Biomaterials and Tissue Engineering Laboratory, Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, Assam 781039, India.
J Tissue Eng Regen Med. 2009 Oct;3(7):539-52. doi: 10.1002/term.196.
The quest for novel materials as scaffolds with suitable micro-architecture for supporting tissue neogenesis in tissue engineering and regenerative medicine (TERM) is continuing. In this paper we report an Antheraea assama silk-based non-woven fibroin scaffold for applications in TERM. The novel three-dimensional scaffold is highly interconnected and porous, with a pore size of 150 microm, porosity of 90% and water uptake capacity of 85%. FTIR revealed a typical beta-sheet structure of fibroin. The scaffold has thermal and mechanical properties superior to those of Bombyx mori, as revealed by DSC, TGA and tensile tests. The scaffold exhibited satisfactory blood compatibility, as determined by thrombogenicity, haemolysis, platelet/leukocyte count, platelet adhesion and protein adsorption studies. The scaffold was found to be cytocompatible with human cell lines A549, KB, HepG2 and HeLa for a period of up to 4 weeks. SEM analysis revealed excellent attachment, spreading and migration of cells in the scaffold. MTT assay was performed to estimate the viability and growth of cells in the matrix. Quantification of collagen in cell-scaffold constructs was done by picro-Sirius red assay. Ex ovo chorioallantoic membrane assay and nitric oxide estimations in spent culture medium showed the scaffold's ability to promote angiogenesis. Finally, the biodegradability of the scaffold was determined by the weight loss observed upon treatment with trypsin over a period of 4 weeks. The results reveal that the fibroin from A. assama is a promising candidate as a biocompatible, biomimetic and biodegradable biomaterial of natural origin for applications in TERM.
人们一直在探索新型材料,希望找到具有合适微观结构的支架,以支持组织工程和再生医学(TERM)中的组织新生。在本文中,我们报告了一种基于柞蚕丝的无纺丝素支架,可应用于 TERM。这种新型三维支架具有高度互联和多孔的特点,孔径为 150 微米,孔隙率为 90%,吸水率为 85%。傅里叶变换红外光谱(FTIR)显示丝素具有典型的β-折叠结构。通过差示扫描量热法(DSC)、热重分析(TGA)和拉伸试验发现,支架的热学和力学性能优于桑蚕丝。通过血栓形成、溶血、血小板/白细胞计数、血小板黏附和蛋白质吸附研究,评估了支架的血液相容性,结果表明其具有良好的血液相容性。支架与人肺癌细胞系 A549、人舌鳞癌细胞系 KB、人肝癌细胞系 HepG2 和人宫颈癌细胞系 HeLa 共培养 4 周,结果表明其具有良好的细胞相容性。扫描电子显微镜(SEM)分析表明,细胞在支架中具有良好的黏附、伸展和迁移能力。MTT 试验用于评估细胞在基质中的活力和生长情况。通过苦味酸-天狼猩红染色法定量检测细胞-支架构建物中的胶原蛋白。鸡胚绒毛尿囊膜(CAM)实验和细胞培养上清液中一氧化氮(NO)的测定表明,支架具有促进血管生成的能力。最后,通过在 4 周内用胰蛋白酶处理支架观察到的重量损失来确定支架的生物降解性。结果表明,来自柞蚕的丝素是一种很有前途的生物相容性、仿生和可生物降解的天然生物材料,可应用于 TERM。