| Summary: | 碩士 === 國立成功大學 === 化學工程學系碩博士班 === 100 === Skeletal muscle tissue is compared of bundles of highly oriented and densely packed muscle fibers, each with multinucleated myotubes derived from myoblasts. Therefore, the process of myotube formation from myoblasts plays an important role for skeletal muscle tissue engineering. The goal of this study was to examine the differentiation of skeletal myoblasts on aligned and electrically conductive nanofibrous scaffolds, and then investigate the effect of electrical pulse stimulation (EPS) combined with this scaffolds on myotubes maturation and functional sarcomere assembly. The aligned and electrically conductive nanofibers composed of poly-ε-caprolactone (PCL) and polyaniline (PANi) were fabricated by a magnetic-field-assisted electrospinning (MFAES) method. The diameter of Aligned-PCL/PANi nanofibers was 307.6 ± 39.6 nm with highly oriented structure (〉 90% fibers within ± 10° of the preferred direction). The cell viability results indicated that Aligned-PCL/PANi nanofibers induced myoblasts alignment and significantly enhanced cell proliferation compared to Aligned-PCL nanofibers. After differentiation for 5 days, the myotube number and length, fusion index and degree of maturation of Aligned-PCL/PANi were greater (38.9 ± 2.0, 198.3 ± 22.2 μm, 36.7 ± 1.3% and 53.2 ± 2.2%) than that of Random-PCL/PANi (30.3 ± 2.4, 147.2 ± 30.7 μm, 27.9 ± 3.2% and 44.2 ± 3.0%). Finally, we investigated the effect of EPS on myotube maturation and sarcomere assembly on the Aligned-PCL/PANi. We found EPS could enhance the myotube formation and maturation. The myotube number and length, fusion index and maturation index of myocytes on Aligned-PCL/PANi with EPS at day 9 for 12 h were greater (76.3 ± 2.1, 425.5 ± 54.2 μm and 86.0 ± 2.2%) than that without EPS (65.7 ± 3.3, 369.4 ± 30.2 μm and 80.3 ± 1.6%). Additionally, when EPS was applied at day 7 for 24 h, the percentage of striated myotubes can reach to 47%. These results indicate that aligned and electrically conductive nanofibers can modulate the induction of myoblasts into mature and striated myotubes when EPS was applied, suggesting that the combined strategy have great potential for skeletal muscle tissue engineering.
|
|---|
