Examinando por Autor "Carrillo-Delgado, Esmeralda"
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Ítem A soft 3D polyacrylate hydrogel recapitulates the cartilage niche and allows growth-factor free tissue engineering of human articular cartilage(Elsevier, 2019) Jiménez-González, Gema; Venkateswaran, Seshasailam; López-Ruiz, Elena; Perán, Macarena; Pernagallo, Salvatore; Díaz-Manchón, Juan José; Canadas, Raphaël; Antich, Cristina; Oliveira, Joaquim Miguel; Callanan, Anthony; Wallace, Robert; Reis, Rui L.; Montañez, Elvira; Carrillo-Delgado, Esmeralda; Bradley, Mark; Marchal, Juan AntonioCartilage degeneration or damage treatment is still a challenge, but, tissue engineering strategies, which combine cell therapy strategies, which combine cell therapy and scaffolds, and have emerged as a promising new approach. In this regard, polyurethanes and polyacrylates polymers have been shown to have clinical potential to treat osteochondral injuries. Here, we have used polymer microarrays technology to screen 380 different polyurethanes and polyacrylates polymers. The top polymers with potential to maintain chondrocyte viability were selected, with scale-up studies performed to evaluate their ability to support chondrocyte proliferation during long-term culture, while maintaining their characteristic phenotype. Among the selected polymers, poly (methylmethacrylate-co methacrylic acid), showed the highest level of chondrogenic potential and was used to create a 3D hydrogel. Ultrastructural morphology, microstructure and mechanical testing of this novel hydrogel revealed robust characteristics to support chondrocyte growth. Furthermore, in vitro and in vivo biological assays demonstrated that chondrocytes cultured on the hydrogel had the capacity to produce extracellular matrix similar to hyaline cartilage, as shown by increased expression of collagen type II, aggrecan and Sox9, and the reduced expression of the fibrotic marker’s collagen type I. In conclusion, hydrogels generated from poly (methylmethacrylate-co-methacrylic acid) created the appropriate niche for chondrocyte growth and phenotype maintenance and might be an optimal candidate for cartilage tissue-engineering applications.Ítem Activin A/BMP2 chimera AB235 drives efficient redifferentiation of long term cultured autologous chondrocytes(Nature Research, 2015) Jiménez-González, Gema; López-Ruiz, Elena; Kwiatkowski, Witek; Montañez, Elvira; Arrebola, Francisco; Carrillo-Delgado, Esmeralda; Gray, Peter; Izpisua, Juan Carlos; Choe, Senyon; Perán, Macarena; Marchal, Juan AntonioAutologous chondrocyte implantation (ACI) depends on the quality and quantity of implanted cells and is hindered by the fact that chondrocytes cultured for long periods of time undergo dedifferentiation. Here we have developed a reproducible and efficient chondrogenic protocol to redifferentiate chondrocytes isolated from osteoarthritis (OA) patients. We used morphological, histological and immunological analysis together with a RT-PCR detection of collagen I and collagen II gene expression to show that chondrocytes isolated from articular cartilage biopsies of patients and subjected to long-term culture undergo dedifferentiation and that these cells can be redifferentiated following treatment with the chimeric Activin A/BMP2 ligand AB235. Examination of AB235-treated cell pellets in both in vitro and in vivo experiments revealed that redifferentiated chondrocytes synthesized a cartilage-specific extracellular matrix (ECM), primarily consisting of vertically-orientated collagen fibres and cartilage-specific proteoglycans. AB235 treated cell pellets also integrated into the surrounding subcutaneous tissue following transplantation in mice as demonstrated by their dramatic increase in size while non-treated control pellets disintegrated upon transplantation. Thus, our findings describe an effective protocol for the promotion of redifferentiation of autologous chondrocytes obtained from OA patients and the formation of a cartilage-like ECM that can integrate into the surrounding tissue in vivo.Ítem Cellular extracts from post-mortem human cardiac tissue direct cardiomyogenic differentiation of human adipose tissue-derived stem cells(Elsevier, 2010) Perán, Macarena; Marchal, Juan Antonio; López-Ruiz, Elena; Jiménez Navarro, Manuel; Boulaiz, Hourai; Rodríguez-Serrano, Fernado; Carrillo-Delgado, Esmeralda; Sánchez-Espín, Gema; de-Teresa, Eduardo; Tosh, David; Aránega, AntoniaBackground aims. The goal was to induce the transdifferentiation (or conversion) of human adipose-derived stem cells to cardiomyocytes using an intracellular extract obtained from adult human heart tissue. Methods. Human adult stem cells from lipoaspirates were transiently permeabilized, exposed to human atrial extracts and allowed to recover in culture. Results. After 21 days, the cells acquired a cardiomyocyte phenotype, as demonstrated by morphologic changes (appearance of binucleate, striated cells and branching fibers), immunofl uorescence detection of cardiac-specifi c markers (connexin-43, sarcomeric a-actinin, cardiac troponin I and T, and desmin) and the presence of cardiomyocyte-related genes analyzed by reverse transcription – polymerase chain reaction (cardiac myosin light chain 1, a -cardiac actin, cardiac troponin T and cardiac b -myosin). Conclusions. We have demonstrated for the fi rst time that adult cardiomyocytes obtained from human donors retain the capacity to induce cardiomyocyte differentiation of mesenchymal stromal cells. The use of autologous extracts for reprogramming adult stem cells may have potential therapeutic implications for treating heart disease.Ítem Human cardiac tissue induces transdifferentiation of adult stem cells towards cardiomyocytes(Elsevier, 2010) Perán, Macarena; Marchal, Juan Antonio; López-Ruiz, Elena; Jiménez-Navarro, Manuel; Boulaiz, Houria; Rodríguez-Serrano, Fernando; Carrillo-Delgado, Esmeralda; Sánchez-Espín, Gema; de-Teresa, Eduardo; Tosh, David; Aránega, AntoniaBackground aims. The goal was to induce the transdifferentiation (or conversion) of human adipose-derived stem cells to cardiomyocytes using an intracellular extract obtained from adult human heart tissue. Methods. Human adult stem cells from lipoaspirates were transiently permeabilized, exposed to human atrial extracts and allowed to recover in culture. Results. After 21 days, the cells acquired a cardiomyocyte phenotype, as demonstrated by morphologic changes (appearance of binucleate, striated cells and branching fi bers), immunofl uorescence detection of cardiac-specifi c markers (connexin-43, sarcomeric a-actinin, cardiac troponin I and T, and desmin) and the presence of cardiomyocyte-related genes analyzed by reverse transcription – polymerase chain reaction (cardiac myosin light chain 1, a cardiac actin, cardiac troponin T and cardiac b-myosin). Conclusions. We have demonstrated for the fi rst time that adult cardiomyocytes obtained from human donors retain the capacity to induce cardiomyocyte differentiation of mesenchymal stromal cells. The use of autologous extracts for reprogramming adult stem cells may have potential therapeutic implications for treating heart disease.