Osteoblastic potential of infrapatellar fat pad-derived mesenchymal stem cells from rheumatoid arthritis and osteoarthritis patients.
OBJECTIVE
To 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 (TNFα) on osteoblastic ASC differentiation in vitro.


METHODS
ASCs were isolated from IPFP of RA and OA patients. After expansion, cells were cultured in osteogenic medium with or without TNFα. After 2 weeks, 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.


RESULTS
Both 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. TNFα 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 TNFα elevated secretion of both cytokines.


CONCLUSIONS
RA-ASCs have comparable or slightly stronger osteogenic potential than OA-ASCs. RA-ASCs seem to be more sensitive to TNFα treatment. TNFα 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.
Department of Pathophysiology and Immunology, Institute of Rheumatology, Warsaw, Poland.
Urszula
U
Skalska
Department of Diagnostic Haematology, Institute of Haematology and Transfusion Medicine, Warsaw, Poland.
Monika
M
Prochorec-Sobieszek
Department of Pathophysiology and Immunology, Institute of Rheumatology, Warsaw, Poland.
Ewa
E
Kontny
D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D000328:Adult; D000368:Aged; D001172:Arthritis, Rheumatoid / Q000378:metabolism* / Q000473:pathology; D015415:Biomarkers / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D020370:Osteoarthritis, Knee / Q000378:metabolism* / Q000473:pathology; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D010012:Osteogenesis / Q000187:drug effects; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D015415:Biomarkers; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D002118:Calcium
TNFα; adipose-derived mesenchymal stem cells; infrapatellar fat pad; osteoarthritis; osteoblastogenesis; rheumatoid arthritis