The effect of ibuprofen on the gene expression of bone differentiation markers and bcl-2 in osteoblasts

Abstract

Nonsteroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen and indomethacin inhibit prostaglandin E2 production, are commonly prescribed to reduce inflammation and pain, and slow bone loss in naturally occurring periodontal disease in beagle dogs and humans. In this study, we investigated the effects of ibuprofen on the proliferation, mRNA levels related to bone differentiation and pro-survival protein, b cell lymphoma-2 (bcl-2), of osteoblastic cells (MG63). For the this study, a human osteogenic sarcoma cell line (MG63) was used for proliferation rate (MTT assay) and total RNA extraction. The mRNA levels of alkaline phosphatase (ALP), collagen type I (COL-1), osteopontin (OPN), osteocalcin (OC), and Bcl-2 were evaluated in cells cultured with various doses of ibuprofen (in doses of 0.03 mM, 0.06 mM, 0.08 mM, 1 mM, 3.5 mM, 7 mM, 10 mM) using reverse transcription polymerase chain reaction (RT-PCR) analysis.

The results were as follows;

1. At high concentration from 7 mM to 1 mM of ibuprofen, cell proliferation rate was decreased. But, at low concentration from 0.039 mM to 0.625 mM of ibuprofen, it was about 6~40% increased. 2. The effects of ibuprofen on COL-1 mRNA in MG63 cells showed a little decreased tendency at all concentrations of ibuprofen except 3.5 mM ibuprofen compared to control. 3. The effects of ibuprofen on ALP mRNA in MG63 cells showed the slightly increased tendency at all concentrations of ibuprofen compared to control. 4. The effects of ibuprofen on OC mRNA in MG63 cells revealed the slightly increased tendency at all concentrations of ibuprofen except 7.0 mM ibuprofen compared to control. 5. The effects of ibuprofen on OPN mRNA in MG63 cells revealed dose- dependently decreased tendency. Especially, the value of OPN mRNA in 7 mM was 4 times as high as control. 6. The effects of ibuprofen on bcl-2 mRNA in MG63 cells revealed the decreased tendency at all the concentrations of ibuprofen compared to control.

Taken altogether, the local regulatory factors (ALP, especially OPN, and OC) produced by ibuprofen-treated cells were greater than those of the control, in certain concentrations of ibuprofen. Also, anti-apoptosis related gene, such as Bcl-2 was down-regulated by Ibuprofen. Further studies will be needed in relation to periodontal regeneration and anti-apoptosis at various doses (from 0.625 mM to 7 mM) of ibuprofen.