Effect of retroviral transduction on human endothelial cell phenotype and adhesion to Dacron vascular grafts Ronald J. Jankowski, MS, Donald A. Severyn,

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Presentation transcript:

Effect of retroviral transduction on human endothelial cell phenotype and adhesion to Dacron vascular grafts Ronald J. Jankowski, MS, Donald A. Severyn, MS, David A. Vorp, PhD, William R. Wagner, PhD Journal of Vascular Surgery Volume 26, Issue 4, Pages 676-684 (October 1997) DOI: 10.1016/S0741-5214(97)70069-7 Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Flow cytometric analysis of ICAM-1 and TF expression. ntEC and rtEC on coverslips were exposed to 2.5 dynes/cm2 wall shear stress for 6 hours or remained static. IL-1β (100 U/ml) was added to one half of the static cultures as positive control. All conditions were normalized to static ntEC cultures. Values shown represent normalized percentage of positive fluorescent cells for ICAM-1 (A), and TF (B). Data are expressed as mean + SEM. **p < 0.01 versus static ntEC. (Other significant differences of interest, but not noted on the graphs: A: rtEC w/shear vs ntEC w/shear, rtEC vs rtEC IL-β, p < 0.01; B: rtEC vs rtEC IL-1β, p < 0.01.) Journal of Vascular Surgery 1997 26, 676-684DOI: (10.1016/S0741-5214(97)70069-7) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Flow cytometric analysis of ICAM-1 and TF expression. ntEC and rtEC on coverslips were exposed to 2.5 dynes/cm2 wall shear stress for 6 hours or remained static. IL-1β (100 U/ml) was added to one half of the static cultures as positive control. All conditions were normalized to static ntEC cultures. Values shown represent normalized percentage of positive fluorescent cells for ICAM-1 (A), and TF (B). Data are expressed as mean + SEM. **p < 0.01 versus static ntEC. (Other significant differences of interest, but not noted on the graphs: A: rtEC w/shear vs ntEC w/shear, rtEC vs rtEC IL-β, p < 0.01; B: rtEC vs rtEC IL-1β, p < 0.01.) Journal of Vascular Surgery 1997 26, 676-684DOI: (10.1016/S0741-5214(97)70069-7) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Platelet (A) and PMN (B) adhesion to ntEC and rtEC monolayers. Confluent coverslips were perfused for 5 minutes at a wall shear stress of 2.5 dynes/cm2 for platelet adhesion data, and for 10 minutes at 2.5 dynes/cm2 for PMN adhesion data. % platelet coverage, percentage of total image area occupied by adherent platelets. Data are expressed as mean + SEM. **p < 0.01 versus ntEC. (Other significant differences of interest, but not noted on the graphs: A: rtEC vs rtEC PMA, p < 0.01; B: rtEC vs rtEC IL-1β, p < 0.01.) Journal of Vascular Surgery 1997 26, 676-684DOI: (10.1016/S0741-5214(97)70069-7) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Platelet (A) and PMN (B) adhesion to ntEC and rtEC monolayers. Confluent coverslips were perfused for 5 minutes at a wall shear stress of 2.5 dynes/cm2 for platelet adhesion data, and for 10 minutes at 2.5 dynes/cm2 for PMN adhesion data. % platelet coverage, percentage of total image area occupied by adherent platelets. Data are expressed as mean + SEM. **p < 0.01 versus ntEC. (Other significant differences of interest, but not noted on the graphs: A: rtEC vs rtEC PMA, p < 0.01; B: rtEC vs rtEC IL-1β, p < 0.01.) Journal of Vascular Surgery 1997 26, 676-684DOI: (10.1016/S0741-5214(97)70069-7) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Flow cytometric analysis of ICAM-1 and TF expression on seeded MVECs. ntEC-seeded and rtEC-seeded Dacron grafts were exposed to 2.5 dynes/cm2 wall shear stress for 6 hours or remained static. All conditions were normalized to static ntECs on coverslips (ntEC control). IL-1β (100 U/ml) was added to one half of the static cultures as positive control. Graph values indicate normalized percentage of fluorescent cells positive for ICAM-1 (A) and TF (B) expression. Data are expressed as mean + SEM. **p < 0.01; *p < 0.05 versus static ntEC control. (Other significant differences of interest, but not noted on the graphs: B: ntEC w/shear vs rtEC w/shear, p < 0.05; ntEC vs ntEC w/shear, p < 0.01.) Journal of Vascular Surgery 1997 26, 676-684DOI: (10.1016/S0741-5214(97)70069-7) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Flow cytometric analysis of ICAM-1 and TF expression on seeded MVECs. ntEC-seeded and rtEC-seeded Dacron grafts were exposed to 2.5 dynes/cm2 wall shear stress for 6 hours or remained static. All conditions were normalized to static ntECs on coverslips (ntEC control). IL-1β (100 U/ml) was added to one half of the static cultures as positive control. Graph values indicate normalized percentage of fluorescent cells positive for ICAM-1 (A) and TF (B) expression. Data are expressed as mean + SEM. **p < 0.01; *p < 0.05 versus static ntEC control. (Other significant differences of interest, but not noted on the graphs: B: ntEC w/shear vs rtEC w/shear, p < 0.05; ntEC vs ntEC w/shear, p < 0.01.) Journal of Vascular Surgery 1997 26, 676-684DOI: (10.1016/S0741-5214(97)70069-7) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 MVEC retention after 6 hour pulsatile perfusions at 2.5 dynes/cm2. Data are expressed as the total percentage of viable ECs remaining on graft surface after perfusion, as compared with cell numbers before flow exposure. Two graft segments from each perfused graft were evaluated for cell retention, one proximal and one distal with respect to direction of perfusate flow. Journal of Vascular Surgery 1997 26, 676-684DOI: (10.1016/S0741-5214(97)70069-7) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions