Commercially-prepared allograft material has biological activity in vitro.
The well-established finding that implantation of demineralized bone matrix at non-skeletal sites results in formation of cartilage and bone has been attributed to bone morphogenetic proteins/factors. Commercially-available demineralized bone allograft materials are being used currently to reconstruct/regenerate bone. The studies described here focused on establishing biological activity of protein extracts prepared from commercially obtained bone graft material in vitro. Furthermore, the biological activity of these protein extracts in vitro was compared with similar extracts prepared from freshly obtained human bone. Biological activities of bone matrix proteins examined included their ability to promote proliferation, attachment, and migration of gingival fibroblasts using an in vitro system. Guanidine followed by guanidine/EDTA was used to separate bone matrix proteins into proteins associated with soft tissues of bone and proteins retained within the mineral compartment, respectively. Two preparations of each starting material were tested and the biological activity of each preparation was evaluated in triplicate at least three times. Slot blot analysis revealed that commercially-prepared material contained type I collagen; fibronectin; BSP; and BMP-2, 4, and 7. However, the freshly prepared bone extracts appeared to have higher BMP concentrations. The ability of commercial extracts to promote cell proliferation, while significant, was limited and significantly less when compared with similar extracts prepared from freshly obtained bone. All extracts promoted cell attachment significantly, while none of the extracts promoted cell migration. Thus, commercially-prepared material retained proteins having the capacity to influence cell behavior in vivo. However, some biological activity as measured in vitro was lost as a result of tissue processing.