Antiangiogenic effects of decorin restored by unfractionated, low molecular weight, and nonanticoagulant heparins by Amy K. L. Chui, Tilini N. Gunatillake,

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Presentation transcript:

Antiangiogenic effects of decorin restored by unfractionated, low molecular weight, and nonanticoagulant heparins by Amy K. L. Chui, Tilini N. Gunatillake, Vera Ignjatovic, Paul T. Monagle, Padma Murthi, Shaun P. Brennecke, John M. Whitelock, and Joanne M. Said BloodAdv Volume 1(16):1243-1253 July 11, 2017 © 2017 by The American Society of Hematology

Amy K. L. Chui et al. Blood Adv 2017;1:1243-1253 © 2017 by The American Society of Hematology

Treatment with all 4 concentrations of UFH and LMWH decreased thrombin generation levels of control and DCN-reduced TIME cells, with the exception of NAC. The ETP of the TIME cells after reduction of DCN and incubated with heparins was determined using the CAT system. Treatment with all 4 concentrations of UFH and LMWH decreased thrombin generation levels of control and DCN-reduced TIME cells, with the exception of NAC. The ETP of the TIME cells after reduction of DCN and incubated with heparins was determined using the CAT system. Representative images from the CAT output are also shown for each heparin used, where the y-axis represents the ETP in nanomolars and the x-axis shows the time in minutes. (A) Treatment without heparin and treatment with all 4 concentrations of UFH (A), LMWH (B), and NAC (C) in a dose-dependent manner. *P < .01, ****P < .00001 (both significant); n = 9; 1-way ANOVA. The y-axis represents the ETP (nanomolars per minute). Amy K. L. Chui et al. Blood Adv 2017;1:1243-1253 © 2017 by The American Society of Hematology

Treatment with all 4 concentrations of UFH restored the network-formation abilities of DCN-reduced TIME cells. Treatment with all 4 concentrations of UFH restored the network-formation abilities of DCN-reduced TIME cells. The ability of TIME cells to form networks after DCN siRNA or Mock/NC was determined using the μ-Slide Angiogenesis system by IBIDI. (A) Representative image of the networks stained with calcein-AM is shown for each concentration. Images were acquired using the Olympus BX53 fluorescence microscope and represent 10× magnification to obtain the whole field. (B) The network-formation potential of TIME cells after treatment with 4 concentrations of UFH calculated by the Wimasis software. *P < .05, **P < .001, ***P < .0001, ****P < .00001 (all significant); n = 9; 1-way ANOVA. The y-axis represents the number of branch points. Amy K. L. Chui et al. Blood Adv 2017;1:1243-1253 © 2017 by The American Society of Hematology

Treatment with all 4 concentrations of LMWH restored the network-formation abilities of DCN-reduced TIME cells. Treatment with all 4 concentrations of LMWH restored the network-formation abilities of DCN-reduced TIME cells. (A) Representative image of the networks stained with calcein-AM is shown for each concentration. Images were acquired using the Olympus BX53 fluorescence microscope and represent 10× magnification to obtain the whole field. (B) The network-formation potential of TIME cells after treatment with LMWH calculated by the Wimasis software. **P < .001, ***P < .0001, ****P < .00001 (all significant); n = 9; 1-way ANOVA. The y-axis represents the number of branch points. Amy K. L. Chui et al. Blood Adv 2017;1:1243-1253 © 2017 by The American Society of Hematology

Treatment with all 4 concentrations of NAC restored the network-formation abilities of DCN-reduced TIME cells. Treatment with all 4 concentrations of NAC restored the network-formation abilities of DCN-reduced TIME cells. (A) Representative image of the networks stained with calcein-AM is shown for each concentration. Images were acquired using the Olympus BX53 fluorescence microscope and represent 10× magnification to obtain the whole field. (B) Network formation of TIME cells after treatment with NAC calculated by the Wimasis software. *P < .05, **P < .001 (both significant); n = 9; 1-way ANOVA. The y-axis represents the number of branch points. Amy K. L. Chui et al. Blood Adv 2017;1:1243-1253 © 2017 by The American Society of Hematology

Treatment with all 4 concentrations of UFH or LMWH, but not NAC, reduced cell growth of DCN-reduced TIME cells. Treatment with all 4 concentrations of UFH or LMWH, but not NAC, reduced cell growth of DCN-reduced TIME cells. The effect on TIME-cell proliferation after 48 hours of incubation with DCN siRNA or Mock/NC incubated in 4 concentrations of different heparins was determined using the xCELLigence system. Representative images from the xCELLigence output are also shown for each heparin used, where the y-axis represents the cell index and the x-axis shows the time in minutes. (A) A representative graph showing the cell index of TIME cells treated with DCN siRNA or Mock/NC incubated with 0.15, 0.35, 0.5, and 1.0 IU of UFH over 48 hours in culture. ****P < .00001 (significant); n = 9; 1-way ANOVA. The y-axis represents the cell index at 48 hours. (B) The results after incubation with 0.15, 0.5, 0.7, and 1.2 IU of LMWH. ***P < .0001, ****P < .00001 (both significant); n = 9; 1-way ANOVA. The y-axis represents the cell index at 48 hours. (C) Treatment with 0.15, 0.4, 0.6, and 1.1 IU of NAC and cell growth in Mock/NC and DCN-reduced cells. *P < .05, **P < .001, ***P < .0001 (all significant); n = 9; 1-way ANOVA. The y-axis represents the cell index at 48 hours. Amy K. L. Chui et al. Blood Adv 2017;1:1243-1253 © 2017 by The American Society of Hematology