Figure 4.

Effect of thrombospondin-1 (THBS1) on inflammatory cells. (A) Levels of pro-inflammatory proteins (IL-6, IL-8, MCP-1, MIP1, and RANTES) were quantified in the supernatants of synoviocytes cultured with or without adipose stem cell (ASC)-siCT or ASC-siTHBS1. Results are expressed as the mean concentration of cytokine ± SEM (n = 3 biological replicates). (B) Pro-inflammatory cytokine expressions (IL-6, TNFα, and IL-8) was quantified in osteoarthritic chondrocytes after coculture with ASC-siCT or ASC-siTHBS1 (n = 4 biological replicates). Results are expressed as fold change of gene expression compared to untreated chondrocytes or chondrocyte monocultures and represented as mean ± SEM. (C) Role of THBS1 on T cell proliferation was quantified using murine splenocytes activated with ConA in presence of different concentrations of recombinant THBS1 (rTHBS1) or (D) activated murine splenocytes in presence of ASC-siCT or ASC-siTHBS1 at different ratios. Results are expressed as the percentage of ConA-induced proliferation of splenocytes which was assigned the value of 100% and represented as mean ± SEM (n = 6 independent biological replicates). (E) Immunosuppressive molecule expression (TSG6, IL-6, COX2, and IDO1) was quantified in ASC-siCT and ASC-siTHBS1. Results are expressed as relative gene expression (2^−ΔCT) and represented as mean ± SEM (n = 8 independent biological replicates). Statistics used Mann–Whitney test (A,B,E) or Kruskal–Wallis (C,D): *p < 0.05, **p < 0.01, ***p < 0.001 compared to control.