Abbm c

The experimental mAb was a chimeric anti-BMP-2 mAb with cross-reactivity to BMP-4 and BMP-7. The control mAb was an isotype matched mAb specific for the KLH peptide that had no specific affinity for BMP-2 [54 (link)]. A concentration of 25 μg/ml of mAb was chosen based on the results of our previous studies [54 (link)]. Anti-BMP-2 mAb and isotype matched control mAb were immobilized on deproteinized anorganic bovine bone mineral with 10% collagen (ABBM-C; Bio-Oss Collagen®, Geistlich, Pharma AG, Wolhusen, Switzerland) as well as porcine bilayer native collagen membrane (CM; Bio-Gide® membrane, Geistlich, Pharma AG, Wolhusen, Switzerland) as previously described [54 (link)]. Briefly, the ABBM-C and CM were incubated at room temperature with mAb diluted with phosphate-buffered saline (PBS) for one hour prior to implantation into the sockets after tooth extraction. All antibody preparations were made by two of the coauthors (S.M., O.K.).

Anorganic bovine bone mineral with 10% collagen (ABBM-C; Bio-Oss-Collagen, Geistlich Pharma, Wolhusen, Switzerland) was used as the scaffold in AMOR. ABBM has interconnected macropores to allow for neovascularization and bone cell infiltration and micropores to allow for fluid exchange, as well as nanostructure to help with osteogenesis [50 (link)]. Chimeric anti-BMP-2 mAb recently developed in our laboratory [47 (link)] was used in this study. The negative control consisted of isotype-matched mAb (Iso, anti-rabbit IgG2a mAb, Biovision, Mountain View, CA) with no specific affinity to BMP-2. Anti-BMP-2 mAb and its isotype-matched control mAb were diluted with phosphate-buffered saline (PBS) at 25 μg/mL concentration. Recombinant human BMP-2 (Infuse, Medtronic, Inc., Memphis, TN, USA) was used at 20 μg/ml as positive control. Scaffolds were incubated with 250 μl of diluted anti-BMP-2 mAb (total of 6.25 μg), isotype-matched mAb (total of 6.25 μg), or rhBMP-2 (total of 5.0 μg), as previously described [42 (link)].