Evaluación in vitro de la adhesión de células troncales mesenquimales a matrices dentales impresas en tercera dimensión / In Vitro Evaluation of the Adhesion of Mesenchymal Stem Cells to Three-Dimensional Printed Matrices

Álvaro Andrés Rodríguez Sáenz, María Alejandra Lozano Macías, Astrid Eugenia Benedetti Canabal, Juan Carlos Munévar Niño, Jorge Alberto Sarmiento O'Meara, Roberto Alfonso Gómez Bahamón

Abstract


RESUMEN. Antecedentes: En la ingeniería de tejidos es fundamental estudiar el sinergismo entre las células troncales mesenquimales y el biomaterial para tener un mayor control sobre los biomiméticos. De esto depende el éxito de tratamientos de lesiones de tamaño crítico. Objetivo: Evaluar la adhesión celular in vitro de células troncales de la pulpa dental humana (hDPSCs) en matrices impresas con ácido poliláctico (APL). Métodos: Se utilizaron muestras de hDPSCs criopreservadas y expandidas, cultivadas sobre 24 matrices dentales impresas 3D en APL durante 1, 7 y 15 días. Se evaluó la fenotipificacion de la hDPSCs por citometría de flujo y la adhesión celular a la matriz por medio de microscopio electrónico de barrido (SEM). Los datos fueron reportados en porcentajes, tanto para marcador analizado, como para la cantidad de células adheridas. Resultados: hDPSCs expresaron positivamente anticuerpos CD73 y CD90 de casi 100 % y CD105 de 56,7 %. Así mismo, expresaron negativamente, anticuerpos CD34 y CD45 mayor a 98 %. Se observó en SEM que a los 15 días el 99,88 % de las hDPSCs presentaron forma fusiforme o estrellada lo que significa que estas células se adhirieron a la matriz de APL. Conclusión: El APL no es citotóxico para las hDPSCs por su composición y características biocompatibles, lo que proporcionó que las células se adhirieron y proliferaron sobre la matriz dental impresa en 3D demostrando ser un método in vitro efectivo para emplear en futuros estudios de regeneración de tejidos en odontología.

ABSTRACT. Background: The study of synergy between mesenchymal stem cells and biomaterial in tissue engineering is fundamental in order to have greater control over the biomimetics for the success of critical clinical treatments depends on this. Objective: to evaluate in vitro cellular adhesion of human dental pulp stem cells (hDPSCs) in three-dimensional printed matrices synthesized with poly-lactic acid (PLA). Method: The study used passage-expanded cryopreserved dental stem cell samples cultivated on 24 three-dimensional dental matrices of PLA during 1, 7 and 15 days. Phenotypification of the DMSC was carried out with flow cytometry and the cellular adhesion to the matrix with morphological analysis using an SEM electron microscope. Data were reported in percentages for each marker in the phenotypification as well as for the amount of adhered cells per time period. Results: The hDPSCs expressed positively, CD73 and CD 90 antibodies of almost 100 % while the CD 105 only had 56, 7 %. The cells presented negative expression of the CD34 and CD45 antibodies in more than 98 %. The SEM showed that 99, 88 % of the DMSC had a fusiform or star shape after 15 days meaning that they had adhered to the matrix. Conclusion: PLA biomaterial is not cytotoxic for the DMSC due to its composition and biocompatible characteristics, which helped the cells, adhere themselves and proliferate on the matrix. This proved to be an effective in vitro method, which may be used in future studies of critical- size tissue lesion studies in dentistry.

 


Keywords


diseño industrial; ingeniería tisular; odontología; ácido poliláctico; biomateriales; células madre; impresión 3D; pulpa dental; regeneración tisular

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DOI: http://dx.doi.org/10.11144/Javeriana.uo36-77.eact

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