Mulder M M, Hitchcock R W, Tresco P A
University of Utah, Department of Bioengineering, Salt Lake City 84112, USA.
J Biomater Sci Polym Ed. 1998;9(7):731-48. doi: 10.1163/156856298x00118.
Studies geared towards understanding the interaction between skeletal muscle and biomaterials may provide useful information for the development of various emerging technologies, ranging from novel delivery vehicles for genetically modified cells to fully functional skeletal muscle tissue. To determine the utility of elastomeric materials as substrates for such applications, we asked whether skeletal myogenesis would be supported on a commercially available polyurethane, Tecoflex SG-80A. G8 skeletal myoblasts were cultured on Tecoflex two-dimensional solid thin films fabricated by a spin-casting method. Myoblasts attached, proliferated, displayed migratory activity and differentiated into multinucleated myotubes which expressed myosin heavy chain on solid thin films indicating that Tecoflex SG-80A was permissive for skeletal myogenesis. Porous three-dimensional (3-D) cell scaffolds were fabricated in a variety of shapes, thicknesses, and porosities by an immersion precipitation method, and where subsequently characterized with microscopic and mechanical methods. Mechanical analysis revealed that the constructs were elastomeric, recovering their original length following 100% elongation. The 3-D substrates were seeded with muscle precursors to determine if muscle differentiation could be obtained within the porous network of the fabricated constructs. Following several weeks in culture, histological studies revealed the presence of multinucleated myotubes within the elastomeric material. In addition, immunohistochemical analysis indicated that the myotubes expressed the myosin heavy chain protein suggesting that the myotubes had reached a state of terminal differentiation. Together the results of the study suggest that it is indeed feasible to engineer bioartificial systems consisting of skeletal muscle cultivated on a 3-D elastomeric substrate.
旨在理解骨骼肌与生物材料之间相互作用的研究，可能会为各种新兴技术的发展提供有用信息，这些技术涵盖从用于基因改造细胞的新型递送载体到全功能骨骼肌组织等多个领域。为了确定弹性体材料作为此类应用底物的效用，我们探究了在市售聚氨酯Tecoflex SG - 80A上是否能支持骨骼肌生成。将G8骨骼肌成肌细胞培养在通过旋铸法制备的Tecoflex二维固体薄膜上。成肌细胞附着、增殖、表现出迁移活性并分化为多核肌管，这些肌管在固体薄膜上表达肌球蛋白重链，表明Tecoflex SG - 80A有利于骨骼肌生成。通过浸没沉淀法制备了各种形状、厚度和孔隙率的多孔三维（3 - D）细胞支架，随后用显微镜和力学方法对其进行表征。力学分析表明，这些构建体具有弹性，在100%伸长后能恢复其原始长度。将肌肉前体细胞接种到3 - D底物上，以确定在制备的构建体的多孔网络内是否能实现肌肉分化。培养数周后，组织学研究显示弹性体内存在多核肌管。此外，免疫组织化学分析表明肌管表达肌球蛋白重链蛋白，这表明肌管已达到终末分化状态。该研究结果共同表明，构建由三维弹性体底物上培养的骨骼肌组成的生物人工系统确实是可行的。
