Preparation and in vivo evaluation of apatite/collagen packed composite by alternate immersion method and Newton press

Further development of bio-compatible, bio-absorbable, and osteo-conductive biomaterials is desired for bone grafts in dental and medical clinics. One candidate material might be a high-density apatite/collagen composite, which cures relatively large bone defects. To produce such a composite, we freeze-dried type I collagen solution, cross-linked the formed sponge by 2 wt % glutaraldehyde, immersed the insoluble sponge in CaCl₂ and Na ₂HPO₄ solutions alternately five times, and compacted the sponge by Newton press at 5000 kgf. For comparison, cross-linked collagen without alternate immersion was also pressed. SEM/EPMA, XRD, and FTIR analyses clarified that alternate immersion successfully coated the collagen sponge with hydroxyapatite. Packed apatite/collagen composite and collagen disks 6 mm in diameter and 0.5 mm in height were implanted in the subperiostea of rabbit tibiae for 2, 4, 8, and 12 weeks to assess bio-compatibility, bio-absorbability, and osteo-conductivity. Histological observations showed that the packed apatite/collagen composite was biocompatible, osteo-conductive for up to 8 weeks, and largely bio-absorbed at 12 weeks, while the packed collagen sponge caused an undesirable foreign body reaction, which worsened with the implantation period. The overall findings suggest that this packed apatite/collagen composite might be used as a new bio-absorbable bone graft material.