Dept. of Chemistry, McGill University, 801 Sherbrooke Street West, Montreal, Quebec H3A 0B8, Canada.
Biomacromolecules. 2024 Sep 9;25(9):6038-6049. doi: 10.1021/acs.biomac.4c00709. Epub 2024 Aug 15.
Marine mussels fabricate tough collagenous fibers known as byssal threads to anchor themselves. Threads are produced individually in minutes via secretion of liquid crystalline (LC) collagenous precursors (preCols); yet the physical and chemical parameters influencing thread formation remain unclear. Here, we characterized the structural anisotropy of native and artificially induced threads using quantitative polarized light microscopy and transmission electron microscopy to elucidate spontaneous vs regulated aspects of thread assembly, discovering that preCol LC phases form aligned domains of several hundred microns, but not the cm-level alignment of native threads. We then explored the hypothesized roles of mechanical shear, pH, and metal ions on thread formation through assembly studies employing a microfluidic flow focusing device using purified preCol secretory vesicles. Our results provide clear evidence for the role of all three parameters in modulating the structure and properties of the final product with relevance for fabrication of collagenous scaffolds for tissue engineering applications.
海洋贻贝制造出坚韧的胶原纤维,称为足丝,用于固定自身。这些纤维是通过几分钟内分泌液晶(LC)胶原前体(preCols)而单独产生的;然而,影响纤维形成的物理和化学参数仍不清楚。在这里,我们使用定量偏振光显微镜和透射电子显微镜来描述天然和人工诱导纤维的结构各向异性,以阐明纤维组装的自发和调节方面,发现 preCol LC 相形成了几百微米的定向域,但不是天然纤维的厘米级排列。然后,我们通过使用微流控流动聚焦装置进行组装研究,探索了机械剪切、pH 值和金属离子对纤维形成的假设作用,该装置使用纯化的 preCol 分泌小泡。我们的结果为这三个参数在调节最终产品的结构和性能方面的作用提供了明确的证据,这对于用于组织工程应用的胶原支架的制造具有重要意义。