Evaluation of the Osteoinductive Capacity of Canine Demineralized Bone Matrix in Heterotopic Muscle Sites of Athymic Rats

Abstract

SummaryThe osteoinductive capacity of canine demineralized bone matrix (DBM), implanted in epaxial muscle sites of athymic rats, was evaluated using calcium content and histomorphometry at two, four and six weeks after implantation. Results of this study confirm that DBM, derived from canine sources, does possess significant osteoinductive ability since histological examination revealed the presence of new cartilage, bone, or both, at 21/24 implantation sites. The osteogenesis induced by canine DBM continued as an active, cumulative process throughout the six week investigation period. The mean percentage of total induced osteogenic components including new, live cartilage, woven bone, lamellar bone and bone marrow cellular elements, was significantly greater after six weeks than after two weeks of implantation (p <0.01). Comparison of histomorphometric point counts at two, four and six weeks of implantation supported the conclusion that bone for mation as induced by canine DBM, proceeds primarily via an endochondral ossification pathway. Although the amount of calcium deposited in tissues harvested from DBM implanted sites tended to increase as implantation time lengthened, there was not a statistically significant correlation between calcium content and the level of osteogenic activity seen histologically (r = 0.32, p = 0.13).The osteoinductive capacity of canine demineralized bone matrix (DBM), implanted in ep-axial muscle sites of athymic rats, was evaluated using calcium content and histomorphometry at two, four and six weeks after implantation. Results of this investigation confirm that DBM, derived from canine sources, does possess significant osteo-inductive ability and that bone formation proceeds primarily via a pathway of endochondral ossification.

Comparison of Bone Healing by Demineralized Bone Matrix and Autogenous Cancellous Bone in Horses

“…Results from these studies confirm a particle size effect on osteoinduction, and also describe different responses to intramuscular and subcutaneous implantation 24,26 . Intermediate formation of cartilage is also much less marked in horses than it is in rats implanted with DBM 26,27 . No reports describe the reaction of DBM in orthotopic sites in horses.…”

“…Also, random samples of DBM were dried at 95°C, weighed, and then ashed at 600°C for 24 hours. These samples were then dissolved in 0.6 N nitric acid and analyzed by atomic absorption spectrophotometry to determine percent calcium per gram dry weight (% Ca:DW) 27,28 . Demineralization was considered adequate when samples were no longer visible radiographically and when calcium content was less than 1% 29 …”

“…Each rib was then sectioned in the transverse plane at the middle of each defect, and a 4‐mm wide, full‐thickness section was obtained dorsal and ventral to the cut. The section dorsal to the cut was sampled for calcium and ash content by atomic absorption spectrophotometry (described earlier), and the percent calcium and percent ash per dry weight were determined for each defect 27,28 . The remaining section ventral to the cut was decalcified in 10% formic acid with constant agitation over ion resin exchange, dehydrated in alcohol, embedded in paraffin, sectioned at 7 μm, and stained with hematoxylin and eosin.…”

Evaluation of bone healing in canine tibial defects filled with cortical autograft, commercial-DBM, calf fetal DBM, omentum and omentum-calf fetal DBM

“…Loss of density radiographically was used to subjectively evaluate demineralization. In addition, random samples of DBM were dried at 95℃, weighed, and then ashed at 600℃ for 24 h. These samples were subsequently dissolved in 0.6 mol/L nitric acid and analyzed by atomic absorption spectrophotometry to determine the percent calcium per gram dry weight (% Ca: DW) [12,29]. Demineralization was considered adequate when samples were no longer visible radiographically and when the calcium content was less than 1% [38].…”

Experimental bone defect healing with xenogenic demineralized bone matrix and bovine fetal growth plate as a new xenograft: radiological, histopathological and biomechanical evaluation

“…Loss of density radiographically was used to subjectively evaluated demineralization. Also, random samples of DBM were dried at 95°C, weighed, and then ashed at 600°C for 24 h. These samples were then dissolved in 0.6 mol/l nitric acid and analyzed by atomic absorption spectrophotometry to determine percent calcium per gram dry weight (% Ca:DW) (Reddi and Huggins 1972;Forell et al 1993). Demineralization was considered adequate when samples were no longer visible radiographically and when calcium content was less than 1% (Urist and Strakes 1970).…”