BMP-4, TGF-β and Smad3 as Modulators of Synovial Fluid Cells Viability^[*]

Abstract

Objective Our goal was to evaluate the modulation of the synovial fluid cells (SFC) from patients with and without osteoarthritis (OA) by bone morphogenetic protein 4 (BMP-4), Smad-3 and transforming growth factor beta (TGF-β).

Methods Synovial fluid was collected from patients submitted to knee arthroscopy or replacement and were centrifuged to isolate cells from the fluid. Cells were cultured for 21 days and characterized as mesenchymal stem cells (MSCs) according to the criteria of the International Society of Cell Therapy. Then, we performed an [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay (MTT) assay after exposing cells with and without OA to TGF-β, Smad3 and BMP-4 pathway inhibitors and to different concentrations of BMP4.

Results Exposure to the TGF-β, Smad3 and BMP-4 inhibitors modifies the mitochondrial activity of the SFCs. The activity of the SFCs is modified by influences of increasing concentrations of BMP4, but there is no difference in cellular activity between patients with and without OA.

Conclusion TGF-β, Smad3 and BMP-4 modulate the activity of SFCs from patients with and without knee OA.

Keywords

mesenchymal stem cells - osteoarthritis - synovial fluid cells - TGF-β pathway

Author Contributions

de Souza E. B.: Concept and design, provision of study material or patients, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of the manuscript.

Moura Neto V.: Data analysis and interpretation, final approval of the manuscript.

Aguiar D. P.: Concept and design, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of the manuscript.

All authors read and approved the final manuscript.

^* Work developed at the Divisão de Ensino e Pesquisa, Instituto Nacional de Ortopedia e Traumatologia Jamil Haddad, Rio de Janeiro, RJ, Brazil.

