BMP-4, TGF-β e Smad3 como moduladores da viabilidade das células do líquido sinovial^[*]

Resumo

Objetivo Nosso objetivo foi avaliar a modulação das células do líquido sinovial (SFCs, na sigla em inglês) de pacientes com e sem osteoartrite (OA) por proteína morfogenética óssea 4 (BMP-4), Smad3 e transformador do fator de crescimento β (TGF-β).

Métodos O do líquido sinovial foi coletado de pacientes submetidos a artroscopia ou artroplastia do joelho, e centrifugados para isolar as células do liquido sinovial. As células foram cultivadas por 21 dias e caracterizadas como células-tronco mesenquimais (MSCs, na sigla em inglês) de acordo com os critérios da International Society of Cell Therapy. Em seguida, realizamos um ensaio de brometo de 3-4,5-dimetil-tiazol-2-il-2,5-difeniltetrazólio (MTT) depois de expor células com e sem OA para TGF-β, inibidores de via Smad3 e BMP-4 e para diferentes concentrações de BMP-4.

Resultados A exposição aos inibidores TGF-β, Smad3 e BMP-4 modifica a atividade mitocondrial das SFCs. A atividade das SFCs é modificada por influências sobre o aumento das concentrações de BMP-4, mas não há diferença na atividade celular entre pacientes com e sem OA.

Conclusão TGF-β, Smad3 e BMP-4 modulam a atividade das SFCs de pacientes com e sem OA do joelho.

Palavras-chave

células-tronco mesenquimais - osteoartrite - células do líquido sinovial - caminho TGF-β

• 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