Endothelial seeding of polytetrafluoroethylene popliteal bypasses

Herring, Malcolm B.; Compton, Rebecca S.; LeGrand, Daniel R.; Gardner, Austin; Madison, David; Glover, John L.

doi:10.1067/mva.1987.avs0060114

Cited by 75 publications

(19 citation statements)

References 8 publications

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“…Both clinical as well as basic studies have shown that ePTFE performs inferior to veins when used as bypass grafts [1, 2]. This led to the concept of cell seeding in which autologous endothelial cells are transplanted on ePTFE and so form an anticoagulant layer between blood and prosthesis [29, 30, 31]. Good results with cell seeding of grafts have been reported using HAEC [8].…”

Section: Discussionmentioning

confidence: 94%

Thrombomodulin Activity of Fat-Derived Microvascular Endothelial Cells Seeded on Expanded Polytetrafluorethylene

Joosten¹,

Verhagen²,

Heijnen-Snyder³

et al. 1999

J Vasc Res

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Lining the luminal surface of prosthetic vascular grafts with endothelial cells (cell seeding) will lower its thrombogenicity. Commonly used macrovascular human adult endothelial cells (HAEC) require in vitro cultivation before large enough numbers are obtained to cover grafts confluently. Fat-derived microvascular endothelial cells (MVEC) prove to be a good alternative as they can be harvested in much larger numbers while showing similar antithrombotic and fibrinolytic characteristics. An important anticoagulant function of macrovascular endothelial cells is due to the activity of thrombomodulin (TM) on their surface. In this study, the presence and functional activity of TM on fat-derived microvascular cells used in cell seeding was investigated. The expression and localization of TM on MVEC was studied using immunohistochemistry. Functional activity of TM on MVEC was measured by the generation of activated protein C (APC) and was compared to human umbilical vein endothelial cells (HUVEC). TM activity was studied in MVEC seeded on expanded polytetrafluorethylene (ePTFE) vascular prostheses and compared to blank prostheses. We found that TM was expressed on the surface of MVEC, both in vivo and vitro. TM-dependent generation of APC differed significantly between MVEC and HUVEC (3.98 ± 1.2 vs. 3.0 ± 0.7 nM, respectively). After seeding MVEC on vascular prostheses, TM activity did not change. APC generation was significantly higher on MVEC-seeded vascular grafts compared to blank grafts (4.0 ± 0.7 vs. 1.7 ± 0.5 nM, respectively). We conclude that TM is present and highly active on cultured MVEC. When seeded on ePTFE, MVEC retain the possibility to inhibit thrombin coagulant activity and to activate protein C. Therefore, since MVEC are readily available, the anticoagulant properties demonstrated here indicate that this cell type is suitable for cell seeding of vascular prostheses.

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Section: Discussionmentioning

confidence: 94%

Thrombomodulin Activity of Fat-Derived Microvascular Endothelial Cells Seeded on Expanded Polytetrafluorethylene

Joosten¹,

Verhagen²,

Heijnen-Snyder³

et al. 1999

J Vasc Res

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“…This has been the same technique used in determining the nec essary inoculum density and in our previous seeding experiments [2,6,16]. The control veins in group B were bathed and distended in DME without papaverine in order to dif ferentiate the effects of the papaverine from any effects of the use of tissue culture me dium.…”

Section: Discussionmentioning

confidence: 99%

“…Venous endothelial cells have been used in many experiments to provide a nonthrombogenic interface between the pros thetic graft and the blood [1,2]. Established methods of harvesting include both mechan ical and enzymatic techniques.…”

Section: Introductionmentioning

confidence: 99%

Effect of Papaverine on Endothelial Cell Harvest from Canine External Jugular Veins

