Bone+B bone xenograft is an inorganic bone matrix produced through eliminating organic compounds from the bovine cancellous bone. For this purpose, using accurate scientific methods, the bovine bone is subject to chemical and thermal treatments so that all of its organic content including proteins, DNA, blood and fat cells are removed while the crystal and porous structure of hydroxyapatite is preserved. Scanning electron microscopy (SEM) of the bone powder confirms the porous structure of this compound.
Bone+B is designed and produced for the following applications:
To investigate the genotoxic effects of Bone+B using the comet assay method, normal saline was added to bone powder and the mixture was kept at 37°C for one week. The filtered extract was used to treat L929 cells. For this purpose, the L929 cells were cultured in a 24-well plate and treated with the Bone+B extract. Normal saline and hydrogen peroxide was used as negative and control, respectively. After incubating the plates at 37°C for 3 hours, 50 µl of each sample was mixed with 50 µl dissolved agarose and the resultant suspension was spread on special comet assay slides. The slides were irrigated with 1XTBE twice and then electrophoresed for 10 minutes. Finally, the slides were visualized under a fluorescent microscope after fixation, drying, and staining with ethidium bromide.
The comet assay was done by measuring the mean DNA head and tail length after treatment of cells with Bone+B Extract. The results were analyzed statistically using unpaired students’t test, one-way ANOVA, and Dunnest’s post hoc test. The Bone+B extract caused no significant changes compared to the control group; thus, the results of the comet assay was negative.
According to ISO 10993-3, Bone+B lacks genotoxic effects.