Effects of material and surface functional group on collagen self-assembly and subsequent cell adhesion behaviors

Collagen fibrous network not only provides structural support for cells but also serves as critical environment modulating various cell functions. Various factors would influence the collagen self-assembly but the effect of substrate surface on such process has been rarely studied. Here we examined the effects of materials (Ti and hydroxyapatite) and their surface characteristics (with and without the enrichment of hydroxyl group) on collagen self-reconstitution and fibrous network formation, and on subsequent cell adhesion and cytoskeleton organization of mesenchymal stem cells (MSCs). For both Ti and hydroxyapatite (HA) substrates, the enrichment of hydroxyl group (OH) on substrate surfaces promoted the collagen self-reconstitution and facilitated the formation of the fibrous network after 4h immersion in phosphate buffer solution (PBS), while all samples showed clear fibrous network formation after 2 day soaking in PBS. Compared with the Ti surfaces, the HA surfaces facilitated the self-reconstitution of collagen, leading to a more mature fibrous network with a twisted structure and enhanced lateral aggregation of fibrils. The fibrous network difference resulted in different behaviors of the subsequent MSC adhesion and spreading. The MSCs had the best adhesion and cytoskeleton organization on the OH enriched HA surface with collagen modification. Our results suggested that both the material selection and the hydroxyl group significantly influenced the collagen self-assembly and fibrous network formation and, as a result, the subsequent cell adhesion behaviors.