Decellularized heart valve as a scaffold for in vivo recellularization: Deleterious effects of granulocyte colony-stimulating factor Francis Juthier,

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Presentation transcript:

Decellularized heart valve as a scaffold for in vivo recellularization: Deleterious effects of granulocyte colony-stimulating factor Francis Juthier, MD, André Vincentelli, MD, PhD, Julien Gaudric, MD, Delphine Corseaux, PhD, Olivier Fouquet, MD, Christine Calet, MD, Thierry Le Tourneau, MD, PhD, Valérie Soenen, BS, Christophe Zawadzki, BS, Olivier Fabre, MD, Sophie Susen, MD, Alain Prat, MD, Brigitte Jude, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 131, Issue 4, Pages 843-852 (April 2006) DOI: 10.1016/j.jtcvs.2005.11.037 Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Kinetics of white blood cell counts in lambs after subcutaneous injection of recombinant human G-CSF (10 μg/kg) for 7 days. The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Results of the decellularization process (hematoxylin, eosin, and safran staining). Before and after decellularization: A, native valve conduit; B, leaflet. A complete decellularization of the aortic wall and of the leaflets without destruction of the fibrous structure is shown. The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 A and B, Representative aspects of normal aortography (A) at 16 weeks and perioperative aspect of the graft in the thoracic descending aorta (B). C and D, Specific aspect of the 16 weeks after implantation aortography (C) and of the explanted valve (D) in the animal that underwent complete valve thrombosis (xenograft group). Arrows indicate site of the graft implantation. The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Macroscopic examination at 16 weeks. A, Control group; B, G-CSF group; C, Xenograft group. In the xenograft and the G-CSF groups, but not in the control group, gross observation showed calcifications of the leaflets and of the aortic wall, leaflet thickening, and shrinkage at 16 weeks. The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 Representative histologic aspects in the grafted aortic wall at 16 weeks in the 3 groups (top row, control group; middle row, G-CSF group; bottom row, xenograft group). A, Adventitia (Masson trichrome staining): collagen fibers and limited infiltration by inflammatory cells were visible in the control group. Inflammatory cells were more prominent in the G-CSF and xenograft groups than in the control group. B (anti-VWF antibody), Adventitial neovascularization (arrows) was limited in the control group and abundant in the G-CSF and xenograft groups. C (Masson trichrome staining), Neointimal proliferation tended to be more important in the G-CSF and xenograft groups than in the control group. D (anti-α-actin staining), Evidence of strong α-actin staining in the neointimal layer in the 3 groups. The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 6 Orcein, von Kossa, and immunochemical staining with anti-VWF antibody in the 3 groups and representative aspects of the leaflets are shown (top row, control group; middle row, G-CSF group; bottom row, xenograft group). A (orcein staining), Elastic fibers remained parallel in the control group and were disrupted and disorganized in the xenograft and G-CSF groups. B (Von Kossa staining), Evidence of heavy calcifications in the aortic wall in the G-CSF and xenograft groups and not in the control group. C (anti-VWF antibody), The luminal surface of the aortic wall was covered by a continuous layer of endothelial cells in the control group and by a disrupted layer (arrows) in the G-CSF and xenograft groups. D (Masson trichrome staining), Representative aspects of the leaflets in the 3 groups, with calcifications in the G-CSF and xenograft groups. The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 7 Time course of the intima/media ratio in the 3 groups. The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions

Figure 8 Results of fluorescence in situ hybridization on the cells eluted from the graft (A), a positive control on ovine blood (B), and a negative control on porcine blood (C). The Journal of Thoracic and Cardiovascular Surgery 2006 131, 843-852DOI: (10.1016/j.jtcvs.2005.11.037) Copyright © 2006 The American Association for Thoracic Surgery Terms and Conditions