Chakraborty Juhi, Fernández-Pérez Julia, van Kampen Kenny A, Roy Subhadeep, Ten Brink Tim, Mota Carlos, Ghosh Sourabh, Moroni Lorenzo
Department of Textile and Fibre Engineering, Indian Institute of Technology Delhi, New Delhi 110016, India.
Department of Complex Tissue Regeneration, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, 6211 LK Maastricht, The Netherlands.
Biofabrication. 2023 Apr 14;15(3). doi: 10.1088/1758-5090/acc68f.
In recent years, engineering biomimetic cellular microenvironments have been a top priority for regenerative medicine. Collagen II, which is arranged in arches, forms the predominant fiber network in articular cartilage. Due to the shortage of suitable microfabrication techniques capable of producing 3D fibrous structures,replication of the arch-like cartilaginous tissue constitutes one of the major challenges. Hence, in the present study, we report a 3D bioprinting approach for fabricating arch-like constructs using two types of bioinks, gelatin methacryloyl (GelMa) and silk fibroin-gelatin (SF-G). The bioprinted SF-G constructs displayed increased proliferation of the encapsulated human bone marrow-derived mesenchymal stem cells compared to the GelMA constructs. Biochemical assays, gene, and protein expression exhibited the superior role of SF-G in forming the fibrous collagen network and chondrogenesis. Protein-protein interaction study using Metascape evaluated the function of the proteins involved. Further GeneMANIA and STRING analysis using Col 2A1, SOX 9, ACAN, and the genes upregulated on day 21 in RT-PCR, i.e.-catenin, TGFR1, Col 1A1 in SF-G and PRG4, Col 10A1, MMP 13 in GelMA validated ourresults. These findings emphasized the role of SF-G in regulating the Wnt/-catenin and TGF-signaling pathways. Hence, the 3D bioprinted arch-like constructs possess a substantial potential for cartilage regeneration.
近年来，工程化仿生细胞微环境一直是再生医学的首要任务。呈拱形排列的Ⅱ型胶原蛋白构成了关节软骨中的主要纤维网络。由于缺乏能够制造三维纤维结构的合适微加工技术，复制拱形软骨组织是主要挑战之一。因此，在本研究中，我们报告了一种三维生物打印方法，该方法使用两种生物墨水——甲基丙烯酰化明胶（GelMa）和丝素蛋白-明胶（SF-G）来制造拱形结构。与GelMA结构相比，生物打印的SF-G结构显示出被封装的人骨髓间充质干细胞增殖增加。生化分析、基因和蛋白质表达显示了SF-G在形成纤维状胶原网络和软骨形成方面的优越作用。使用Metascape进行的蛋白质-蛋白质相互作用研究评估了所涉及蛋白质的功能。进一步使用Col 2A1、SOX 9、ACAN以及在实时定量聚合酶链反应（RT-PCR）中第21天上调的基因，即SF-G中的β-连环蛋白、转化生长因子受体1（TGFR1）、Ⅰ型胶原蛋白α1（Col 1A1）以及GelMA中的润滑蛋白（PRG4）、Ⅹ型胶原蛋白α1（Col 10A1）、基质金属蛋白酶13（MMP 13）进行的基因共表达网络分析（GeneMANIA）和搜索工具检索感兴趣基因的字符串数据库（STRING）分析验证了我们的结果。这些发现强调了SF-G在调节Wnt/β-连环蛋白和转化生长因子信号通路中的作用。因此，三维生物打印的拱形结构在软骨再生方面具有巨大潜力。
