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Silkworm Silk Fiber Bundles as Improved In Vitro Scaffolds for Skeletal Muscle
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-11-19 , DOI: 10.1021/acsbiomaterials.0c00987 Matthew H Clegg 1 , Thomas I Harris 2 , Xiaoli Zhang 2 , Jacob T Barney 1 , Justin A Jones 2 , Elizabeth Vargis 1
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-11-19 , DOI: 10.1021/acsbiomaterials.0c00987 Matthew H Clegg 1 , Thomas I Harris 2 , Xiaoli Zhang 2 , Jacob T Barney 1 , Justin A Jones 2 , Elizabeth Vargis 1
Affiliation
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To mimic skeletal muscle tissues in vitro, native and transgenic spider silk/silkworm silks were seeded with C2C12 myoblasts to observe if these three-dimensional substrates are preferable to a traditional two-dimensional polystyrene cell culture surface. Silks were wound around an acrylic chassis to produce a novel, three-dimensional cell culture device with suspended muscle fibers that genetically and morphologically resemble native skeletal muscle tissue. The transgenic spider silk/silkworm silk has never before been studied for this application. Genetic expression verified skeletal muscle lineage and differentiation, while fluorescent imaging verified contractile protein synthesis. Genetic analysis also revealed an increase in expression of the Myh2 contractile protein gene on silkworm silks, particularly on the transgenic silk. Mechanical properties and protein secondary structure content of the silks indicated correlation between substrate properties and Myh2 gene expression. This increase in contractile protein gene expression suggests that biologically derived silk substrates that are suspended may be a preferable substrate for in vitro muscle modeling because of the proteinaceous character and mechanical flexibility of the silk.
中文翻译：
蚕丝纤维束作为骨骼肌体外支架的改进
为了在体外模拟骨骼肌组织，用 C2C12 成肌细胞播种天然和转基因蜘蛛丝/蚕丝，以观察这些三维底物是否优于传统的二维聚苯乙烯细胞培养表面。将丝绸缠绕在丙烯酸底盘上，以生产一种新颖的 3D 细胞培养装置，该装置具有悬浮的肌肉纤维，在遗传和形态上类似于天然骨骼肌组织。以前从未研究过转基因蜘蛛丝/蚕丝用于此应用。基因表达验证了骨骼肌谱系和分化，而荧光成像验证了收缩蛋白的合成。遗传分析还显示Myh2的表达增加蚕丝，特别是转基因蚕丝上的收缩蛋白基因。丝绸的机械性能和蛋白质二级结构含量表明底物特性与Myh2基因表达之间存在相关性。这种收缩蛋白基因表达的增加表明，由于蚕丝的蛋白质特征和机械柔韧性，悬浮的生物衍生丝底物可能是体外肌肉建模的优选底物。
更新日期：2020-12-14
中文翻译：
蚕丝纤维束作为骨骼肌体外支架的改进
为了在体外模拟骨骼肌组织，用 C2C12 成肌细胞播种天然和转基因蜘蛛丝/蚕丝，以观察这些三维底物是否优于传统的二维聚苯乙烯细胞培养表面。将丝绸缠绕在丙烯酸底盘上，以生产一种新颖的 3D 细胞培养装置，该装置具有悬浮的肌肉纤维，在遗传和形态上类似于天然骨骼肌组织。以前从未研究过转基因蜘蛛丝/蚕丝用于此应用。基因表达验证了骨骼肌谱系和分化，而荧光成像验证了收缩蛋白的合成。遗传分析还显示Myh2的表达增加蚕丝，特别是转基因蚕丝上的收缩蛋白基因。丝绸的机械性能和蛋白质二级结构含量表明底物特性与Myh2基因表达之间存在相关性。这种收缩蛋白基因表达的增加表明，由于蚕丝的蛋白质特征和机械柔韧性，悬浮的生物衍生丝底物可能是体外肌肉建模的优选底物。
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