The development of bone-like collagen-hydroxyapatite composites is highly desirable because bone has outstanding mechanical properties and resorptive potential, and a combination of these characteristics could ultimately lead to a load-bearing and bioresorbable bone substitute. Our prior work has shown that intrafibrillar mineralization of collagen can be achieved using a polymer-induced liquid-precursor (PILP) mineralization process. In our in vitro model system, polyaspartate is used to mimic the acidic non-collagenous proteins involved in bone formation. We have previously shown that the anionic polypeptide sequesters ions to induce an amorphous calcium phosphate precursor, and we have put forth the hypothesis that the early-stage precursor is highly hydrated, enabling fluidic droplets to be drawn into the gaps and grooves of collagen fibrils by capillary action. Here, we further our biomimetic approach by using alkaline phosphatase to provide a slow release of inorganic phosphate ions from a phosphate ester, mimicking the biochemical processes of ion regulation found in natural bone formation. The collagen-hydroxyapatite composites were characterized using transmission electron microscopy (TEM) and selected area electron diffraction (SAED), which show that nanocrystals of hydroxyapatite are intrafibrillar and [0 0 1] oriented along the collagen fibril axis. With repeated mineralization steps, the fibrils become cemented together with a non-descript extrafibrillar mineral coating. Although the degree of intrafibrillar mineralization was not as high as our usual method, we believe that with further optimization this enzyme-aided PILP process could provide a closer mimic to the biochemical processes involved in bone formation, and serve as a useful in vitro model system for studying the mechanisms involved in bone formation.
Bibliographical noteFunding Information:
This material is based upon work supported by the National Science Foundation under award No. BES-0404000 .
Copyright 2010 Elsevier B.V., All rights reserved.
- A1. Biocrystallization
- A1. Biomaterials
- A2. Growth from solutions
- B1. Calcium compounds, Hydroxyapatite, Amorphous precursor