Objective
To evaluate the potential for biomimetic self-assembling fluorosurfactant polymer (FSP) coatings incorporating either heptamaltose (M7-FSP) to block non-specific protein adsorption, the cell adhesive RGD peptide (RGD-FSP), or the endothelial cell-selective CRRETAWAC peptide (cRRE-FSP) to improve patency and endothelialization in small diameter ePTFE vascular graft implants.
Methods
ePTFE vascular grafts (4 mm diameter, 5 cm length) were coated with either M7-FSP, RGD-FSP, or cRRE-FSP by dissolving FSPs in distilled water and flowing solution through the graft lumen for 24 h. Coatings were confirmed by receding water contact angle measurements on the lumen surface. RGD-FSP and cRRE-FSP grafts were pre-sodded in vitro with porcine pulmonary artery endothelial cells (PPAECs) using a custom-designed flow system. PPAEC coverage on the lumen surface was visualized with epifluorescent microscopy and quantified. Grafts were implanted as carotid artery interposition bypass grafts in 7 pigs for 33 ± 2 days (ePTFE n=3, M7-FSP n=4, RGD-FSP n=3, cRRE-FSP n=4). Patency was confirmed immediately after implant with duplex color-flow ultrasound and at explant with contrast-enhanced angiography. Grafts were sectioned for histology and stained: Movat’s pentachrome stain to outline vascular layers; immunofluorescent staining to identify endothelial cells (anti-von Willebrand factor antibody) and immunohistochemical staining to identify smooth muscle cells (anti-smooth muscle α-actin antibody). Neoinitima:lumen area ratio was determined to evaluate neointimal hyperplasia.
Results
Receding water contact angle measurements on graft luminal surfaces were significantly lower (P < .05) on FSP-coated ePTFE surfaces (M7-FSP: 40 ± 16°, RGD-FSP: 25 ± 10°, cRRE-FSP: 33 ± 16°) compared to uncoated ePTFE (126 ± 2°), confirming presence of the FSP layer. In vitro sodding of PPAECs on RGD-FSP and cRRE-FSP grafts resulted in a confluent monolayer of PPAECs on the luminal surface, with similar cell population on RGD-FSP (1200 ± 187 cells/mm2) and cRRE-FSP (1134 ± 153 cells/mm2) grafts. All grafts were patent immediately after implant, and 1/3 uncoated, 2/3 RGD-FSP, 2/4 M7-FSP, and 2/4 cRRE-FSP grafts remained patent after 1 month. PPAEC coverage of the lumen surface was seen in all patent grafts. RGD-FSP grafts had a slightly higher neointima:lumen area ratio (0.53 ± 0.06) compared to uncoated (0.29 ± 0.15), M7-FSP (0.20 ± 0.15), or cRRE-FSP (0.17 ± 0.09) grafts.
Conclusion
Biomimetic FSP coated ePTFE grafts can be utilized successfully in vivo and have potential to support endothelialization. Grafts modified with the M7-FSP and cRRE-FSP showed lower intimal hyperplasia compared to RGD-FSP grafts.