Growth of new bone guided by implants in a murine calvarial model

Abstract

New methods to increase vertical bone growth are needed to permit dental implant placement in patients with low alveolar ridge height after extended periods of tooth loss. While ectopic rodent models are typically used to evaluate new osteogenic implant surface coatings, a more relevant intramembraneous rodent model was needed to address the particular clinical need to grow a new layer of bone above an existing layer of bone. In this study we report on a novel murine calvaria model in which successful vertical bone growth around miniaturized dental implants was achieved when using non-glycosylated bone morphogenetic protein-2 (ng/rhBMP-2). Twenty CD-1 mice received two Ti implants each consisting of a Ti ring implant stabilized by a Ti screw into the occipital calvarial bone. Four groups were evaluated: control Ti, Ti+20 mug ng/rhBMP-2, hydroxyapatite (HA)-coated Ti, and HA+20 mug ng/rhBMP-2. The mice were sacrificed 21 days following implant placement. MicroCT analysis showed no new bone formation around the untreated Ti or the HA-coated implants, but demonstrated new bone growth in every dimension around and above the Ti+ng/rhBMP-2 and the HA+ng/rhBMP-2 treated implants. Histopathologic analysis showed that a thin fibrous capsule covered the untreated Ti implants. Limited bone-to-implant contact (BIC) was observed for the HA-coated implants, while in contrast both ng/rhBMP-2 treated groups exhibited extensive new supracalvarial woven bone that covered the implant and merged with the calvarial plate. Histomorphometrically, supracalvarial bone heights and bone widths and BIC were not statistically different from one another for the two ng/rhBMP-2 treated groups. However, the total supracalvarial bone surface area was significantly greater (p<0.05) for the Ti+ng/rhBMP-2 implants (7.2 mm(2)) than the HA+ng/rhBMP-2 (4.0 mm(2)) treated implants. The bone density within 1 mm around the implant was also significantly greater (p<0.05) for the Ti+ng/rhBMP-2 implants (9.9%) than the HA+ng/rhBMP-2 (4.0%) implants, indicating that HA coatings may not be required for sustained release when non-glycosylated BMP-2 is used. This new murine model is capable of discriminating between various bone augmentation strategies and may represent a clinically more relevant model for alveolar bone augmentation than the commonly used ectopic muscle pouch or long bone models.