Inflammatory stimuli differentially modulate the transcription of paracrine signaling molecules of equine bone marrow multipotent mesenchymal stromal.

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Presentation transcript:

Inflammatory stimuli differentially modulate the transcription of paracrine signaling molecules of equine bone marrow multipotent mesenchymal stromal cells R. Vézina Audette, A. Lavoie-Lamoureux, J.-P. Lavoie, S. Laverty Osteoarthritis and Cartilage Volume 21, Issue 8, Pages 1116-1124 (August 2013) DOI: 10.1016/j.joca.2013.05.004 Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 eBM-MSCs' expression of paracrine pro-inflammatory and modulatory signaling molecules implicated in the pathology of OA is influenced by stimulation with LPS or IL-1β in vitro. LPS significantly increased the transcription of (A) TNF-α at 1 h (23 fold (9.6–56.4), P < 0.0001, n = 3) and 3 h (seven fold (2.3–19.6), P = 0.001, n = 2), (C) IL-8 at all timepoints (1 h: nine fold (3.6–22.9), P < 0.0001, n = 3, 3 h: nine fold (3.2–27.3), P = 0.0003, n = 2, 6 h: five fold (1.9–12.3), P = 0.002, n = 3, 12 h: 13 fold (5.0–32.2), P < 0.0001, n = 3, 24 h: six fold (2.2–14.5), P = 0.0008, n = 3) and (D) IL-6 at 3 h (510 fold (19.1–13638.4), P = 0.0007, n = 2) compared to unstimulated eBM-MSCs but (B) IL-1β expression was not affected by this stimulation. Conversely, IL-1β stimulation significantly increased the transcription of (B) IL-1β at 1 h (10 fold (2.9–31.6), P = 0.0007, n = 3) and 12 h (11 fold (3.3–35.1), P = 0.0005, n = 3) and (C) IL-8 at 1 h (five fold (1.9–11.9), P = 0.002, n = 3), 3 h (seven fold (2.3–19.8), P = 0.001, n = 2), 12 h (nine fold (3.3–21.5), P = 0.0001, n = 3) and 24 h (10 fold (3.9–25.3), P < 0.0001, n = 3) but (A) TNF-α and (D) IL-6 expression was unchanged by IL-1β stimulation compared to unstimulated eBM-MSCs. Open circles represent LPS-treated eBM-MSCs, solid circles represent IL-1B-treated eBM-MSCs and * represent statistically significant fold increases (P ≤ 0.05). Osteoarthritis and Cartilage 2013 21, 1116-1124DOI: (10.1016/j.joca.2013.05.004) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 eBM-MSCs' expression of the paracrine anabolic signaling molecules implicated in the pathology of OA is unaffected by stimulation with LPS or IL-1β. (A) VEGF, (B) TGF-β1 and (C) IGF-1 gene expression is not significantly different in cultured eBM-MSCs stimulated with LPS or IL-1β compared to unstimulated eBM-MSCs. Open circles represent LPS-treated eBM-MSCs, solid circles represent IL-1B-treated eBM-MSCs and * represent statistically significant fold increases (P ≤ 0.05). Osteoarthritis and Cartilage 2013 21, 1116-1124DOI: (10.1016/j.joca.2013.05.004) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 eBM-MSCs' expression of paracrine pro-inflammatory and modulatory signaling molecules implicated in the pathology of OA is influenced by stimulation with NSF or OASF. (A) TNF-α, (C) IL-8 and (D) IL-6 gene expression at all timepoints was not significantly different in cultured eBM-MSCs stimulated with SFs from either normal or OA joints. However, the transcription of (B) IL-1β at 24 h was significantly decreased upon stimulation with both NSF and OASF (NSF: 0.10 fold decrease (0.031–0.33), P = 0.0006, n = 3; OASF; 0.07 fold decrease (0.023–0.24), P = 0.0002, n = 3) compared to unstimulated eBM-MSCs. Open triangles represent NSF-treated eBM-MSCs, solid triangles represent OASF-treated eBM-MSCs and * represent statistically significant fold decreases (P ≤ 0.05). Osteoarthritis and Cartilage 2013 21, 1116-1124DOI: (10.1016/j.joca.2013.05.004) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 eBM-MSCs' expression of the paracrine anabolic signaling molecule VEGF is significantly up-regulated on exposure to OASF for 1 h. (A) VEGF levels are significantly increased upon stimulation with OASF at 1 h (three fold (1.49–4.97), P = 0.002, n = 3) but not longer stimulations whereas (B) TGF-β1 and (C) IGF-1 levels are not significantly different in cultured eBM-MSCs stimulated with either NSF or OASF compared to unstimulated controls at any timepoint. Open triangles represent NSF-treated eBM-MSCs, solid triangles represent OASF-treated eBM-MSCs and * represent statistically significant fold increases (P ≤ 0.05). Osteoarthritis and Cartilage 2013 21, 1116-1124DOI: (10.1016/j.joca.2013.05.004) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions