Additional file 2: Figure S2. of Combination of paclitaxel, bevacizumab and MEK162 in second line treatment in platinum-relapsing patient derived ovarian cancer xenografts

Body weight of animals treated with vehicle (CTR,-♦-), with Bevacizumab and MEK162 (BEV/MEK,-■-), paclitaxel and MEK162 (PTX/MEK,-▲-), paclitaxel and bevacizumab (PTX/BEV, -●-), or paclitaxel and bevacizumab and MEK162 (PTX/BEV/MEK,-x-). Data are expressed as mean±SD, each group consisted of 8-10 animals. A) MNHOC124 PDX model, B) MNHOC218 PDX model and C) MNHOC239 PDX model. (PDF 474 kb

<i>In vivo</i> effects of tivantinib and ZA single agent and in combination in a preventive schedule of bone metastases.

<p>(<b>A–B</b>) <i>In vivo</i> bioluminescence imaging (BLI) of 1833/TGL-i.c. injected athymic nude mice (4 week old) treated with tivantinib (120 mg/Kg; <i>n</i> = 7), ZA (100 mg/Kg; <i>n</i> = 5) and tivantinib+ZA (<i>n</i> = 5). (<b>A</b>) Ventral images from a representative nude mouse for each group 24 days after implant. Pseudocolor scale bars are consistent for all images of ventral views, in order to show relative changes at metastatic sites over time. (<b>B</b>) Average growth of bone metastasis in the hindlimbs of controls and treated mice: photon counts from the hindlimb (right and left) regions of tumor-bearing mice were quantified and displayed over time. The data are presented as mean ± SE. **, p<0.005; ***, p<0.0001. (<b>C</b>) The presence of tumor-induced osteolytic lesions were detected by weekly micro-CT scans. Representative 3D reconstruction of micro-CT images of the hindlimbs of controls and treated mice at day 24 after xenografting are reported. Similar results were obtained in the second experiment conducted independently. Red circles indicate osteolysis, the circle’s size was proportional to the extent of bone lesion. Numbers at the bottom of the images represent the number of mice. (<b>D</b>) Kaplan-Meier survival plot of the survival rate of 1833/TGL xenografted controls and treated-mice.</p

Effect of combination on <i>in vitro</i> cytotoxicity and cell migration.

<p>(<b>A</b>) In vitro cytotoxic activity of tivantinib alone (▪) or in combination with ZA (□) against 1833/TGL cells. Values represent the percentage of controls. (<b>B</b>) 1833/TGL cells were seeded at the concentration of 1×10⁵ cells/mL on a 12-well culture plate in six-well plates. On day 0, for each well, a wound was made in the center of the monolayer of confluent cells with a sterile plastic pipette tip and vehicle or drugs were added. The plate was placed under a motorized inverted microscope and wounds were photographed at 0 (<i>t</i>₀), 24, and 48 hours after wounding. Representative photographs of a filed of view from the different experimental conditions are shown. (<b>C</b>) Quantitative analysis: the level of cell migration was quantified as the percentage of wound closure at each time point after the wound scratch. Values represent averages ± SE of two independent experiments, each consisting of 2 replicates. Statistical analysis were performed at 24 and 48 hours after wounding.</p

Antitumor activity of tivantinib and ZA alone and in combination against subcutaneous breast cancer xenografts.

<p>A breast cancer xenograft model was established using 1833 cells (7.5×10⁶) implanted s.c. into the flanks of female athymic nude mice (6-week old). When tumor volume reached 70 mm³ animals were randomized into 4 groups: tivantinib (300 mg/Kg), ZA (100 mg/Kg), tivantinib+ZA-treated mice and controls treated with vehicle (PEG 400∶20% vit. E TPGS solution [60∶40]). Tivantinib and vehicle were administered p.o. in a volume of 10 mL/kg of body weight daily until the end of the experiment. ZA was administered i.p. every 2 days starting 13 days after implant till the end of the experiment. (<b>A</b>) Tumor growth inhibition. Tumor size was measured using Vernier caliper twice a week until the animals were sacrificed after 27 days of treatment. Tumor weight was calculated by the formula: Tumor weight (mg) = (length×width2)/2. (<b>B</b>) Relative body weight of mice bearing subcutaneous tumor xenografts and treated with vehicle or drugs. Body weights were measured twice weekly. Results are expressed as mean ± SD; <i>n</i> = 9–10.</p

