Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto
Antecedentes: Estudios acerca de ingeniería tisular establecen que los co-cultivos directos han sido útiles para comprender la interacción entre células debido a que la comunicación intercelular promueve el intercambio de señales y factores de crecimiento que favorecen la diferenciación hacia distin...
- Autores:
-
Calderón García, Eliana
Ramírez Robles, Luz Eliana
- Tipo de recurso:
- https://purl.org/coar/resource_type/c_7a1f
- Fecha de publicación:
- 2025
- Institución:
- Universidad El Bosque
- Repositorio:
- Repositorio U. El Bosque
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unbosque.edu.co:20.500.12495/15703
- Acceso en línea:
- https://hdl.handle.net/20.500.12495/15703
- Palabra clave:
- Preosteoblastos
Células madre
Pulpa dental
Osteoblastos
Técnicas de cultivo
Diferenciación celular
Pre-osteoblasts
Stem cells
Dental Pulp
Osteoblasts
Culture techniques
Cell differentiation
WU400
- Rights
- License
- Attribution-NonCommercial-ShareAlike 4.0 International
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Repositorio U. El Bosque |
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| dc.title.none.fl_str_mv |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto |
| dc.title.translated.none.fl_str_mv |
Evaluation of the differentiation of mesenchymal cells and pre-osteoblasts in a direct and indirect co-culture system |
| title |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto |
| spellingShingle |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto Preosteoblastos Células madre Pulpa dental Osteoblastos Técnicas de cultivo Diferenciación celular Pre-osteoblasts Stem cells Dental Pulp Osteoblasts Culture techniques Cell differentiation WU400 |
| title_short |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto |
| title_full |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto |
| title_fullStr |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto |
| title_full_unstemmed |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto |
| title_sort |
Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirecto |
| dc.creator.fl_str_mv |
Calderón García, Eliana Ramírez Robles, Luz Eliana |
| dc.contributor.advisor.none.fl_str_mv |
Escobar Márquez, Lina María Bendahan Álvarez, Zita Carolina Castaño Duque, Sandra Patricia |
| dc.contributor.author.none.fl_str_mv |
Calderón García, Eliana Ramírez Robles, Luz Eliana |
| dc.subject.none.fl_str_mv |
Preosteoblastos Células madre Pulpa dental Osteoblastos Técnicas de cultivo Diferenciación celular |
| topic |
Preosteoblastos Células madre Pulpa dental Osteoblastos Técnicas de cultivo Diferenciación celular Pre-osteoblasts Stem cells Dental Pulp Osteoblasts Culture techniques Cell differentiation WU400 |
| dc.subject.keywords.none.fl_str_mv |
Pre-osteoblasts Stem cells Dental Pulp Osteoblasts Culture techniques Cell differentiation |
| dc.subject.nlm.none.fl_str_mv |
WU400 |
| description |
Antecedentes: Estudios acerca de ingeniería tisular establecen que los co-cultivos directos han sido útiles para comprender la interacción entre células debido a que la comunicación intercelular promueve el intercambio de señales y factores de crecimiento que favorecen la diferenciación hacia distintos linajes celulares. Adicionalmente, en otros estudios se reportan interacciones celulares en co-cultivos indirectos que proporcionan un panorama favorable para la generación de tejidos o la reconstrucción de defectos óseos. No obstante, no existe evidencia clara que compare los co-cultivos directos y los indirectos en el desarrollo de procesos de proliferación y diferenciación de células madre a OBs. Esto podría en un futuro establecer nuevas terapéuticas que brindan soluciones a la reparación de tejidos de diferentes patologías. Objetivo: Determinar y comparar el efecto en la diferenciación, proliferación y morfología de células madre mesenquimales (MSCs) derivadas de la pulpa dental (hDPSC) y pre-osteoblastos (Saos-2) co-cultivados de forma directa e indirecta in vitro. Materiales y Métodos: Se evaluó proliferación, viabilidad, morfología y diferenciación celular a partir de co- cultivo directo e indirecto de células de la pulpa dental (hDPSC) y Saos-2 en medio DMEM a los 7 , 14 y 21 días. Se utilizó control positivo con medio de diferenciación y control negativo con medio DMEM. Resultados: Se realizó inmunofenotipificación a las hDPSC donde se encontró células positivas para marcadores de superficie CD105, CD90, CD73 y se evaluaron características morfológicas donde se observó células fibroblastoides adheridas a la caja. El co-cultivo indirecto y el directo de células Saos-2+hDPSC, produjo un incremento significativo (P < 0.05) en el número de células y proliferación celular desde el 6to día en el co-cultivo indirecto y desde el cuarto día en el co-cultivo directo. En las células hDPSC y Saos-2 sin co-cultivo (control) no se observó tinción con el rojo de alizarina, mientras que las células tratadas con MDO como control positivo de diferenciación osteoblástica mostraron igualmente formación de nódulos de calcio. La mayor cantidad de rojo de alizarina como medida de absorbancia, se evidenció en las células co-cultivadas de forma indirecta y en las células directamente co-cultivadas . Estas absorbancias fueron significativamente mayores que las obtenidas en las células hDPSC y Saos-2 sin co-cultivo. Al analizar los cambios en la expresión de genes de diferenciación osteoblástica como RUNX2 y osteocalcina (OCN) se encontró que a los 7 días RUNX2 aumentó su expresión en todos los grupos experimentales excepto en las células hDPSC co-cultivadas de forma indirecta con Saos-2. Por otro lado, El mayor incremento en la expresión se evidenció en el co-cultivo directo (116 veces más que el control) y hDPSC tratadas con MDO (83 veces más que el control). La expresión se redujo significativamente en todos los grupos experimentales a los 21 días. Conclusiones: Este estudio sugiere que el co-cultivo con células Saos-2 y hDPSC tanto directo como indirecto es efectivo para inducir cambios morfológicos, en la proliferación y diferenciación osteoblástica. Por este motivo, pueden ser una herramienta útil en la investigación de terapias de regeneración para la reparación de defectos óseos. |
| publishDate |
2025 |
| dc.date.accessioned.none.fl_str_mv |
2025-08-12T14:15:03Z |
| dc.date.available.none.fl_str_mv |
2025-08-12T14:15:03Z |
| dc.date.issued.none.fl_str_mv |
2025-08 |
| dc.type.coar.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
| dc.type.local.spa.fl_str_mv |
Tesis/Trabajo de grado - Monografía - Especialización |
| dc.type.coar.none.fl_str_mv |
https://purl.org/coar/resource_type/c_7a1f |
| dc.type.driver.none.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
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https://purl.org/coar/version/c_ab4af688f83e57aa |
| format |
https://purl.org/coar/resource_type/c_7a1f |
| dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/20.500.12495/15703 |
| dc.identifier.instname.spa.fl_str_mv |
instname:Universidad El Bosque |
| dc.identifier.reponame.spa.fl_str_mv |
reponame:Repositorio Institucional Universidad El Bosque |
| dc.identifier.repourl.none.fl_str_mv |
repourl:https://repositorio.unbosque.edu.co |
| url |
https://hdl.handle.net/20.500.12495/15703 |
| identifier_str_mv |
instname:Universidad El Bosque reponame:Repositorio Institucional Universidad El Bosque repourl:https://repositorio.unbosque.edu.co |
| dc.language.iso.fl_str_mv |
spa |
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spa |
| dc.relation.references.none.fl_str_mv |
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Escobar Márquez, Lina MaríaBendahan Álvarez, Zita CarolinaCastaño Duque, Sandra PatriciaCalderón García, ElianaRamírez Robles, Luz Eliana2025-08-12T14:15:03Z2025-08-12T14:15:03Z2025-08https://hdl.handle.net/20.500.12495/15703instname:Universidad El Bosquereponame:Repositorio Institucional Universidad El Bosquerepourl:https://repositorio.unbosque.edu.coAntecedentes: Estudios acerca de ingeniería tisular establecen que los co-cultivos directos han sido útiles para comprender la interacción entre células debido a que la comunicación intercelular promueve el intercambio de señales y factores de crecimiento que favorecen la diferenciación hacia distintos linajes celulares. Adicionalmente, en otros estudios se reportan interacciones celulares en co-cultivos indirectos que proporcionan un panorama favorable para la generación de tejidos o la reconstrucción de defectos óseos. No obstante, no existe evidencia clara que compare los co-cultivos directos y los indirectos en el desarrollo de procesos de proliferación y diferenciación de células madre a OBs. Esto podría en un futuro establecer nuevas terapéuticas que brindan soluciones a la reparación de tejidos de diferentes patologías. Objetivo: Determinar y comparar el efecto en la diferenciación, proliferación y morfología de células madre mesenquimales (MSCs) derivadas de la pulpa dental (hDPSC) y pre-osteoblastos (Saos-2) co-cultivados de forma directa e indirecta in vitro. Materiales y Métodos: Se evaluó proliferación, viabilidad, morfología y diferenciación celular a partir de co- cultivo directo e indirecto de células de la pulpa dental (hDPSC) y Saos-2 en medio DMEM a los 7 , 14 y 21 días. Se utilizó control positivo con medio de diferenciación y control negativo con medio DMEM. Resultados: Se realizó inmunofenotipificación a las hDPSC donde se encontró células positivas para marcadores de superficie CD105, CD90, CD73 y se evaluaron características morfológicas donde se observó células fibroblastoides adheridas a la caja. El co-cultivo indirecto y el directo de células Saos-2+hDPSC, produjo un incremento significativo (P < 0.05) en el número de células y proliferación celular desde el 6to día en el co-cultivo indirecto y desde el cuarto día en el co-cultivo directo. En las células hDPSC y Saos-2 sin co-cultivo (control) no se observó tinción con el rojo de alizarina, mientras que las células tratadas con MDO como control positivo de diferenciación osteoblástica mostraron igualmente formación de nódulos de calcio. La mayor cantidad de rojo de alizarina como medida de absorbancia, se evidenció en las células co-cultivadas de forma indirecta y en las células directamente co-cultivadas . Estas absorbancias fueron significativamente mayores que las obtenidas en las células hDPSC y Saos-2 sin co-cultivo. Al analizar los cambios en la expresión de genes de diferenciación osteoblástica como RUNX2 y osteocalcina (OCN) se encontró que a los 7 días RUNX2 aumentó su expresión en todos los grupos experimentales excepto en las células hDPSC co-cultivadas de forma indirecta con Saos-2. Por otro lado, El mayor incremento en la expresión se evidenció en el co-cultivo directo (116 veces más que el control) y hDPSC tratadas con MDO (83 veces más que el control). La expresión se redujo significativamente en todos los grupos experimentales a los 21 días. Conclusiones: Este estudio sugiere que el co-cultivo con células Saos-2 y hDPSC tanto directo como indirecto es efectivo para inducir cambios morfológicos, en la proliferación y diferenciación osteoblástica. Por este motivo, pueden ser una herramienta útil en la investigación de terapias de regeneración para la reparación de defectos óseos.Grupo de investigación UMIMC - Unidad de manejo integral de malformaciones craneofacialesEspecialista en OrtodonciaEspecializaciónBackground: Studies on tissue engineering have demonstrated that direct co-cultures are useful for understanding cell interactions, as intercellular communication facilitates the exchange of signals and growth factors that promote differentiation into various cell lineages. Additionally, other studies report cellular interactions in indirect co-cultures that provide a favorable outlook for tissue generation or bone defect reconstruction. However, there is no clear evidence comparing direct and indirect co-cultures in the development of proliferation and differentiation processes of stem cells into OBs. This could, in the future, lead to the development of new therapies that provide solutions for tissue repair in various pathologies. Aim: To determine and compare the effect on the differentiation, proliferation, and morphology of mesenchymal stem cells (MSCs) derived from dental pulp (hDPSC) and pre-osteoblasts (Saos-2) co-cultured directly and indirectly in vitro. Methods: Cell proliferation, viability, morphology, and differentiation were evaluated from direct and indirect co-culture of dental pulp cells (hDPSC) and Saos-2 in DMEM medium at 7, 14, and 21 days. A positive control with differentiation medium and a negative control with DMEM medium were used. Results: Immunophenotyping was performed on hDPSC, where cells positive for surface markers CD105, CD90, and CD73 were identified, and morphological characteristics were evaluated, revealing that fibroblast-like cells adhered to the dish. The indirect and direct co-culture of Saos-2+hDPSC cells resulted in a significant increase (P < 0.05) in cell number and proliferation from day 6 in the indirect co-culture and from day 4 in the direct co-culture. In hDPSC and Saos-2 cells without co-culture (control), no alizarin red staining was observed, while cells treated with MDO as a positive control for osteoblastic differentiation also showed calcium nodule formation. The highest amount of alizarin red, as a measure of absorbance, was evident in both indirectly co-cultured cells and directly co-cultured cells. These absorbances were significantly higher than those obtained in hDPSC and Saos-2 cells without coculture. When analyzing changes in the expression of osteoblastic differentiation genes such as RUNX2 and osteocalcin (OCN), it was found that at 7 days, RUNX2 expression increased in all experimental groups except in hDPSC cells indirectly co-cultured with Saos-2. On the other hand, the greatest increase in expression was observed in direct co-culture (116 times higher than the control) and hDPSC treated with MDO (83 times higher than the control). Expression was significantly reduced in all experimental groups at 21 days. Conclusions: This study suggests that both direct and indirect co-culture with Saos-2 cells and hDPSC is effective in inducing morphological changes, proliferation, and osteoblastic differentiation. For this reason, they may be a useful tool in the investigation of regenerative therapies for bone defect repair.application/pdfAttribution-NonCommercial-ShareAlike 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-sa/4.0/Acceso abiertohttps://purl.org/coar/access_right/c_abf2http://purl.org/coar/access_right/c_abf2PreosteoblastosCélulas madrePulpa dentalOsteoblastosTécnicas de cultivoDiferenciación celularPre-osteoblastsStem cellsDental PulpOsteoblastsCulture techniquesCell differentiationWU400Evaluación de la diferenciación de células mesenquimales y pre-osteoblastos en un sistema de co-cultivo directo e indirectoEvaluation of the differentiation of mesenchymal cells and pre-osteoblasts in a direct and indirect co-culture systemEspecialización en OrtodonciaUniversidad El BosqueFacultad de OdontologíaTesis/Trabajo de grado - Monografía - Especializaciónhttps://purl.org/coar/resource_type/c_7a1fhttp://purl.org/coar/resource_type/c_7a1finfo:eu-repo/semantics/bachelorThesishttps://purl.org/coar/version/c_ab4af688f83e57aa1.Lee YC, Chan YH, Hsieh SC, Lew WZ, Feng S-W. 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