Fig. 7.

Additive effect of fresh bone chips and BCM-coated DBBM on the Col1a1 and Runx2 gene expression in bone-related cell cultures. a Schematic representation of the four experimental conditions, which are compared in (b). ST2, MC3T3-E1, and primary bone–derived cells are grown for 1, 3, and 7 days either on (1) BCM-free DBBM granules hydrated with RS + S, a condition labeled as control (Ctrl) or (2) DBBM granules coated with BCM prepared within 20 min in RS + S, a condition labeled as BCM. Experimental conditions (3) bone chips (BCh) and (4) BCM/BCh duplicate conditions (1) and (2) in the presence of fresh bone chips placed in a cell culture insert with a 0.4-μm pore size. b, c Effect of BCM-coated DBBM in the absence or presence of fresh bone chips on Col1a1 (b) and Runx2 (c) mRNA expression levels in ST2, MC3T3-E1, and primary bone–derived cells. Cells were grown under each of the four experimental conditions described in (a) for 1, 3, and 7 days before total RNA was isolated and analyzed by qRT-PCR. Values normalized to Gapdh are expressed relative to the values of control cells at the “1 day” time point. Please note the differences in the scales of the y-axis. Data represent means ± SD from three independent experiments. Significant differences to the respective controls at day 1 unless otherwise indicated, ***P < 0.001, **P < 0.01, *P < 0.05