Osteogenesis of bone marrow stromal cell in DBP sponge with various contents

Demineralized bone particle(DBP) that affects to cell proliferation and differentiation has been used as biomaterials. Bone marrow stromal cells(BMSCs) exhibit multiple traits of a stem cell population, and they can expand many times in vitro and be induced to differentiate into multiple cell types. In this study, we evaluated the effect of type of medium and contents DBP sponge on bone differentiation of rat BMSCs. Growth medium was consisted of DMEM, 10% FBS, 1% antibiotic antinycotics and Osteogenesis medium added to β-glycerophosphate 10 mM, L-ascorbic acid, dexamethasone 10^-8 M, vitamin D₃ 10^-8 M. DBP sponge were prepared by freeze-drying method from 1, 2 and 3 wt% DBP solution. BMSCs were harvested from the femurs of adult female Fischer rat. The effect of DBP sponge on the proliferation and ostegenesis of rat BMSCs were assessed in culture using the MTT assay, SEM, ALP assay, and RT-PCR was conducted to confirm mRNA expression of ALP and osteocalcin for osteogenic marker. According to our results, the cell viability and the quantity of ALP in 3 wt% DBP sponge were superior to other sponges. The osteogenesis medium provided the better osteogenesis effect than the growth medium. We concluded that osteogenesis medium and 3 wt% DBP sponge will be useful to bone differentiation using 3 wt% DBP sponge.