RESUMO
The aim of the present study was to explore the feasibility of the construction of engineered myocardial tissues in vitro with cardiomyocytelike cells derived from bone marrow mesenchymal stem cells (BMMSCs) and a polylacticcoglycolic acid (PLGA) polymer. The PLGA polymer was sheared into square pieces (10x10x1 mm), sterilized by Co60 irradiation, and hydrated in Dulbecco's modified Eagle's medium for 1 h. BMMSCs were isolated from the bone marrow of SpragueDawley rats and the third passage cells were induced by 5azacytidine (5aza). Following successful induction, the cells were trypsinized and suspended at a density of 1x109/ml. Then, the cell suspension was added to the PLGA scaffold and cultured for 14 days. The morphological changes of BMMSCs were observed using phase contrast microscopy. Immunofluorescence staining was used to identify the cardiomyocytelike cells. Hematoxylin and eosin (H&E) and immunohistochemical staining were used to observe the morphology of the engineered myocardial tissues. The cell adhesion rates and scanning electron microscopy were used to observe the compatibility of the cardiomyocytelike cells and PLGA. Transmission electron microscopy was used to view the ultrastructure of the engineered myocardial tissues. BMMSCs in primary culture presented round or short spindle cell morphologies. Following induction by 5aza, the cells exhibited a long spindle shape and a parallel arrangement. Analysis of the cell adhesion rates demonstrated that the majority of the cardiomyocytelike cells had adhered to the PLGA scaffolds at 24 h. H&E staining suggested that the cardiomyocytelike cells with spindle nuclei were evenly distributed in the PLGA scaffold. Immunofluorescence staining revealed that the cardiomyocytelike cells were positive for cardiac troponin I. Scanning electron microscopy demonstrated that the inoculated cells were well attached to the PLGA scaffold. Transmission electron microscopy indicated that the engineered myocardial tissues contained wellarranged myofilaments, desmosomes, gap junction and Z linelike structures. The present study successfully constructed engineered myocardial tissues in vitro with a PLGA polymer and cardiomyocytelike cells derived from BMMSCs, which are likely to share various structural similarities with the original heart tissue.
Assuntos
Células-Tronco Mesenquimais/citologia , Miócitos Cardíacos/citologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Diferenciação Celular , Células Cultivadas , Masculino , Miocárdio/citologia , Ratos Sprague-DawleyRESUMO
This study investigated the potential application of a zirconium oxide (ZrO2) ceramic foam culturing system to the production of glial cell line-derived neurotrophic factor (GDNF). Three sets of ZrO2 ceramic foams with different pore densities of 10, 20, and 30 pores per linear inch (PPI) were prepared to support a 3D culturing system. After primary astrocytes were cultured in these systems, production yields of GDNF were evaluated. The biomaterial biocompatibility, cell proliferation and activation of cellular signaling pathways in GDNF synthesis and secretion in the culturing systems were also assessed and compared with a conventional culturing system. In this study, we found that the ZrO2 ceramic foam culturing system was biocompatible, using which the GDNF yields were elevated and sustained by stimulated cell proliferation and activation of signaling pathways in astrocytes cultured in the system. In conclusion, the ZrO2 ceramic foam is promising for the development of a GDNF mass production device for Parkinson's disease treatment.