Additional file 1: of The crosstalk between vascular MSCs and inflammatory mediators determines the pro-calcific remodelling of human atherosclerotic aneurysm

Figure S1. Angiogenic markers in AAA-MSCs before and after culture in matrigel. Significant decrease of CD146 mRNA can be observed after AAA-MSC differentiation in Matrigel; statistical differences were performed by two-way ANOVA with multiple comparisons among all experimental conditions. ****p < 0.0001. (TIF 1969 kb

Combination of the c-Met Inhibitor Tivantinib and Zoledronic Acid Prevents Tumor Bone Engraftment and Inhibits Progression of Established Bone Metastases in a Breast Xenograft Model

<div><p>Bone is the most common metastatic site for breast cancer. There is a significant need to understand the molecular mechanisms controlling the engraftment and growth of tumor cells in bone and to discover novel effective therapeutic strategies. The aim of this study was to assess the effects of tivantinib and Zoledronic Acid (ZA) in combination in a breast xenograft model of bone metastases. Cancer cells were intracardially implanted into immunodeficient mice and the effects of drugs alone or in combination on bone metastasis were evaluated by <i>in vivo</i> non-invasive optical and micro-CT imaging technologies. Drugs were administered either before (preventive regimen) or after (therapeutic regimen) bone metastases were detectable. In the preventive regimen, the combination of tivantinib plus ZA was much more effective than single agents in delaying bone metastatic tumor growth. When administered in the therapeutic schedule, the combination delayed metastatic progression and was effective in improving survival. These effects were not ascribed to a direct cytotoxic effect of the combined therapy on breast cancer cells <i>in vitro</i>. The results of this study provide the rationale for the design of new combinatorial strategies with tivantinib and ZA for the treatment of breast cancer bone metastases.</p></div

Scheme showing the experimental protocol used.

<p>Scheme showing the experimental protocol used.</p

<i>In vivo</i> effects of tivantinib and ZA alone and in combination in a therapeutic schedule of bone metastases.

<p>The bone metastatic 1833/TGL cells were injected into the left cardiac ventricle of 4-week old athymic nude mice (5×10⁵ cells/mouse). When bone metastases were detectable (11 days after cell implant), implanted animals were randomized into 4 groups of 10 mice each: controls treated with vehicle PEG 400∶20% vitamin E TPGS solution (60∶40) or treated groups administered with tivantinib alone (300 mg/Kg), ZA (100 mg/Kg) alone and combination of tivantinib plus ZA. Both vehicle and tivantinib were administered daily <i>per os</i> starting 13 days after implant until the end of the experiment. ZA was administered i.p. every 2 days starting 13 days after implant till the end of the experiment. (<b>A</b>) Representative optical scanning in the supine position for each group 27 days after tumor implant. The intensity of the BLI signal, measured as total photon flux, is shown as a pseudo-color scale bar. Numbers at the left bottom of the images represent the number of mice. (<b>B</b>) At the indicated times after xenografting, the BLI signal was captured and the growth kinetics of right and left hindlimb metastasis for each group were expressed in the graph as mean value of total photon counts from the hindlimbs of animals. Data are presented as mean value ± SE; <b>*</b>, p<0.05 tivantinib+ZA vs tivantinib. (<b>C</b>) Representative volume-rendered micro-CT images of osteolytic bone metastases obtained 27 days after xenografting are shown. Red circles indicate osteolysis and the numbers at the bottom of the images represent the number of mice. (D) Kaplan–Meyer curves of vehicle and treated groups.</p

Dynamic compliance (C_dyn) of surfactant-depleted rabbits managed with non-invasive ventilation.

<p>To the left of the dashed line, the baseline and post-surfactant depletion C_dyn values of all the animals featured in the study are given. To the right of the dashed line, the C_dyn values of the different groups after 180 minutes of management with NIV are shown. Surfactant-treated groups are represented by the box-plots with a grey filling, whereas the groups merely treated with NIV are represented by the white box-plots. The small squares within each box-plot indicate the mean of the group. The whiskers indicate the maximum and minimum values observed for each group. ^# <i>P</i> < 0.01 vs. SNIPPV+SF group. NCPAP: nasal continuous positive airway pressure, SF: surfactant, NIPPV: nasal (non-synchronized) intermittent positive pressure ventilation, and SNIPPV: synchronized NIPPV.</p

Histological scores of surfactant-depleted adult rabbits.

<p>Histological scores of surfactant-depleted adult rabbits.</p