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Results: 6

1.
Figure 6

Figure 6. Mechanical results of the healing MCL after IL-1Ra treatment.. From: Interleukin Expression after Injury and the Effects of Interleukin-1 Receptor Antagonist.

Graphs show failure force (a), stiffness (b), and failure stress (c) of the MCL 11 days post-injury after PBS or IL-1Ra treatment. No significance was observed in any tested parameter (p>.05). Results are expressed as mean ± S.E.M.

Connie S. Chamberlain, et al. PLoS One. 2013;8(8):e71631.
2.
Figure 3

Figure 3. Representative micrographs of the healing MCL after PBS or IL-1Ra treatment.. From: Interleukin Expression after Injury and the Effects of Interleukin-1 Receptor Antagonist.

Micrographs demonstrate H&E staining (a–b), and immunohistochemistry of M2 macrophages (c–f), and proliferating cells (g–h) after PBS (left column) or IL-1Ra (right column) treatment. H&E staining of the day 11 MCL indicating larger granulation tissue size (a–b). M2 macrophages 5 days- (c–d) and 11 days post-injury (e–f) increase in number after IL-1Ra treatment. In contrast, proliferating cells decrease on day 5 (g–h) after IL-1Ra administration.

Connie S. Chamberlain, et al. PLoS One. 2013;8(8):e71631.
3.
Figure 1

Figure 1. Microarray results identifying specific interleukins modulated at days 3 and/or 7 post-injury.. From: Interleukin Expression after Injury and the Effects of Interleukin-1 Receptor Antagonist.

A total of 12 interleukins were significantly modulated (2 fold greater than normal and q-value <.05) compared to the intact control. Of those 12, 6 were up-regulated at day 3 only (il6, il7, il17r, 1l18, il1b, and il1rn), 1 up-regulated at day 7 only (il13ra1), 1 down-regulated at day 7 only (il11ra1), 2 up-regulated at both day 3 and 7 (il1rl1, il11), and 1 down-regulated at both day 3 and 7 (il16). Interleukins regulated during healing were both pro- and anti-inflammatory (a). Members of the il1 family were significantly regulated with healing (b). Microarray data are compared to the normal intact ligament and expressed as fold change ± S.D. * indicate fold change ≥2 from the intact control.

Connie S. Chamberlain, et al. PLoS One. 2013;8(8):e71631.
4.
Figure 5

Figure 5. Luminex results of cytokines regulated by IL-1Ra.. From: Interleukin Expression after Injury and the Effects of Interleukin-1 Receptor Antagonist.

Graphs show the effects of PBS and IL-1Ra on MCL-derived IL-1α (a), IL-1β (b), IL-12 (c), IL-2 (d), IFN-γ (e) IL-10 (f), and IL-6 (g) production. Blockade of the IL-1 activation resulted in an accumulation of IL-1α (a) and IL-1β (b). By day 11, inhibition of the IL-1R resulted in a down-regulation of IL-1α (a) and tended to decrease IL-1β (b; p = .056). IL-1Ra significantly decreased the pro-inflammatory cytokines, IL-12 (c), IL-2 (d), and IFN-γ (e) on day 5. IL-12 levels remained significantly different after IL-1Ra on day 11. The anti-inflammatory cytokine, IL-10 was up-regulated at day 5 after IL-1 inhibition (f). IL-6 was not significantly influenced by treatment (f). Beneath graph, p value indicates ANOVA results. a,b,c indicates within a graph, bars without a common superscript letter differ (results of Fisher’s LSD post-hoc pairwise analysis, p<.05). *indicates p value.056. Values are expressed as normalized cytokine concentration (ng cytokine/mg tissue weight) ± S.E.M.

Connie S. Chamberlain, et al. PLoS One. 2013;8(8):e71631.
5.
Figure 2

Figure 2. Immunohistochemistry results of selected cellular factors regulated by IL-1Ra.. From: Interleukin Expression after Injury and the Effects of Interleukin-1 Receptor Antagonist.

Graph of the percent healing region (a), granulation tissue localized M1 macrophages (b), granulation tissue-localized M2 macrophages (c), M2 macrophages outside the granulation tissue (d), and granulation tissue-localized proliferating cells (d), at 5 and 11 days post-injury after PBS or IL-1Ra treatment. On day 11, IL-1Ra significantly increased the percent healing region (a). No differences within the MCL were observed at any other points. Although no differences were observed with M1 macrophages (b), IL-1Ra treatment increased M2 macrophage numbers at day 5 and/or day 11 within (c) and outside of the granulation tissue (d). Proliferating cells were significantly reduced at day 5 (e). a,b,c indicates within a graph, bars without a common superscript letter differ (results of Fisher’s LSD post-hoc pairwise analysis, p<.05). Values are expressed as mean cell numbers ± S.E.M.

Connie S. Chamberlain, et al. PLoS One. 2013;8(8):e71631.
6.
Figure 4

Figure 4. Immunohistochemistry results of cellular and ECM factors regulated by IL-1Ra.. From: Interleukin Expression after Injury and the Effects of Interleukin-1 Receptor Antagonist.

Immunohistochemistry results showing the effects of PBS and IL-1Ra on granulation tissue localized endothelial cells (a), granulation tissue-localized blood vessel lumen (b), total myofibroblasts (c), type I procollagen (d), and type III collagen (e) at 5 and 11 days post-injury. IL-1Ra significantly increased the number of endothelial cells and blood vessel lumen on day 5 (a–b), but not day 11. In contrast, IL-1Ra significantly decreased the number of myofibroblasts at day 5 (c). No differences within the MCL were observed on day 11 (c). Regardless of day, no treatment differences were observed with type I procollagen (d) and type III collagen (e). ). a,b indicates within a graph, bars without a common superscript letter differ (results of Fisher’s LSD post-hoc pairwise analysis, p<.05). Values are expressed as mean density ± S.E.M. Representative micrographs of endothelial cells/blood vessel lumen (f–g) and myofibroblasts (h–i) 5 days post-injury after PBS (left column) and IL-1Ra (right column) treatment.

Connie S. Chamberlain, et al. PLoS One. 2013;8(8):e71631.

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