Secreción de factores de crecimiento y citoquinas, durante la producción de tejido artificial bajo dos concentraciones diferentes de oxígeno en un biorreactor tipo Spinner
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Convencionalmente, pérdidas grandes de tejido, aquellas que no se pueden suturar, se tratan con autoinjertos, provenientes del propio paciente, aloinjerto, tejidos compatibles de la misma especie, o xenoinjertos, provenientes de otras especies. Sin embargo, tienen problemas respecto a la disponibilidad, rechazo y transmisión de enfermedades. Por ello, la ingeniería de tejidos y la medicina regenerativa desarrollan terapias que reemplacen, restablezcan o mejoren las funciones de tejidos lesionados o perdidos, conjugando conocimientos de ingeniería y ciencias biomédicas. El uso de medios condicionados, secretados por células relacionadas con la reparación de la herida, surge como alternativa viable desde el punto de vista de manufactura, costos, manipulación y distribución. En este trabajo se cultivaron fibroblastos sobre soportes de colágeno I en un biorreactor tipo spinner, bajo dos condiciones diferentes de oxígeno (hipoxia y normoxia), proporcionando a las células el estímulo ambiental y soporte necesario para producir tejido artificial y secreción de factores de crecimiento y citoquinas relacionadas en el proceso de reparación de heridas. Estos factores fueron medidos junto con la producción de lactato y consumo de glucosa durante el cultivo, después de la formación del tejido artificial. Al revisar ambos medios se encontró que el medio producido bajo ambas condiciones presentaba secreción de proteínas proangiogénicas, provascularizantes, anti y pro inflamarorias, lo cual sugiere que poseen un perfil adecuado al esperado para cierre de heridas.
Abstract. Conventionally, when large portions of tissue are lost, those that cannot be sutured, are treated with autografts from patient, allografts, compatible tissues from the same specie, or xenografts, from other species. However, those have problems with availability, rejection and diseases transmission. Therefore, tissue engineering and regenerative medicine develop therapies to replace, reestablish or improve functions of lost or damage tissues, combining engineering and biomedical sciences knowledge. Usage of conditioned media, secreted by cells related to wound healing, emerges as a viable alternative from the manufacturing, cost, manipulation and distribution point of view. In this work fibroblasts were cultured on collagen I scaffolds in a spinner bioreactor, under two different oxygen conditions (hypoxia and normoxia), providing the support and necessary environmental stimulation to cells, to produce artificial tissue and growth factor and cytokines secretion related to wound healing. These factors were measured along with lactate production and glucose consumption during culture, after artificial tissue formation. After analyzing the media, it was found that media produced under both conditions showed secretion of proangiogenic, provascular and anti and proinflammatory proteins, suggesting the adequate profile for wound healing.
Abstract. Conventionally, when large portions of tissue are lost, those that cannot be sutured, are treated with autografts from patient, allografts, compatible tissues from the same specie, or xenografts, from other species. However, those have problems with availability, rejection and diseases transmission. Therefore, tissue engineering and regenerative medicine develop therapies to replace, reestablish or improve functions of lost or damage tissues, combining engineering and biomedical sciences knowledge. Usage of conditioned media, secreted by cells related to wound healing, emerges as a viable alternative from the manufacturing, cost, manipulation and distribution point of view. In this work fibroblasts were cultured on collagen I scaffolds in a spinner bioreactor, under two different oxygen conditions (hypoxia and normoxia), providing the support and necessary environmental stimulation to cells, to produce artificial tissue and growth factor and cytokines secretion related to wound healing. These factors were measured along with lactate production and glucose consumption during culture, after artificial tissue formation. After analyzing the media, it was found that media produced under both conditions showed secretion of proangiogenic, provascular and anti and proinflammatory proteins, suggesting the adequate profile for wound healing.