Publication History

Received: 31 May 2020

Accepted: 16 September 2020

Article published online:
31 March 2021

• Referências

• 1 Gupta PK, Das AK, Chullikana A, Majumdar AS. Mesenchymal stem cells for cartilage repair in osteoarthritis. Stem Cell Res Ther 2012; 3 (04) 25
• 2 Johnson K, Zhu S, Tremblay MS. et al. A stem cell-based approach to cartilage repair. Science 2012; 336 (6082): 717-721
• 3 Barry F, Murphy M. Mesenchymal stem cells in joint disease and repair. Nat Rev Rheumatol 2013; 9 (10) 584-594
• 4 Pers YM, Ruiz M, Noël D, Jorgensen C. Mesenchymal stem cells for the management of inflammation in osteoarthritis: state of the art and perspectives. Osteoarthritis Cartilage 2015; 23 (11) 2027-2035
• 5 Jones EA, Crawford A, English A. et al. Synovial fluid mesenchymal stem cells in health and early osteoarthritis: detection and functional evaluation at the single-cell level. Arthritis Rheum 2008; 58 (06) 1731-1740
• 6 Sekiya I, Ojima M, Suzuki S. et al. Human mesenchymal stem cells in synovial fluid increase in the knee with degenerated cartilage and osteoarthritis. J Orthop Res 2012; 30 (06) 943-949
• 7 van der Kraan PM, Goumans MJ, Blaney Davidson E, ten Dijke P. Age-dependent alteration of TGF-β signalling in osteoarthritis. Cell Tissue Res 2012; 347 (01) 257-265
• 8 Keller B, Yang T, Chen Y. et al. Interaction of TGFβ and BMP signaling pathways during chondrogenesis. PLoS One 2011; 6 (01) e16421
• 9 Mariani E, Pulsatelli L, Facchini A. Signaling pathways in cartilage repair. Int J Mol Sci 2014; 15 (05) 8667-8698
• 10 Leonard CM, Fuld HM, Frenz DA, Downie SA, Massagué J, Newman SA. Role of transforming growth factor-beta in chondrogenic pattern formation in the embryonic limb: stimulation of mesenchymal condensation and fibronectin gene expression by exogenenous TGF-beta and evidence for endogenous TGF-beta-like activity. Dev Biol 1991; 145 (01) 99-109
• 11 Song JJ, Aswad R, Kanaan RA. et al. Connective tissue growth factor (CTGF) acts as a downstream mediator of TGF-beta1 to induce mesenchymal cell condensation. J Cell Physiol 2007; 210 (02) 398-410
• 12 van der Kraan PM, Blaney Davidson EN, Blom A, van den Berg WB. TGF-beta signaling in chondrocyte terminal differentiation and osteoarthritis: modulation and integration of signaling pathways through receptor-Smads. Osteoarthritis Cartilage 2009; 17 (12) 1539-1545
• 13 Ballock RT, Heydemann A, Wakefield LM, Flanders KC, Roberts AB, Sporn MB. TGF-beta 1 prevents hypertrophy of epiphyseal chondrocytes: regulation of gene expression for cartilage matrix proteins and metalloproteases. Dev Biol 1993; 158 (02) 414-429
• 14 van Caam A, Madej W, Thijssen E. et al. Expression of TGFβ-family signalling components in ageing cartilage: age-related loss of TGFβ and BMP receptors. Osteoarthritis Cartilage 2016; 24 (07) 1235-1245
• 15 Liu Y, Hou R, Yin R, Yin W. Correlation of bone morphogenetic protein-2 levels in serum and synovial fluid with disease severity of knee osteoarthritis. Med Sci Monit 2015; 21: 363-370
• 16 Honsawek S, Chayanupatkul M, Tanavalee A. et al. Relationship of plasma and synovial fluid BMP-7 with disease severity in knee osteoarthritis patients: a pilot study. Int Orthop 2009; 33 (04) 1171-1175
• 17 Caron MM, Emans PJ, Cremers A. et al. Hypertrophic differentiation during chondrogenic differentiation of progenitor cells is stimulated by BMP-2 but suppressed by BMP-7. Osteoarthritis Cartilage 2013; 21 (04) 604-613
• 18 Bramlage CP, Häupl T, Kaps C. et al. Decrease in expression of bone morphogenetic proteins 4 and 5 in synovial tissue of patients with osteoarthritis and rheumatoid arthritis. Arthritis Res Ther 2006; 8 (03) R58
• 19 Kellgren JH, Lawrence JS. Radiological assessment of osteo-arthrosis. Ann Rheum Dis 1957; 16 (04) 494-502
• 20 Dominici M, Le Blanc K, Mueller I. et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006; 8 (04) 315-317
• 21 de Sousa EB, Dos Santos Junior GC, Aguiar RP. et al. Osteoarthritic Synovial Fluid Modulates Cell Phenotype and Metabolic Behavior In Vitro . Stem Cells Int 2019; 2019: 8169172
• 22 Miljkovic ND, Cooper GM, Marra KG. Chondrogenesis, bone morphogenetic protein-4 and mesenchymal stem cells. Osteoarthritis Cartilage 2008; 16 (10) 1121-1130
• 23 Carreira AC, Alves GG, Zambuzzi WF, Sogayar MC, Granjeiro JM. Bone Morphogenetic Proteins: structure, biological function and therapeutic applications. Arch Biochem Biophys 2014; 561: 64-73
• 24 Shioda M, Muneta T, Tsuji K. et al. TNFα promotes proliferation of human synovial MSCs while maintaining chondrogenic potential. PLoS One 2017; 12 (05) e0177771
• 25 Matsumura E, Tsuji K, Komori K, Koga H, Sekiya I, Muneta T. Pretreatment with IL-1β enhances proliferation and chondrogenic potential of synovium-derived mesenchymal stem cells. Cytotherapy 2017; 19 (02) 181-193
• 26 Zhang S, Muneta T, Morito T, Mochizuki T, Sekiya I. Autologous synovial fluid enhances migration of mesenchymal stem cells from synovium of osteoarthritis patients in tissue culture system. J Orthop Res 2008; 26 (10) 1413-1418
• 27 Kim YI, Ryu JS, Yeo JE. et al. Overexpression of TGF-β1 enhances chondrogenic differentiation and proliferation of human synovium-derived stem cells. Biochem Biophys Res Commun 2014; 450 (04) 1593-1599
• 28 Finnson KW, Chi Y, Bou-Gharios G, Leask A, Philip A. TGF-b signaling in cartilage homeostasis and osteoarthritis. Front Biosci (Schol Ed) 2012; 4: 251-268
• 29 Plaas A, Velasco J, Gorski DJ. et al. The relationship between fibrogenic TGFβ1 signaling in the joint and cartilage degradation in post-injury osteoarthritis. Osteoarthritis Cartilage 2011; 19 (09) 1081-1090