Influence of vein valves in the development of arteriosclerosis in venoarterial grafts in the rabbit  Aurelio Chaux, MDa (by invitation), Xin Min Ruan, MDa (by invitation), Michael C. Fishbein, MDb (by invitation), Meenu Sandhu, MSc (by invitation), Jack M. Matloff, MDa  The Journal of Thoracic and Cardiovascular Surgery  Volume 110, Issue 5, Pages 1381-1390 (November 1995) DOI: 10.1016/S0022-5223(95)70061-7 Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 1 A, Cell density (number of cells per square micrometer) was largest at 2 weeks and progressively decreased during the time of the experiment. B, PCNA expression reached a maximum at 2 weeks, and it was minimal or nonexistent thereafter. Nodifference was detected between the two groups. The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 2 Media: Photomicrographs at a standard original magnification of 50× showing progressive increase in medial thicknessof veins with time that occurred in both treatment groups. A, Control; B, 2 weeks; C, 4 weeks; D, 6 weeks, E and F, 8 weeks. Note in E that at 8 weeks the media was sometimes more fibrous andless cellular and, as shown in F, sometimes contained numerous foamy macrophages (asterisks). a, adventitia; m, media. (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 2 Media: Photomicrographs at a standard original magnification of 50× showing progressive increase in medial thicknessof veins with time that occurred in both treatment groups. A, Control; B, 2 weeks; C, 4 weeks; D, 6 weeks, E and F, 8 weeks. Note in E that at 8 weeks the media was sometimes more fibrous andless cellular and, as shown in F, sometimes contained numerous foamy macrophages (asterisks). a, adventitia; m, media. (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 2 Media: Photomicrographs at a standard original magnification of 50× showing progressive increase in medial thicknessof veins with time that occurred in both treatment groups. A, Control; B, 2 weeks; C, 4 weeks; D, 6 weeks, E and F, 8 weeks. Note in E that at 8 weeks the media was sometimes more fibrous andless cellular and, as shown in F, sometimes contained numerous foamy macrophages (asterisks). a, adventitia; m, media. (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 2 Media: Photomicrographs at a standard original magnification of 50× showing progressive increase in medial thicknessof veins with time that occurred in both treatment groups. A, Control; B, 2 weeks; C, 4 weeks; D, 6 weeks, E and F, 8 weeks. Note in E that at 8 weeks the media was sometimes more fibrous andless cellular and, as shown in F, sometimes contained numerous foamy macrophages (asterisks). a, adventitia; m, media. (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 2 Media: Photomicrographs at a standard original magnification of 50× showing progressive increase in medial thicknessof veins with time that occurred in both treatment groups. A, Control; B, 2 weeks; C, 4 weeks; D, 6 weeks, E and F, 8 weeks. Note in E that at 8 weeks the media was sometimes more fibrous andless cellular and, as shown in F, sometimes contained numerous foamy macrophages (asterisks). a, adventitia; m, media. (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 2 Media: Photomicrographs at a standard original magnification of 50× showing progressive increase in medial thicknessof veins with time that occurred in both treatment groups. A, Control; B, 2 weeks; C, 4 weeks; D, 6 weeks, E and F, 8 weeks. Note in E that at 8 weeks the media was sometimes more fibrous andless cellular and, as shown in F, sometimes contained numerous foamy macrophages (asterisks). a, adventitia; m, media. (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 3 Valve: Photomicrographs demonstrating venous valvular changes. A, B, and C are normal valves (open arrows) at 10×, 25×, and 50× magnification; D, E, and F are valves from veins implanted for 2 to 6 weeks, shown at 10×, 25×, and 50× magnification. Note the marked thickening of the valve base. In E this consistsprimarily of smooth muscle cells. In F foamy macrophages (asterisks) are also present. D, E, and F also demonstrate medial proliferation of the adjacent venous wall (small arrows). (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 3 Valve: Photomicrographs demonstrating venous valvular changes. A, B, and C are normal valves (open arrows) at 10×, 25×, and 50× magnification; D, E, and F are valves from veins implanted for 2 to 6 weeks, shown at 10×, 25×, and 50× magnification. Note the marked thickening of the valve base. In E this consistsprimarily of smooth muscle cells. In F foamy macrophages (asterisks) are also present. D, E, and F also demonstrate medial proliferation of the adjacent venous wall (small arrows). (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 3 Valve: Photomicrographs demonstrating venous valvular changes. A, B, and C are normal valves (open arrows) at 10×, 25×, and 50× magnification; D, E, and F are valves from veins implanted for 2 to 6 weeks, shown at 10×, 25×, and 50× magnification. Note the marked thickening of the valve base. In E this consistsprimarily of smooth muscle cells. In F foamy macrophages (asterisks) are also present. D, E, and F also demonstrate medial proliferation of the adjacent venous wall (small arrows). (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 3 Valve: Photomicrographs demonstrating venous valvular changes. A, B, and C are normal valves (open arrows) at 10×, 25×, and 50× magnification; D, E, and F are valves from veins implanted for 2 to 6 weeks, shown at 10×, 25×, and 50× magnification. Note the marked thickening of the valve base. In E this consistsprimarily of smooth muscle cells. In F foamy macrophages (asterisks) are also present. D, E, and F also demonstrate medial proliferation of the adjacent venous wall (small arrows). (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 3 Valve: Photomicrographs demonstrating venous valvular changes. A, B, and C are normal valves (open arrows) at 10×, 25×, and 50× magnification; D, E, and F are valves from veins implanted for 2 to 6 weeks, shown at 10×, 25×, and 50× magnification. Note the marked thickening of the valve base. In E this consistsprimarily of smooth muscle cells. In F foamy macrophages (asterisks) are also present. D, E, and F also demonstrate medial proliferation of the adjacent venous wall (small arrows). (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 3 Valve: Photomicrographs demonstrating venous valvular changes. A, B, and C are normal valves (open arrows) at 10×, 25×, and 50× magnification; D, E, and F are valves from veins implanted for 2 to 6 weeks, shown at 10×, 25×, and 50× magnification. Note the marked thickening of the valve base. In E this consistsprimarily of smooth muscle cells. In F foamy macrophages (asterisks) are also present. D, E, and F also demonstrate medial proliferation of the adjacent venous wall (small arrows). (Hematoxylin and eosin stain.) The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions

Fig. 4 Immunohistochemistry: Photomicrographs showing results of immunohistochemical studies. A, Hematoxylin and eosin stain of valvular proliferation at 3 days (50×). B, PCNA staining showing numerous positive brown-staining cells (arrows). Inset shows positive control consisting of intestinal crypt epithelium (50×). Adjacent media had little PCNA positivity. C, Valve at 4 weeks showing no PCNA positivity. D, Example of vein wall proliferation of cells (PCNA positivity indicated by arrows) observed within the first 2 weeks after implantation (100×). E, An example of staining of the venous wall at 2 weeks for smooth muscle cell(s), specific act inindicating that virtually all the cells are smooth muscle cells (10×). F, a vein at 8 weeks, with primarily foam cells in the wall (hematoxylin and eosin stain, 10×). G, Most foam cells are negative for smooth muscle-specific actin(s) (100×). H, Another vein showing that foam cells stain positively with amacrophage-specific antibody (red), whereas smooth muscle cell(s) fail to stain (100×). The Journal of Thoracic and Cardiovascular Surgery 1995 110, 1381-1390DOI: (10.1016/S0022-5223(95)70061-7) Copyright © 1995 Mosby, Inc. Terms and Conditions