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http://dx.doi.org/10.5051/jpis.2012.42.6.224

In vitro assay for osteoinductive activity of different demineralized freeze-dried bone allograft  

Vaziri, Shahram (Department of Periodontology, Dental school, Shahid Beheshti University of Medical Sciences)
Vahabi, Surena (Department of Periodontology, Dental school, Shahid Beheshti University of Medical Sciences)
Torshabi, Maryam (Department of Dental Material, Dental school, Shahid Beheshti University of Medical Sciences)
Hematzadeh, Somayeh (Department of Periodontology, Dental school, Qazvin University of Medical Sciences)
Publication Information
Journal of Periodontal and Implant Science / v.42, no.6, 2012 , pp. 224-230 More about this Journal
Abstract
Purpose: Various bone graft materials have been used for periodontal tissue regeneration. Demineralized freeze-dried bone allograft (DFDBA) is a widely used bone substitute. The current widespread use of DFDBA is based on its potential osteoinductive ability. Due to the lack of verifiable data, the purpose of this study was to assess the osteoinductive activity of different DFDBAs in vitro. Methods: Sarcoma osteogenic (SaOS-2) cells (human osteoblast-like cells) were exposed to 8 mg/mL and 16 mg/mL concentrations of three commercial types of DFDBA: Osseo+, AlloOss, and Cenobone. The effect of these materials on cell proliferation was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The osteoinductive ability was evaluated using alizarin red staining, and the results were confirmed by evaluating osteogenic gene expression using reverse transcription polymerase chain reaction (RT-PCR). Results: In the SaOS-2 cells, an 8 mg/mL concentration of Osseo+ and Cenobone significantly increased cell proliferation in 48 hours after exposure (P<0.001); however, in these two bone materials, the proliferation of cells was significantly decreased after 48 hours of exposure with a 16 mg/mL concentration (P<0.001). The alizarin red staining results demonstrated that the 16 mg/mL concentration of all three tested DFDBA induced complete morphologic differentiation and mineralized nodule production of the SaOS-2 cells. The RT-PCR results revealed osteopontin gene expression at a 16 mg/mL concentration of all three test groups, but not at an 8 mg/mL concentration. Conclusions: These commercial types of DFDBA are capable of decreasing proliferation and increasing osteogenic differentiation of the SaOS-2 cell line and have osteoinductive activity in vitro.
Keywords
Alizarin red; Bone graft; Cell differentiation; Cell proliferation; Regeneration;
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