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Angiogenic properties of sustained release platelet-rich plasma: Characterization in-vitro and in the ischemic hind limb of the mouse  Shyamal Chandra.

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Presentation on theme: "Angiogenic properties of sustained release platelet-rich plasma: Characterization in-vitro and in the ischemic hind limb of the mouse  Shyamal Chandra."— Presentation transcript:

1 Angiogenic properties of sustained release platelet-rich plasma: Characterization in-vitro and in the ischemic hind limb of the mouse  Shyamal Chandra Bir, MBBS, PhD, Jiro Esaki, MD, Akira Marui, MD, PhD, Kenichi Yamahara, MD, PhD, Hideki Tsubota, MD, Tadashi Ikeda, MD, PhD, Ryuzo Sakata, MD  Journal of Vascular Surgery  Volume 50, Issue 4, Pages e2 (October 2009) DOI: /j.jvs Copyright © 2009 Society for Vascular Surgery Terms and Conditions

2 Fig 1 Platelet-rich plasma (PRP) contains a large number of platelets in a small volume of plasma compared with platelet-poor plasma (PPP). (A) Shows the concentration of Platelets in the PPP and that in the PRP. *P < .05 vs group PPP. (B-F) Represents the concentration of different growth factors in platelet-poor plasma (PPP) and that in the platelet-rich plasma (PRP); stromal cell derived factor (SDF-1α, n = 5; B), Platelet derived growth factor (PDGF-BB, n = 5; C), Vascular endothelial growth factor (VEGF, n = 5; D), basic fibroblast growth factor (bFGF, n = 6; E), and insulin like growth factor (IGF-1, n = 5; F). *P < .05 vs group PPP. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Platelet-rich plasma (PRP) induces endothelial cell proliferation and capillary tube formation. (A) Dose response of PRP on endothelial cell proliferation; n = 3 in each group. Groups C = control, 1 μL = 1 μL of PRP, 10 μL = 101 μL of PRP, 100 μL = 100 μL of PRP, n = 3 in each group. (B) Time course of endothelial cell proliferation resulted by control (C) and PRP (10 μL). (C) Capillary tube area formed by control (C) and PRP, n = 3 in each group. (D-E) Photomicrographs of capillary tube by control (0.5% FBS) and PRP. Scale bars indicate 100 μm. *P < .05 vs group C, †P < .05 vs group 1 μL of PRP, ‡P < .05 vs group 10 μL of PRP for Figs A, B, and C. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

4 Fig 3 (A) Time course of ischemic/non-ischemic blood perfusion ratio in the different groups of mice with various treatments. (B) Ischemic/non-ischemic blood perfusion ratio in the different groups at four weeks after various treatments. (C) Blood perfusion ratio in contralateral nonischemic hind limb pre-treatment and four weeks after treatment of PRP-sr. Group C = Control, group PPP = treatment with 100 μL platelet-poor plasma, group PRP-sol = treatment with solution form of 100 μL platelet-rich plasma, group PRP-sr = treatment with sustained-release form of 100 μL platelet-rich plasma; n = 10 in each group. *P < .05 vs group C, †P < .05 vs group PPP. ‡P < .05 vs group PRP-sol. NS = no significant difference for groups pre-treatment and post-treatment with PRP-sr. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

5 Fig 4 Vascular density (A) and mature vessel density (B) in the ischemic calf muscles stained with anti-human von Willebrand factor (vWF) and anti-human smooth muscle actin (SMA) antibody at four weeks after surgery, respectively. Representative photomicrographs of the ischemic calf muscles double stained with the anti-human vWF and the anti-human SMA antibody at four weeks after surgery (C). Scale bars indicate 100 μm. Left column: vWF positive vessels; middle column: SMA positive vessels; right column: double stained vessels. Representative photomicrographs of the ischemic calf muscles single stained (Diaminobenzyn, DAB) with the anti-human vWF and the anti- human SMA antibody at four weeks after surgery (D). Scale bars indicate 100 μm. Left column: vWF positive vessels; right column: SMA positive vessels. Group C = Control, group PPP = treatment with 100 μL platelet-poor plasma, group PRP-sol = treatment with solution form of 100 μL platelet-rich plasma, group PRP-sr = treatment with sustained-release form of 100 μL platelet-rich plasma; n = 10 in each group. *P < .05 vs group C, †P < .05 vs group PPP. ‡P < .05 vs group PRP-sol. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

6 Fig 5 CD34+ cells in the peripheral blood and in the ischemic calf muscles at three days after surgery. Upper left panel: peripheral blood CD34+ cells detected by FACS; upper right panel: CD34+ cells in ischemic calf muscles stained with anti-mouse CD34 antibody. Lower panel: representative photomicrographs of the ischemic calf muscles stained with the anti- mouse CD34 antibody. Scale bars indicate 100 μm. Left column: GFP positive cell; middle column: CD34 positive cells; right Column: co-located cells. Group C = Control, group PRP-sr = treatment with sustained-release form of 100 μL platelets- rich plasma; n = 12 in each group. NS: not significant, *P < .05 vs group C. Journal of Vascular Surgery  , e2DOI: ( /j.jvs ) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

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