Additional file 5: of Surgical management of chronic lateral ankle instability: a meta-analysis

Characteristics of included studies 4 (PDF 93 kb

Additional file 7: of Surgical management of chronic lateral ankle instability: a meta-analysis

Characteristics of included studies 6 (PDF 91 kb

Additional file 1: of Surgical management of chronic lateral ankle instability: a meta-analysis

Search strategies (DOCX 14 kb

Additional file 2: of Surgical management of chronic lateral ankle instability: a meta-analysis

Characteristics of included studies 1 (PDF 88 kb

Additional file 9: of Surgical management of chronic lateral ankle instability: a meta-analysis

Characteristics of excluded studies (PDF 90 kb

Cytokine-stimulated superoxide flashes in chondrocytes <i>in situ</i>.

<p><b>A</b><b>B</b>, Time-lapse views of a typical superoxide flash <i>in situ</i> in the red boxed region of 2.5 µm×5 µm. Time course plot of the superoxide flash is shown at the bottom. Note the swelling of the mitochondrion during the flash. <b>C</b>, Activity and properties of chondrocyte superoxide flashes i<i>n vitro</i> (510 events as in Figure. 2) and <i>in situ</i> (30 events). Data are reported as the mean ± SEM values. *, <i>p</i><0.05; **, <i>p</i><0.01. <b>D</b>, Time course of the superoxide flash response to cytokine stimulation. Data are reported as the mean ± SEM values. n = 21–29 imaging planes. *, <i>p</i><0.05; **, <i>p</i><0.01 versus the respective control.</p

Imaging chondrocytes and mitochondria <i>in situ</i>.

<p><b>A</b>, Confocal or two-photon-excitation imaging of chondrocyte mitochondria in femoral head cartilage <i>in situ</i>. Insert shows an enlarged view of the femoral head. <b>B</b>, Mitochondrial distribution in intact cartilage. The femoral head was sliced lengthwise along the midcoronal panel, labeled with TMRM and imaged by a confocal microscope. Arrows mark the cartilage surface and arrowheads mark the growth plate. <b>C</b>, Two-photon excitation images of TMRM-stained mitochondria (excitation at 850 nm, red) in chondrocytes at different depths from the cartilage surface. Layer thickness: 0.77 µm, XYZ of 3D: 100*100*100 µm³ (Z axis shows depth). <b>D</b>, Profiles of averaged mitochondrial (Mito, green) and chondrocyte (Cell, blue) cross-section areas and their ratios (Mito/Cell, red) as a function of distance from the surface. Right panels shows representative images from superficial, middle and deep cartilage zones.</p

Mitochondrial superoxide flashes in cultured articular chondrocytes.

<p><b>A</b>, Colocalization of the superoxide biosensor – mitochondrial circularly permuted yellow fluorescent protein (mt-cpYFP) and tetramethylrhodamine methyl ester (TMRM) in chondrocyte mitochondria revealed by tri-wavelength excitation imaging. Chondrocytes were isolated from mt-cpYFP transgenic mice and cultured as described in the Methods. Only chondrocytes from the first passage were used for experiments. <b>B</b>, Mitochondrial superoxide flashes in articular chondrocytes. Enlarged (top panel) and pseudo-color contrast-enhanced (middle panel, scale bar shown to the bottom left) snapshots of a representative mitochondrial superoxide flash in chondrocyte are shown to the right. The lower panel shows the time course of the corresponding mt-cpYFP fluorescence intensity at 488 nm excitation. <b>C</b>, Histogram analysis of amplitude (ΔF/F₀) and full duration at half maximum (FDHM) of chondrocyte superoxide flashes. <b>D</b>, Kinetics of superoxide flashes. Note the characteristic lack of fluorescence change at 405 nm excitation. Superoxide flash coincided with transient mitochondrial depolarization (ΔΨ_m, TMRM at 543 nm excitation).</p

Enhanced superoxide flash activity and mitochondrial fragmentation during IL-1β or TNF-α challenge in cultured chondrocytes.

<p>A, Representative examples showing superoxide flash activities in basal conditions and after IL-1β or TNF-α challenges in different chondrocytes. Data are presented as an overlay of the XY view of a chondrocyte (bottom) and surface plot of all superoxide flashes combined from 100 consecutive frames obtained at 1 frame/s. The lower panel shows temporal diaries of superoxide flash incidence in nine representative cells belonging to three groups respectively, with the uppermost diary corresponding to the image shown. B, Superoxide flash frequency without or during IL-1β (10 ng/ml) or TNF-α (10 ng/ml) challenge. Data are shown as the mean ± SEM values. n = 179–576 events from 42–49 cells for each group. **, <i>p</i><0.01 versus the corresponding control group. C, Characteristics of spontaneous (control) and cytokine-stimulated (IL-1β or TNF-α) chondrocytes superoxide flashes. ΔF/F₀: amplitude. FDHM: full duration at half maximum. FAHM: full area at half maximum. Data represent the mean ± SEM values. n = 42–49 cells for each group. *, <i>p</i><0.05; **, <i>p</i><0.01 versus the corresponding control group. D, Representative chondrocyte mitochondrial morphology before (control) and 60 min after IL-1β or TNF-α treatment.</p