Journal: Nature Communications

Article Title: Regulation of heterotopic ossification by monocytes in a mouse model of aberrant wound healing

doi: 10.1038/s41467-019-14172-4

Figure Lengend Snippet: a Schematic of experimental set-up. b Flow cytometry analysis of injury site 1 week after burn/tenotomy in mice treated with clodronate or PBS for monocytes (CD11b + Ly6G − ); neutrophils (CD11b + Ly6G + ); classical monocytes (CD11b + Ly6G − Ly6C hi ), alternatively activated monocytes (CD11b + Ly6G − Ly6C lo ) and macrophages (F4/80 + ). Circulating monocytes: n = 4/group, df = 8, t = −0.213, p = 0.000. Injury site: n = 4/group, df = 8. Neutrophils: t = −0.213,p = 0.837; Monocytes: t = 3.490, p = 0.008; Ly6C − : t = 3.193, p = 0.013; Ly6C low : t = 4.139, p = 0.003; Ly6C hi : t = −0.979, p = 0.356; F4/80 macrophages: t = 3.552, p = 0.007. c IVIS imaging of MPO activity 1 week after injury in mice treated with clodronate or control ( n = 4 mice per treatment). Total bioluminescent signal at the injury site using standardized region of interest (ROIs) was calculated and presented as total flux in photons per second per ROI. df = 3.182, t = 1.995, p = 0.135. d Left: representative images of ankle edema present in each treatment group. Right: quantification of ankle size. n (PBS) = 5, n (Clod) = 4, df = 7, t = 11.350, p = 0.000. e Representative Safranin O staining of tendon injury site 3 weeks after burn/tenotomy in clodronate and PBS treated mice. n = 3/group. f MicroCT analysis of tenotomy site 9 weeks after burn/tenotomy in clodronate and PBS control treated mice. Left: representative 3D reconstruction. Right: quantification of unthresholded total HO, floating HO (HO not associated with tibia or calcaneus) and proximal HO (HO proximal to the calcaneus). n (PBS) = 6, n (Clod) = 5. Total HO: t = 3.302, df = 5.312, p = 0.020; Floating HO: t = 1.867, df = 5.002, p = 0.121; Proximal HO: t = 1.313, df = 9, p = 0.222. All analyses assess for homoscedasticity and difference in means via Levene’s F-test and two-tailed Student’s t-test, respectively. * p < 0.05. Source data are provided as a Source Data file.

Article Snippet: Mouse hind limbs were harvested and imaged 9-weeks post-injury (Bruker MicroCT, 35 μm resolution and 357 μA/70 kV/beam).

Techniques: Flow Cytometry, Imaging, Activity Assay, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Regulation of heterotopic ossification by monocytes in a mouse model of aberrant wound healing

doi: 10.1038/s41467-019-14172-4

Figure Lengend Snippet: a Representative images of IVIS analysis for MPO of C57BL/6 J and LysMCre- Tgfb1 fl/fl mice bred on a C57BL/6 J background at 1 week after burn/tenotomy. Right: quantification of total bioluminescence/region of interest. n = 5/group. Injury site: t = 1.898, df = 8, p = 0.094; dorsum: t = 6.385, df = 4.007, p = 0.003. b Quantification of neutrophils and monocyte subpopulations based on Ly6C using flow cytometry of injury site 1 week after burn/tenotomy in C57BL/6 J and LysMCre- Tgfb1 fl/fl mice bred on a C57BL/6 J background. n = 4 mice/group. n = 3/group. Neutrophils: t = −1.125, fd = 4, p = 0.324; Ly6C - : t = 1.685, df = 4, p = 0.167; Ly6C lo : t = 0.315, df = 4, p = 0.768; Ly6C hi : t = −1.272, df = 4, p = 0.272. c MicroCT analysis of tenotomy site 9 weeks after burn/tenotomy in C57BL/6 J and LysMCre- Tgfb1 fl/fl mice. Left: Representative 3D reconstruction. Right: Quantification of total HO, floating HO (not associated with tibia or calcaneus) and proximal HO (all HO proximal to calcaneus). n = 5/group. Total HO: t = 2.290, df = 0, p = 0.051; Floating HO: t = 4.591, df = 4, p = 0.008; Proximal HO: t = 2.578, df = 8, p = 0.033. d Levels of TGFβ1 in pg/ug total protein from homogenates from the extremity injury and plasma from LysMCre- Tgfb1 fl/fl , LysMCre- Tgfb1 fl/wt , or wild type mice 3 days after burn/tenotomy n = 3 mice in each genotype. n = 3/group, df across groups = 2. F statistics: homogenate TGFB1: 0.588, plasma TGFB1: 1.008. All pairwise comparisons were analyzed for homoscedasticity and difference in means via Levene’s F-test and two-tailed Student’s t-test, respectively. Homoscedastic and heteroscedastic multi-group analyses were performed via ANOVA + post-hoc Dunnett’s test and Welch’s comparison of means + post-hoc Dunnett’s T3 test, respectively. * p < 0.05, ** p < 0.01. Source data are provided as a Source Data file.

