Cells and Materials


Bone can be described as a highly ordered composite of type I collagen integrated with an inorganic mineral phase. In vitro models of bone mineralization using collagenous substrates have been reported in the literature. This study reports an in vitro system of mineralized reconstituted collagen fibers, with aligned fibrillar substructure. The collagen fibers were mineralized in a double diffusion chamber saturated with respect to calcium and phosphate. The morphology and ultrastructure of the mineral precipitate were evaluated as a function of the pH of the incubating media. Brushite crystal was observed at acidic pH. Large rectangular crystals formed at pH 5.15 and appear to associate with the collagen fibers. At neutral and alkaline pHs, hydroxyapatite crystals were observed in association with the collagen fibers . Spherical aggregates of hydroxyapatite crystals were seen at neutral and alkaline pHs, but these structures were reduced in size when formed on collagen at alkaline pH. On close examination these spherical structures were found to be hollow when viewed in cross section.

The crystals precipitated within the interior of the collagen fiber at neutral and alkaline pHs were comparable in both size and shape to crystals observed in mineralized turkey tendon and skeletal tissues. These preliminary observations indicate that with further refinement the reconstituted collagen fibers may prove useful in model systems for the study of collagen mediated mineralization in vitro. In addition, mineralization of collagenous matrices may lead to the development of biomaterials for bone repair and replacement.