Qualitative and quantitative expression of bovine bone mineral in experimental bone defects. Part 1: Description of a dog model and histological observations

Background: The purpose of this study was to evaluate histologically the contribution of inorganic bovine bone biomaterial in a new experimental bone defect in dogs at different healing periods and to examine newly formed bone around the grafted mineral particles and their relationship in membrane-protected (test) and non-protected intrabony (control) defects. Methods: Four round intrabony defects, 5 x 4 mm were made bilaterally (at different times) on the lateral bony mandibular angle in eight dogs. Two defects were filled with bovine bone mineral (BBM) particles and two remained non-grafted but were blood clotted. A Collagen membrane covered each defect type (n = 4). This procedure was repeated on the contralateral side at a different given time to obtain two different healing periods in each dog. Thus, four specimens were obtained at 3, 6, 12, and 24 months post-operatively for each healing period. The non-decalcification method (Donath technique) with Stevenel's blue and van Gieson's picro fuchsin staining was used for histological examination. Results: Newly formed bone was observed at all examined defect types. The BBM particles were clearly evident regardless of the healing period. At 3 and 6 months, newly formed bone, woven in nature, was incorporated with the grafted particles. High cellular bone with occasional osteoclasts was noted towards the surface of the mineral particles. No substantial difference was observed between the protected and the non-protected defects except for higher ossified centers around the membrane-protected defects. At the non-grafted sites, the membrane-protected defect showed newly formed bone near the bony walls, and particularly under the membrane, establishing a bony bridge over the defect at the healing periods. The non-grafted unprotected defect (control) showed bone formation only at the base and close to the bony walls leaving a healed concave configuration. At 1 and 2 years, the grafted sites showed full bone healing configuration. However, the grafted particles still dominated the previous defect area and were completely surrounded by the newly formed bone. Osteons and lamellar bone arrangement were established but the bone was still highly cellular and osteoclasts could still be identified. The non-grafted membrane-protected sites showed excellent bone healing although areas of non-mineralized soft tissue were often seen. The control sites healed but still presented with a concave surface configuration. Conclusions: BBM biornaterial is a highly osteoconductive material. In a 4-wall bony defect, newly formed bone was well evident in establishing excellent bone healing configuration with or without a regenerative biological barrier. The grafted material dominated the experimental sites with no substantial resorption at any healing period up to 2 years observation.