Simvastatin and atorvastatin enhance gene expression of collagen type 1 and osteocalcin in primary human osteoblasts and MG‐63 cultures

Abstract

To clarify the mechanism of the stimulatory effect of statins on bone formation, we have assessed the effect of simvastatin and atorvastatin on osteoblast activity by analysing cell proliferation, as well as collagen, osteocalcin, and bone morphogenetic protein-2 (BMP2) gene expression in primary human osteoblast (hOB) and MG-63 cell line cultures. Explants of bone from patients without any metabolic disease under orthopedic hip procedures were used to obtain hOB. Cell cultures were established, synchronized, and different concentrations of simvastatin or atorvastatin were added (10(-9) M, 10(-8) M, 10(-7) M, 10(-6) M) during the experiment. Cell proliferation was analyzed after 24 h. Collagen polypeptide alpha1 type 1 (COL1A1) gene expression, osteocalcin, and BMP2 expression levels were quantified by real-time PCR after 24 h incubation with statins. There was a statistically significant decrease in cell proliferation related to simvastatin or atorvastatin addition at all concentrations in primary hOB compared with those not treated. A significant increase in COL1A1, osteocalcin, and BMP2 gene expression was detected when hOB cultures were treated with simvastatin or atorvastatin at different concentrations. Similar but less significant effects were found on MG-63 cells. After statin treatment we observed both an arrest of proliferation in hOB cells and an increase in collagen, osteocalcin, and BMP2 gene expression, consistent with a stimulatory effect towards mature osteoblast differentiation. These findings support the bone-forming effect of statins, probably through the BMP2 pathway.

Simvastatin inhibits cytokine‐stimulated Cyr61 expression in osteoblastic cells: A therapeutic benefit for arthritis

“…Exogenous Cyr61 enhanced the growth of both types of osteoblastic cells, whereas simvastatin suppressed the proliferation. Our findings are in accordance with those of previous studies which showed that Cyr61 enhanced the proliferation of osteoblasts (33) and statins stimulated differentiation but induced growth arrest in osteoblasts (34). Since the responses were similar in the 2 cell types and U2OS cells have more stable biologic behavior and higher transfection efficiency, they were used to study the associated signaling pathway.…”

Rosuvastatin Promotes Osteoblast Differentiation and Regulates SLCO1A1 Transporter Gene Expression in MC3T3-E1 Cells

“…RSV, similarly to our results after 24h incubation, showed absence of cytotoxicity in kidney cells in the concentration range of 0.001-100 µM [47]. In other studies in primary human osteoblasts and osteosarcoma cells, a cell viability above 85% was observed using simvastatin and atorvastatin with concentrations ranging from 0.001-1 µM [19]. In a human bladder carcinoma T24 cell line, lovastatin was found to be cytotoxic at a concentration of 50 µM [48].…”

“…The treatment with RSV had no effects on cell proliferation within the concentration range of 0.001-10 µM, and significantly reduced proliferation with 100 µM of RSV, which could be related to the higher cytotoxicity found with this dose. The antiproliferative effect of statins has been shown to be more evident in cultures of human bone marrow stromal cells [15] and primary human osteoblasts [19], while it was negligible with the MG-63 osteosarcoma cell line [19]. Therefore, the high growth rate of transformed osteoblastic cell lines could account for the lack of antiproliferative effects in these in vitro models.…”

Statins: candidates for promoting bone formation via BMP-2

“…Randomized controlled trials of the statin family are needed to confirm the risk of fractures (7). The statin family enhances the mRNA and protein expression levels of BMP-2 and vascular endothelial growth factor (VEGF) to stimulate osteoblastic functions and upregulate gene expression of extracellular matrices including osteocalcin, bone sialoprotein, collagens and proteoglycans (2,(8)(9)(10). Statins induce and accelerate bone formation locally, trigger early development of growth factors, regulate angiogenesis by VEGF, differentiate into osteoblasts or osteocytes, and cause bone mineralization (11)(12).…”