Article Snippet: Mouse hind limbs were harvested and imaged 9-weeks post-injury (Bruker MicroCT, 35 μm resolution and 357 μA/70 kV/beam).

Techniques: Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Regulation of heterotopic ossification by monocytes in a mouse model of aberrant wound healing

doi: 10.1038/s41467-019-14172-4

Figure Lengend Snippet: a Representative Safranin O stain of tendon injury site 3 weeks after burn/tenotomy in p7N3 (CD47 agonist) treated and PBS control mice. n = 3/group. b MicroCT analysis of tenotomy site 9 weeks after burn/tenotomy in PBS and p7N3 (CD47 agonist) treated mice. Left: Representative 3D reconstruction. Right: Quantification of total HO, floating HO and proximal HO. n = 7/group. Total HO: t = 3.415, df = 7.840, p = 0.009; Floating HO: t = 2.201, df = 12, p = 0.048; Proximal HO: t = 2.686, df = 8.549, p = 0.026. c Levels of TGF-β1, TGFβ2, and TGFβ3 in pg/ug total protein and represented as median with interquartile range from Top: homogenates from the extremity injury (TGF-β1: t = −0.635, df = 4, p = 0.560; TGF-β2: t = −0.643, df = 4, p = 0.555; TGF-β3: t = −1.272, df = 2.186, p = 0.322) and Bottom: plasma from PBS and p7N3 (CD47) peptide treated mice 3 days after burn/tenotomy (TGF-β1: t = 1.544, df = 2.037, p = 0.260; TGF-β2: t = 2.747, df = 4, p = 0.052; TGF-β3: t = −1.492, df = 4, p = 0.210). n = 3 mice per treatment group. d qPCR analysis of M1 ( iNos ) and M2 ( Arg1 and Mrc1 ) macrophage markers and Tgfb1 in macrophages isolated from the extremity injury site of naive (day 0), burn/tenotomy day 3, burn/tenotomy day 3 treated with PBS, and burn/tenotomy day 3 treated with p7N3 (CD47) peptide. Day 0 vs. Day 3— iNOS : t = 2.020, df = 2, p = 0.181; Arg1 : t = −6.084, df = 3, p = 0.009; Mrc1 : t = 0.703, df = 4, p = 0.521; Tgfb1 : t = 0.253, df = 4, p = 0.812. PBS vs. CD47 – iNOS : t = −0.834, df = 2.043, p = 0.491; Arg1 : t = 0.895, df = 4, p = 0.421; Mrc1 : t = 1.176, df = 4, p = 0.305; Tgfb1 : t = 1.186, df = 4, p = 0.301. e Representative images of phagocytosis assay using macrophages isolated from the extremity injury at day 3 after burn/tenotomy in mice treated with PBS or p7N3 (CD47). PBS n = 3, CD47 n = 4 approximately 25 cells/mouse. f Measurement of cellular circularity Circularity: t = 6.119, df = 55.537, p = 0.000 and quantification of mean fluorescent intensity phagocytosed by each macrophage. MFI: t = −0.357, df = 111, p = 0.722. Source data are provided as a Source Data file.

Article Snippet: Mouse hind limbs were harvested and imaged 9-weeks post-injury (Bruker MicroCT, 35 μm resolution and 357 μA/70 kV/beam).

Techniques: Staining, Isolation, Phagocytosis Assay

Journal: bioRxiv

Article Title: Full-field strain distribution in hierarchical electrospun nanofibrous Poly-L(lactic) acid and Collagen based scaffolds for tendon and ligament tissue regeneration: a multiscale study

doi: 10.1101/2023.06.01.543145

Figure Lengend Snippet: Workflow of the microCT morphometric analysis. A) EHS transversal section and rendering highlighting total area (contoured in yellow) and internal area (contoured in green); B) EHS transversal section and volume rendering highlighting objects (in red) running in bundles (scale bar = 1 mm).

Article Snippet: Each sample was immersed in PBS for 2 min (SB and EHS gauge length = 10 mm) before it was clamped in the microCT loading device (maximum actuator displacement = 5.5 mm; displacement rate = 0.001 mm s -1 ) (MTS in situ tester for Skyscan 1172, Bruker, Belgium).

Techniques: