| 研究実績の概要 |
Methylcellulose (MC) based scaffolds were prepared with MC (500mg) mixed with gelatin (50 mg) and agarose (50 mg) in 25 mL dimethyl sulfoxide (DMSO) and 5g ground-NaCl by vigorous stirring for 4 h. Cross-linking agent (carbonyl diimidazole, CDI) was added and vigorously agitated for 15 min. Solutions were incubated in ice bath for scaffold formation, and then washed with distilled water 5 times to remove NaCl, DMSO and unreacted reagents. Finally, these samples were frozen and lyophilized. Well dishes of 35mm, 24mm and 96 well, were used as molds to obtain a variety sized with a consistent disk shape.
During lyophilization, some of the samples presented cracks that compromised the integrity of the scaffold during its removal from the molds. The use of a tissue layer to fix the gel within the wells partially solved this problem.
Results also showed that NaCL crystal greatly influenced the scaffold porosity. With less concentration, reduced porosity was observed. Moreover, pore size was rather inconsistent throughout the experiment. We also found that concentration of NaCl influenced the freezing point, therefore Salt, DMSO and unreacted chemicals had to be carefully removed with distilled water before freezing at -80C to avoid damages to the gel matrixes. Moreover, salinity induces changes in cell membrane stability and could affect cell proliferation. Although MC scaffolds were achieved, thickness, pore size and salinity were rather inconsistent. This has strong influence cell culture results, therefore more testing is required.
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