A study of the influence of polysaccharides on collagen self-assembly: nanostructure and kinetics

Collagen, a critical part of the extra-cellular matrix of tissues, is a popular native material for building scaffolding for tissue-engineering applications. To mimic the structural and functional profiles of materials found in the native extra-cellular matrix, numerous efforts have been made toward developing a novel scaffold combining collagen with other biomacromolecules. All of these works have been focused on improving the mechanical or biochemical properties of the collagen-based matrix. Unfortunately, most of these studies have failed to consider the nanostructure of collagen in the complex matrix. The aim of our study was to investigate the aggregation pattern of collagen after addition of polysaccharides with positive or negative charge, the dose-response relationship, and the effect on reconstitution kinetics. Generally, collagen self-assembles into fibrils with a diameter of around 95 nm but, in the presence of various polysaccharides in varying amounts, collagen self-assembles into different shapes with larger diameters compared with collagen alone. Although the morphology and diameter of the collagen fibrils varies with reconstitution conditions, the D-periods of the fibrils all remained the same regardless of the species or concentration of polysaccharides. The kinetics of fibril formation was determined from turbidity-time curves. All turbidity curves demonstrated that polysaccharides only alter the lag time and time frame of reconstitution, but have no significant effect on the mechanism of reconstitution. Together our data indicate that the presence of biomacromolecules can alter the kinetics and the 3D fibril ultrastructure of assembled collagen and that the consequent structural changes may affect cellular responses in medical applications.