Collagen supramolecular and suprafibrillar organizations on osteoblasts long-term behavior: benefits for bone healing materials.

This study compares the behavior of osteoblastic cells seeded on three structurally distinct collagen-based materials. Adhesion and long-term behavior were evaluated in vitro in regard to collagen scaffolds forming loose or dense fibrillar networks or exempt of fibrils. In this purpose collagen solutions at concentrations of 5 and 40 mg/mL were processed by freeze-drying or by sol/gel fibrillogenesis to form either sponges or hydrogels. Macroscopic and microscopic images of sponges showed a light material exhibiting large pores surrounded by dense collagen walls made of thin unstriated microfibrils of 20 nm in diameter. In comparison collagen hydrogels are more homogeneous materials, at 5 mg/mL the material consists of a regular network of cross-striated collagen fibrils of 100 nm in diameter. At 40 mg/mL the material appears stiffer, the ultrastructure exhibits cross-striated collagen fibrils packed in large bundles of 300-800 nm of width. Human osteoblastic cells seeded on top of the 5 mg/mL matrices exhibit a squared shaped osteoblast-like morphology over 28 days of culture and express both alkaline phosphatase and osteocalcin. Osteoblastic cells seeded on top of sponges or of 40 mg/mL matrices exhibit both flat and elongated resting-osteoblast morphology. Osteoblastic cells have mineralized the three collagen-based materials after 28 days of culture but collagen sponges spontaneously mineralized in absence of cells. These results highlight, in an in vitro cell culture approach, the benefit of fibrils and of dense fibrillar networks close to in vivo-like tissues, as positive criteria for new bone tissue repair materials.