Randall Adolph, BS, David A. Vorp, PhD, David L. Steed, MD, Marshall W

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Presentation transcript:

Cellular content and permeability of intraluminal thrombus in abdominal aortic aneurysm Randall Adolph, BS, David A. Vorp, PhD, David L. Steed, MD, Marshall W. Webster, MD, Marina V. Kameneva, PhD, Simon C. Watkins, PhD Journal of Vascular Surgery Volume 25, Issue 5, Pages 916-926 (May 1997) DOI: 10.1016/S0741-5214(97)70223-4 Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Hematoxylin and eosin–stained section of a representative thrombus specimen. It appears as though thrombus has within its structure spaces that may constitute interconnected canaliculi. It also qualitatively appears that areas of observed canaliculi increase with distance from luminal aspect of thrombus. Original magnification, 100×. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 A, Lipid-filled macrophage is at leading edge of canaliculus (original magnification, 2500×). B, Within the depth of thrombus, lipid-rich macrophages were abundant. It appears that macrophage is phagocytosing a normal erythrocyte (arrow) (original magnification, 2500×). C, Scanning electron micrograph of Mercoxy resin cast. Before polymerizing, resin entered thrombus via gravitational force. C is a low-power micrograph of a representative thrombus specimen. D, Higher-power micrograph from same specimens. It is evident from these micrographs that there is a reticular nature to penetrating resin that may represent canaliculi that penetrate thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 A, Lipid-filled macrophage is at leading edge of canaliculus (original magnification, 2500×). B, Within the depth of thrombus, lipid-rich macrophages were abundant. It appears that macrophage is phagocytosing a normal erythrocyte (arrow) (original magnification, 2500×). C, Scanning electron micrograph of Mercoxy resin cast. Before polymerizing, resin entered thrombus via gravitational force. C is a low-power micrograph of a representative thrombus specimen. D, Higher-power micrograph from same specimens. It is evident from these micrographs that there is a reticular nature to penetrating resin that may represent canaliculi that penetrate thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 A, Lipid-filled macrophage is at leading edge of canaliculus (original magnification, 2500×). B, Within the depth of thrombus, lipid-rich macrophages were abundant. It appears that macrophage is phagocytosing a normal erythrocyte (arrow) (original magnification, 2500×). C, Scanning electron micrograph of Mercoxy resin cast. Before polymerizing, resin entered thrombus via gravitational force. C is a low-power micrograph of a representative thrombus specimen. D, Higher-power micrograph from same specimens. It is evident from these micrographs that there is a reticular nature to penetrating resin that may represent canaliculi that penetrate thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 A, Lipid-filled macrophage is at leading edge of canaliculus (original magnification, 2500×). B, Within the depth of thrombus, lipid-rich macrophages were abundant. It appears that macrophage is phagocytosing a normal erythrocyte (arrow) (original magnification, 2500×). C, Scanning electron micrograph of Mercoxy resin cast. Before polymerizing, resin entered thrombus via gravitational force. C is a low-power micrograph of a representative thrombus specimen. D, Higher-power micrograph from same specimens. It is evident from these micrographs that there is a reticular nature to penetrating resin that may represent canaliculi that penetrate thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Immunohistochemical analysis was used to quantify the number of macrophages present at various depths within thrombus. 0 mm refers to luminal surface. This representative thrombus specimen shows a thrombic specimen stained with a fluorescent conjugated rat antihuman mAb directed against human macrophages and shows general trend that was observed. On quantification and statistical analysis (Fig. 4), it was seen that mean and median number of macrophages decreased with increasing depth toward luminal surface of thrombus. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Data from all 24 thrombi were combined and mean number of macrophages quantified. A, Histogram representing mean number of macrophages present at various defined depths within thrombus. Correlation of decreasing number of macrophages with increasing depth was observed. B, To determine whether this correlation was statistically significant, a statistical regression analysis was performed. This statistical test reveals that a significant trend does exist; with increasing depth toward abluminal aspect of thrombus there are significantly fewer macrophages than are present at luminal surface. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Data from all 24 thrombi were combined and mean number of macrophages quantified. A, Histogram representing mean number of macrophages present at various defined depths within thrombus. Correlation of decreasing number of macrophages with increasing depth was observed. B, To determine whether this correlation was statistically significant, a statistical regression analysis was performed. This statistical test reveals that a significant trend does exist; with increasing depth toward abluminal aspect of thrombus there are significantly fewer macrophages than are present at luminal surface. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Sample Western blots show relative amounts of fibrin deposition and degradation at defined depths into thrombus. A, Labeling for fibrin degradation products in two separate specimens. Samples were taken at 0, 2, and 4 mm within the thrombus and gels equivalently loaded. It can be seen that band intensity increases as distance increases. This indicates that fibrin degradation increases with depth into thrombus. This trend is observed in each thrombi examined. B, Abundance of GpIIbIIIa, in the same two separate thrombi. Abundance decreases with depth into thrombus. Numbers above lanes indicate distance, in millimeters, from luminal surface of thrombus. Band representing fibrin degradation is seen at 150 kD, whereas band representing fibrin deposition is seen at 133 kD. Journal of Vascular Surgery 1997 25, 916-926DOI: (10.1016/S0741-5214(97)70223-4) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions