title: Compensatory cellular reactions to nonsteroidal anti-inflammatory drugs on osteogenic differentiation in canine bone marrow-derived mesenchymal stem cells.
abstract: The suppressive effects of nonsteroidal anti-inflammatory drugs (NSAIDs) on the bone healing process have remained controversial, since no clinical data have clearly shown the relationship between NSAIDs and bone healing. The aim of this study was to assess the compensatory response of canine bone marrow-derived mesenchymal stem cells (BMSCs) to several classes of NSAIDs, including carprofen, meloxicam, indomethacin and robenacoxib, on osteogenic differentiation. Each of the NSAIDs (10 µM) was administered during 20 days of the osteogenic process with human recombinant IL-1β (1 ng/ml) as an inflammatory stimulator. Gene expression of osteoblast differentiation markers (alkaline phosphatase and osteocalcin), receptors of PGE2 (EP2 and EP4) and enzymes for prostaglandin (PG) E2 synthesis (COX-1, COX-2, cPGES and mPGES-1) was measured by using quantitative reverse transcription-polymerase chain reaction. Protein production levels of alkaline phosphatase, osteocalcin and PGE2 were quantified using an alkaline phosphatase activity assay, osteocalcin immunoassay and PGE2 immunoassay, respectively. Histologic analysis was performed using alkaline phosphatase staining, von Kossa staining and alizarin red staining. Alkaline phosphatase and calcium deposition were suppressed by all NSAIDs. However, osteocalcin production showed no significant suppression by NSAIDs. Gene expression levels of PGE2-related receptors and enzymes were upregulated during continuous treatment with NSAIDs, while certain channels for PGE2 synthesis were utilized differently depending on the kind of NSAIDs. These data suggest that canine BMSCs have a compensatory mechanism to restore PGE2 synthesis, which would be an intrinsic regulator to maintain differentiation of osteoblasts under NSAID treatment.
authors.affiliation: Laboratory of Veterinary Surgery, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
authors.forename: Namgil
authors.initials: N
authors.lastname: Oh
authors.forename: Sangho
authors.initials: S
authors.lastname: Kim
authors.forename: Kenji
authors.initials: K
authors.lastname: Hosoya
authors.forename: Masahiro
authors.initials: M
authors.lastname: Okumura
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002227:Carbazoles / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D017931:DNA Primers / Q000235:genetics; D015232:Dinoprostone / Q000096:biosynthesis*; D004159:Diphenylamine / Q000031:analogs & derivatives / Q000494:pharmacology; D004285:Dogs; D020869:Gene Expression Profiling / Q000662:veterinary; D005786:Gene Expression Regulation / Q000187:drug effects*; D005819:Genetic Markers / Q000235:genetics; D007118:Immunoassay / Q000662:veterinary; D007213:Indomethacin / Q000494:pharmacology; D053583:Interleukin-1beta; D000077239:Meloxicam; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects*; D010648:Phenylacetates / Q000494:pharmacology; D018709:Statistics, Nonparametric; D013843:Thiazines / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D002227:Carbazoles; D017931:DNA Primers; D005819:Genetic Markers; D053583:Interleukin-1beta; D010648:Phenylacetates; D013843:Thiazines; D013844:Thiazoles; D015675:Osteocalcin; D004159:Diphenylamine; D000469:Alkaline Phosphatase; C007005:carprofen; D015232:Dinoprostone; D002118:Calcium; D000077239:Meloxicam; D007213:Indomethacin; C551524:robenacoxib