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 in vitro 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.