Mehrotra Shreya, Nandi Samit Kumar, Mandal Biman B
Biomaterial and Tissue Engineering Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati-781039, Assam, India.
J Mater Chem B. 2017 Aug 21;5(31):6325-6338. doi: 10.1039/c7tb01494e. Epub 2017 Jul 28.
Heart failure, due to necrosis of heart tissue, interminably poses a significant burden on world-wide health care systems. In this context, a facile tissue engineering approach using mulberry (Bombyx mori) and non-mulberry (Antheraea assama) silk fibroin (SF) has been delved into. Amalgamating the efficacious attributes of cell sheet tissue engineering and robusticity of silk biomaterial, we developed a 3-D construct using silk films to promote cardiac tissue regeneration. Herein, the fabricated patterned silk films were physico-chemically characterized and analysed for their compatibility with cardiomyocytes. The presence of nanogrooves on silk films provided contact guidance to the growing cardiomyocytes allowing them to form unidirectionally aligned cell monolayers. Non-mulberry silk films exhibiting significantly greater mechanical strength and low immunogenicity in vitro and in vivo supported better growth, proliferation and maturation of both primary rat cardiomyocytes (PCMs) and H9c2 cells. The directional cue and presence of cell binding motifs such as RGD in A. assama films favoured the growth and maturation of cardiomyocytes to their functional phenotype. Cardiomyocyte maturation was attested by significant (p≤ 0.05) upregulation of myosin heavy chain-α (∼1.25 fold), connexin 43 (∼2 fold), and troponin I (∼1.25 fold) genes in PCMs grown on non-mulberry silk films. The patterned silk-cardiomyocyte monolayers were then stacked onto each other while maintaining their alignment to form a 3-D construct which exhibited structural integrity and uniform cellular distribution. Taking together, this work attests the suitability of non-mulberry A. assama silk fibroin as a potential biomaterial and prospects of exploring silk-cardiomyocyte monolayers for cardiac tissue engineering applications.
由于心脏组织坏死导致的心力衰竭，持续给全球医疗保健系统带来重大负担。在此背景下，人们深入研究了一种使用桑蚕丝（家蚕）和非桑蚕丝（柞蚕）丝素蛋白（SF）的简便组织工程方法。结合细胞片层组织工程的有效特性和丝生物材料的坚固性，我们使用丝膜开发了一种三维构建体，以促进心脏组织再生。在此，对制备的图案化丝膜进行了物理化学表征，并分析了它们与心肌细胞的相容性。丝膜上纳米凹槽的存在为生长中的心肌细胞提供了接触导向，使它们能够形成单向排列的细胞单层。非桑蚕丝膜在体外和体内均表现出显著更高的机械强度和低免疫原性，支持原代大鼠心肌细胞（PCM）和H9c2细胞更好地生长、增殖和成熟。柞蚕丝膜中RGD等细胞结合基序的定向线索和存在有利于心肌细胞生长并成熟为其功能表型。在非桑蚕丝膜上生长的PCM中，肌球蛋白重链-α（约1.25倍）、连接蛋白43（约2倍）和肌钙蛋白I（约1.25倍）基因显著上调（p≤0.05），证明了心肌细胞的成熟。然后将图案化的丝-心肌细胞单层相互堆叠，同时保持它们的排列，形成一个三维构建体，该构建体表现出结构完整性和均匀的细胞分布。综上所述，这项工作证明了非桑柞蚕丝素蛋白作为一种潜在生物材料的适用性，以及探索丝-心肌细胞单层用于心脏组织工程应用的前景。
