [Ca2þ]e(5.35 mM) on osteogenic differentiation and function of primary cell cultures from rat calvaria. Our results indicate that ATPg-S stimulates celltransition from the G0 to S phase of cell cycle, involving the PI3K signaling pathway. Treatment with 10 or 100 mM ATPg-S and [Ca2þ]e (ATP-[Ca2þ]e) for 48 h increases cell number significantly above the control. ATPg-S treatment in osteogenic medium containing [Ca2þ]e stimulatesthe gene expression of BMP-4, BMP-5, and OPN at 16, 48, and 72 h, respectively, above control. In same conditions, treatment for 6 days with10mMUTP or 100mMUDP significantly increased the ALP activity respect to control. Cells grown in osteogenic medium showed a statisticallysignificant increase in calcium deposits at 15 and 18 days, for 10mMATPg-S treatment, and at 18 and 22 days, for [Ca2þ]e treatment, respect tocontrol but ATP-[Ca2þ]e treatment shown a significant greater mineralization at 15 days respect to ATPg-S, and at 18 days respect to bothagonists. In conclusion, we demonstrated that an osteogenic medium containing 10mM ATPg-S and 5.35mM [Ca2þ]e enhance osteogenesisand mineralization by rat primary calvarial cells cultures.
Fil: Laiuppa, Juan Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Santillán, Graciela Edith. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
description The in vitro osteogenic differentiation has been intensively studied. However, it is not yet clear precisely how osteogenesis can be optimized.Changes in extracellular Ca2þ concentration ([Ca2þ]e), as well as modulation of purinergic receptors play an important role in the regulation ofosteoblasts differentiation and bone formation. In this study, we investigated the effects of a combined treatment of ATPg-S and high [Ca2þ]e(5.35 mM) on osteogenic differentiation and function of primary cell cultures from rat calvaria. Our results indicate that ATPg-S stimulates celltransition from the G0 to S phase of cell cycle, involving the PI3K signaling pathway. Treatment with 10 or 100 mM ATPg-S and [Ca2þ]e (ATP-[Ca2þ]e) for 48 h increases cell number significantly above the control. ATPg-S treatment in osteogenic medium containing [Ca2þ]e stimulatesthe gene expression of BMP-4, BMP-5, and OPN at 16, 48, and 72 h, respectively, above control. In same conditions, treatment for 6 days with10mMUTP or 100mMUDP significantly increased the ALP activity respect to control. Cells grown in osteogenic medium showed a statisticallysignificant increase in calcium deposits at 15 and 18 days, for 10mMATPg-S treatment, and at 18 and 22 days, for [Ca2þ]e treatment, respect tocontrol but ATP-[Ca2þ]e treatment shown a significant greater mineralization at 15 days respect to ATPg-S, and at 18 days respect to bothagonists. In conclusion, we demonstrated that an osteogenic medium containing 10mM ATPg-S and 5.35mM [Ca2þ]e enhance osteogenesisand mineralization by rat primary calvarial cells cultures.
publishDate 2016
dc.date.none.fl_str_mv 2016-11
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/117328
Laiuppa, Juan Andrés; Santillán, Graciela Edith; Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria; Veterinary and Human Toxicology; Journal of Cellular Biochemistry; 117; 1; 11-2016; 2658-2668
0730-2312
CONICET Digital
CONICET
url http://hdl.handle.net/11336/117328
identifier_str_mv Laiuppa, Juan Andrés; Santillán, Graciela Edith; Effect of Combined Action of Extracellular ATP and Elevated Calcium on Osteogenic Differentiation of Primary Cultures from Rat Calvaria; Veterinary and Human Toxicology; Journal of Cellular Biochemistry; 117; 1; 11-2016; 2658-2668
0730-2312
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1002/jcb.25565
info:eu-repo/semantics/altIdentifier/doi/10.1002/jcb.25565
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Veterinary and Human Toxicology
publisher.none.fl_str_mv Veterinary and Human Toxicology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.087074

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