Development of micro-patterned device incorporated with neurotrophic gradient and supportive Schwann cells for the applications in neural tissue engineering
| Autor: | Yeh Chia Wei, 葉佳偉 |
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| Rok vydání: | 2015 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 104 The current gold standard treatment in clinic for peripheral nerve injury is autologous nerve graft. Although there is no immune response after graft and complete extracellular matrix and Schwann cells are preserved in the nerve tissue, limited amount of available nerve and possible neuroma formation accompanied with permanent functional loss at donor site are the main concerns. In these years, the nerve guidance conduit has been developed as an alternative way to repair peripheral nerve. However, without incorporating proper stimulating factors, the prognosis is still not very satisfied. In this study, a biodegradable polymer (poly (glycerol-co-sebacic acid), PGS) based novel device has been developed and characterized. By incorporating three stimulating factors: 1) micro-patterned surface, that can directionally guide the axon as physical cue; 2) neurotrophic gradient membrane, that can continually attract axon outgrowth from the proximal to distal stump as chemical cue; 3) Schwann cells, that can support the growth of neurite and form myelin sheath around axon as biological cue, we expect that this scaffold can be used as a promising nerve guidance conduit for peripheral nerve regeneration. The results showed that the micro-patterned surface with specific dimensions of channels and chambers can be fabricated with good uniformity. Attachment and extension of differentiated neuron cells were observed in larger chamber area. The directional extension of neurite along direction of channel were also observed. By immunocytochemistry and flow cytometry, strong expression of s-100β and p75 low affinity receptor of nerve growth factor can be observed, indicating that Schwann cells can be successfully obtained from rat sciatic nerve and maintained their phenotype under in vitro culture environment. When co-culturing Schwann cells with neural stem cells, neural stem cells will differentiate toward neuronal cells. The gradient distribution of nerve growth factor 7S on gelatin membrane was successfully achieved. Significant enhancement of neurite area/field and length/field were also observed in higher concentration of NGF gelatin segment. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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