Abstract
Hydrogels made of various materials using a variety of methods have been extensively studied for use in tissue engineering, and collagen is one of the most common material used for its biocompatibility due to it being a major component of the extracellular matrix (ECM). Furthermore, the alignment of collagen fibres has been shown to direct the growth of neurites, an important criterion for engineering nervous tissues. The Multi-channel Collagen Gel (MCCG) has collagen fibres aligned circumferentially around the channel structures of the gel, and we predicted that the MCCG could guide the growth direction of neurites. In this study, we showed that the growth pathway of the neurite-like processes of PC12 cells were guided in MCCG but not in normal collagen gel (COL). The gelation of collagen gels are known to be affected by ionic concentrations, and hence we also investigated the effects of different concentrations of NaCl on the properties of MCCG. We found that, despite differences in channel density, spacing between channels, and degree of collagen fibre alignment, all MCCGs had similar guiding properties on the growth of neurites. Therefore, we believe that anisotropic MCCG could be a useful biomaterial for neural tissue engineering in the future.
Highlights
Of these, collagen is often used in scaffold construction as it is a major component of the extracellular matrix (ECM), and can satisfy many of the above-mentioned criteria
It is thought that the phase separation of collagen solution into a concentrated and diluted region results in the collagen fibres being aligned in an ordered manner parallel to the interface between these two regions[13]
From low magnification images of the top view of the Multi-channel Collagen Gel (MCCG), the neurites were seen to elongate parallel to and along the channel surface when it is close to the surface, whereas from the side view, the neurites appear to extend perpendicular to the channel structure and along the path of collgen fibre alignment (Supplementary Fig. S1)
Summary
Collagen is often used in scaffold construction as it is a major component of the ECM, and can satisfy many of the above-mentioned criteria. In other words, when the MCCG is viewed from the top, the fibres appear aligned parallel to the edges of the channel lumen, and viewed from the side, the fibres are parallel to one another but perpendicular to the channel structure (Fig. 1b). In order to minimize this free energy, the collagen fibrils adopt a conformation of aligning parallel instead of perpendicular to the channel surface, as this reduces the surface area and surface free energy of the system. We postulated that this alignment of collagen fibres may play a role in guiding the growth pathway of extending neurites. To create channel structures in COL to enable the seeding of cells, gold wires were placed in the collagen solution before gelation
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