Le²,

Fietsam³

et al. 1991

J Vasc Res

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Endothelial cell seeding procedures have been developed to line prosthetic bypass grafts used in peripheral vascular disease; however, because of current inefficient cell harvest techniques a high ratio of vein-to-graft area is necessary. This study was done to determine if the use of papaverine, a smooth muscle cell relaxant, would affect the number or viability of endothelial cells harvested from canine external jugular veins. Using a 0.12 mg/ml solution of papaverine in tissue culture medium to bathe the veins during disscetion and excision, the viable cell yield was 2.20 ± 1.16 (cells × 10⁴/cm²). A control group of veins using standard dissection technique gave a yield of 0.97 ± 0.40 (p = 0.025). A second group of veins dissected while bathed in tissue culture medium alone gave a yield of 1.82 ± 0.75, compared to a yield of 2.73 ± 0.45 for papaverine harvested veins (p = 0.009). Percent cell viability was not significantly different for any of the groups: 73, 70, and 76% for papaverine, control and media only veins, respectively. The papaverine-harvested cells and those harvested with medium alone grew to 95% confluence in tissue culture in 9.8 ± 1.1 and 9.9 ± 0.9 days, respectively. Compared to conventional surgical techniques, use of papaverine more than doubled the endothelial cell yield from excised vein segments without adversely affecting viability or rate of growth in cell culture.

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“…Although improved patency has been observed in some reports, clinical use and results have remained controversial and somewhat disappointing. [7][8][9][10][11] In addition, autologous EC-seeded grafts have several clinical limitations. They not only require previous EC harvest but also an incubation period, making them available to a patient only on a semi-elective basis.…”

Section: Introductionmentioning

confidence: 99%

A biologically active VEGF construct in vitro: Implications for bioengineering‐improved prosthetic vascular grafts

Stone

Phaneuf

Sivamurthy

et al. 2001

J. Biomed. Mater. Res.

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Prosthetic arterial grafts are unable to develop an intact endothelial lining after implantation, predisposing them to fail. Strategies have been sought to enhance endothelialization using growth factors and cytokines. This study assessed the biologic activity of vascular endothelial growth factor (VEGF) covalently linked to bovine serum albumin (BSA). Native and modified VEGF were assayed for endothelial cell migration and proliferation. Migration assays were performed comparing the effects of 2% fetal bovine serum (FBS), 50 ng/mL, 100 ng/mL, and 200 ng/mL of native VEGF and VEGF-BSA. Proliferation assays were performed by using Alamar Blue comparing cellular growth in 1% FBS, 10% FBS, 100 ng/mL unbound VEGF, and 100 ng/mL VEGF-BSA. VEGF is a potent chemotactic agent for endothelial cells in both unbound and bound states. Native VEGF solutions (50 ng/mL, 100 ng/mL, and 200 ng/mL) stimulated 23.9 cells/high power field (HPF), 35.3 cells/HPF, and 49.1 cells/HPF (p < 0.005). VEGF-BSA solutions stimulated 25.9 cells/HPF, 39.1 cells/HPF, and 69.0 cells/HPF (p < 0.001). VEGF-BSA and native VEGF supported similar increased cellular proliferation compared with 1% FBS media (p < 0.002). Modified VEGF retains its chemotactic and proliferative properties in vitro. These findings suggest that bare prosthetic surfaces lined with VEGF bound to a "basecoat" albumin may support endothelial cell proliferation and migration and thereby offer new strategies to improve graft patency.

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Endothelial seeding of polytetrafluoroethylene popliteal bypasses

Cited by 75 publications

References 8 publications

Thrombomodulin Activity of Fat-Derived Microvascular Endothelial Cells Seeded on Expanded Polytetrafluorethylene

Thrombomodulin Activity of Fat-Derived Microvascular Endothelial Cells Seeded on Expanded Polytetrafluorethylene

Effect of Papaverine on Endothelial Cell Harvest from Canine External Jugular Veins

A biologically active VEGF construct in vitro: Implications for bioengineering‐improved prosthetic vascular grafts

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