RESUMEN
Bone marrow stromal cells are multipotential stem cells that contribute to the differentiation of tissues such as bone, cartilage, fat and muscle. In the experiment, we found that bone marrow stromal cells can be induced to differentiate into cells expressing characteristic markers of Schwann cells, such as S-100 and glial fibrillary acidic protein, promoting peripheral nerve regeneration. Tissue-engineered bioartificial nerve grafting of rats by differentiated bone marrow stromal cells was applied for bridging a 10 mm-long sciatic nerve defect. Twenty-eight inbred strains of female F344 rats weighing 160 approximately 200 g were randomly divided into four nerve grafting groups, with seven rats in each group. Differentiated bone marrow stromal cell-laden group: poly(lactic-co-glycolic) acid tubes with an intrinsic framework were seeded with syngeneic bone marrow stromal cells which were induced for 5 days; Schwann cell-laden group: poly(lactic-co-glycolic) acid tubes with an intrinsic framework were seeded with syngeneic Schwann cells; acellular group: poly(lactic-co-glycolic) acid tubes were only filled with an intrinsic framework; autografts group. Three months later, a series of examinations was performed, including electrophysiological methods, walking track analysis, immunohistological staining of nerves, immunostaining of S-100 and neurofilament, and axon counts. The outcome indicated that bone marrow stromal cells are able to differentiate into Schwann-like cells and Schwann-like cells could promote nerve regeneration. Bone marrow stromal cells may be potentially optional seed cells for peripheral nerve tissue engineering because of abilities of promoting axonal regeneration.
Asunto(s)
Células de la Médula Ósea/fisiología , Trasplante de Médula Ósea , Diferenciación Celular/fisiología , Células de Schwann/fisiología , Ingeniería de Tejidos/métodos , Animales , Animales Recién Nacidos , Western Blotting/métodos , Células Cultivadas , Estimulación Eléctrica/métodos , Proteína Ácida Fibrilar de la Glía/metabolismo , Inmunohistoquímica/métodos , Microscopía Electrónica de Transmisión/métodos , Conducción Nerviosa/fisiología , Conducción Nerviosa/efectos de la radiación , Proteínas de Neurofilamentos/metabolismo , Ratas , Ratas Endogámicas F344 , Proteínas S100/metabolismo , Células de Schwann/ultraestructura , Factores de TiempoRESUMEN
OBJECTIVE: To investigate the effect of transforming growth factor (TGF) beta 1 gene transfection on the growth of mesenchymal stem cells(MSCs) and to evaluate a new biomimetic biodegradable polymer as scaffolds for applications in articular cartilage tissue engineering. METHODS: Principles of tissue engineering were combined organically with principles of gene therapy to produce cultured periosteum-derived MSCs transduced with the full-length rat TGF-beta 1 cDNA in vitro. These cells were then seeded onto three-dimensional porous poly-DL-lactide scaffolds modified with poly-L-lysine that mimicked cell-binding domains found on natural extracellular matrix to promote specific cell adhesion. The adhesion, proliferation, and differentiation of the transfected MSCs were examined with scanning electron microscope within 2 weeks. RESULTS: All cells adhered to the biomimetic matrices well, but more cartilage-like tissue was formed for TGF-beta 1 gene modified MSCs/scaffolds composites than for the control groups. Transfer of gene encoding TGF-beta 1 to MSCs promoted its proliferation and differentiation significantly. CONCLUSIONS: The TGF-beta 1 gene transduced MSCs/biomimetic matrix composites used in this study was the first attempt to apply the principles of molecular tissue engineering for articular cartilage repair. This new molecular tissue engineering approach could be of potential benefit to repair damaged articular cartilage, especially in osteoarthritis. The new biomimetic biodegradable polymer matrices modified with biomolecules not only have good structural compatibility, but also have better interfacial compatibility and bioactivity, and can be used as scaffolds for articular cartilage tissue engineering.
Asunto(s)
Materiales Biocompatibles , Polímeros , Células Madre/citología , Ingeniería de Tejidos/métodos , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/farmacología , Animales , Adhesión Celular , Técnicas de Cultivo de Célula/métodos , Diferenciación Celular , División Celular , Ácido Láctico , Mesodermo/citología , Conejos , TransfecciónRESUMEN
In view of the development of miniaturized sensor arrays, a solid-contact ammonium ion selective electrode has been investigated. A conductive polypyrrole film was electrochemically deposited on a glassy carbon surface and used as an internal solid contact layer between the sensing membrane and solid electrode surface. A systematic evaluation of the important parameters affecting the electromotive force (emf) response is presented. The performances of this solid-contact sensor were verified using a batch-mode measurement setup and a wall-jet flow cell system. The designed sensor exhibited excellent selectivity for the primary ion and a linear response over the pNH4+ range 1-5 with a slope of 56.3 mV decade(-1) . The sensor has a fast response and is relatively robustness, and was also used to determine ammonium concentrations in natural waters, with promising results.