Dose-dependent effects of strontium on osteoblast function and mineralization

Abstract

Strontium-ranelate is now being used in the treatment of osteoporosis in elderly patients. As the majority of these patients already have a decreased renal function they are at an increased risk for accumulation of the element. Recent findings from epidemiologic studies in dialysis patients and experimental data obtained in a chronic renal failure (CRF) rat model established a dose-related multiphasic effect of strontium (Sr) on bone formation. To confirm these in vivo findings an in vitro set-up, consisting of primary rat osteoblast cultures, was applied. Sr was added to the culture medium at concentrations of 0, 0.5, 1.0, 2.0, 5.0, 20, and 100 microg/mL, respectively. Calcium incorporation (index of mineralization) and alkaline phosphatase activity were determined in the medium during the culture period, while at the end of the experiment, nodule formation (mineralized + unmineralized area) was quantified using a digital imaging system. mRNA synthesis of various osteoblast specific genes was assessed by means of reverse transcription polymerase chain reaction (RT-PCR). Compared to the control group (0 microg/mL Sr), a significantly reduced nodule formation in the presence of an intact mineralization was found for the lowest 0.5 and 1 microg/mL Sr doses, suggesting an impaired in vitro osteoblast differentiation. Both nodule formation and mineralization were normal for the 2 and 5 microg/mL doses. For the highest Sr doses (20 and 100 microg/mL) a reduced mineralization was observed in the presence of an intact nodule formation indicating an inhibitory effect on the hydroxyapatite formation. The alkaline phosphatase activity reflected the multiphasic pattern of the nodule formation while the calcium incorporation corresponded with the pattern of nodular mineralization. No variations in cell proliferation were found. RT-PCR revealed that Sr interfered with the osteoblast at the level of the mRNA synthesis of several relevant genes. Using the proposed in vitro model we confirmed the multiphasic effect of Sr on bone formation previously demonstrated in a CRF rat model. The data presented allow us to suggest that at low concentrations Sr interferes with the bone formation at the level of cell differentiation, whereas at high concentrations the disturbed mineralization in the presence of an intact nodule formation is indicative for a physicochemical interference of Sr with the hydroxyapatite formation.