| Summary: | 碩士 === 國立中正大學 === 化學工程所 === 97 === This research manufactured three-dimensionally chitosan-gelatin scaffolds by the methods of inverted colloidal crystal and freeze-drying,then cultured with rat bone marrow stromal cells after 7-day. Firstly, the analysis of MTT assay indicated that BMSCs cultured in chitosan-gelatin
scaffolds had better cells proliferation than pure chitosan or pure gelatin scaffolds. The final cells population compared with initials can increase about 2.5 times. Secondly, Immunocytochemical staining of the BMSCs
cultured with basic medium in different mass ratio of freeze-drying or inverted colloidal crystal chitosan-gelatin scaffolds after 7-day cultivation with antibodies directed against cell surface marker CD44 and CD90 were detected by flow cytometer, then we showed that the percentage of BMSCs in ICC scaffolds was equally 62.88% but in freeze-drying scaffolds was equally 55.76%, and that the percentage of BMSCs will reduce when the higher content of chitosan blend into scaffolds. So, the results showed that ICC scaffolds can maintain the morphological of
BMSCs and keep the ability of self-renewal、potency. In addition, BMSCs cultured with basic medium in chitosan-gelatin scaffolds conjugated with laminin-derived peptides (123.5±13.25 μg/cm3) were recognized by flow cytometer then showed the percentage of BMSCs reduced to 27.60%. Furthermore, we also showed that laminin-derived peptides not only increase cells adhesive to 65%~75% but also slightly induced BMSCs differentiation into neuron-liked cells by immunofluorescent staining with antibodies directed against marker anti-NeuN. Finally, we used three different therapy ways to cue rat spinal cord injury after 2w，4w treatment. The results of Nissl staining showed
that implanted BMSCs into laminin-derived peptides conjugated with ICC scaffolds and then put it into the injury of spinal cord can achieve the better repair to neurons.
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