Osteoblastic potential of infrapatellar fat pad-derived mesenchymal stem cells from rheumatoid arthritis and osteoarthritis patients

AimTo evaluate the osteoblastic potential of adipose-derived mesenchymal stem cells (ASCs) from infrapatellar fat pad (IPFP) of rheumatoid arthritis (RA) patients in comparison to osteoarthritis (OA) patients, as well as the influence of tumor necrosis factor alpha (TNF) on osteoblastic ASC differentiation in vitro. MethodsASCs were isolated from IPFP of RA and OA patients. After expansion, cells were cultured in osteogenic medium with or without TNF. After 2weeks, expression of BMP-2, Runx-2, osterix (Osx), collagen 1a1 (Col1a1) and osteopontin (OPN) messenger RNA (mRNA) was assessed by reverse transcription polymerase chain reaction and calcium deposition by alizarin red staining. Dickkopf-1 (DKK-1) and osteoprotegerin (OPG) protein concentrations were measured in culture supernatants using enzyme-linked immunosorbent assay. ResultsBoth RA- and OA-ASCs cultured in osteogenic medium showed calcium deposition. The expression of Runx2 and OPN mRNA was increased in RA-ASCs. These cells expressed significantly more Osx and OPN than OA-ASCs. TNF potentiated calcium deposition, up-regulated Runx2 and BMP-2 but down-regulated Col1a1 and OPN expression. In osteogenic cultures DKK-1 concentration was increased but that of OPG decreased, whereas TNF elevated secretion of both cytokines. ConclusionRA-ASCs have comparable or slightly stronger osteogenic potential than OA-ASCs. RA-ASCs seem to be more sensitive to TNF treatment. TNF exerts complex effects on ASC osteoblastogenesis, enhances expression of early osteogenic markers and calcium deposition, inhibits expression of mRNA coding for non-mineral bone components and alters ASC secretory activity.