In vivo evaluation of biomimetic fluorosurfactant polymer-coated expanded polytetrafluoroethylene vascular grafts in a porcine carotid artery bypass model.
In vivo evaluation of biomimetic fluorosurfactant polymer-coated expanded polytetrafluoroethylene vascular grafts in a porcine carotid artery bypass model.
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DOI：
10.1016/j.jvs.2015.01.060
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发表时间：
2016-06
影响因子：
4.3
通讯作者：
Kottke-Marchant K
中科院分区：
文献类型：
--
作者：
Bastijanic JM;Marchant RE;Kligman F;Allemang MT;Lakin RO;Kendrick D;Kashyap VS;Kottke-Marchant K
评价仿生自组装含氟表面活性剂聚合物（FSP）涂层结合七麦芽糖（M7-FSP）以阻断非特异性蛋白质吸附、细胞粘附性RGD肽（RGD-FSP）或内皮细胞选择性CRRETAWAC肽（cRRE-FSP）以改善小直径ePTFE血管移植物植入物的通畅性和内皮化的潜力。通过将FSP溶解在蒸馏水中并使溶液流过移植物管腔24小时，用M7-FSP、RGD-FSP或cRRE-FSP涂覆ePTFE血管移植物（4 mm直径，5cm长）。涂层通过测量管腔表面上的后退水接触角来确认。使用定制设计的流动系统在体外用猪肺动脉内皮细胞（PPAEC）预接种RGD-FSP和cRRE-FSP移植物。用落射荧光显微镜观察管腔表面上的PPAEC覆盖并定量。将移植物作为颈动脉间置旁路移植物植入7只猪中，持续33 ± 2天（ePTFE n=3，M7-FSP n=4，RGD-FSP n=3，cRRE-FSP n=4）。植入后立即用双功彩色血流超声和取出时用造影剂增强血管造影证实通畅。对移植物进行组织学切片和染色：Movat五色染色以勾勒血管层;免疫荧光染色以识别内皮细胞（抗血管性血友病因子抗体）和免疫组织化学染色以识别平滑肌细胞（抗平滑肌α-肌动蛋白抗体）。测定新生内膜：管腔面积比以评价新生内膜增生。FSP涂层ePTFE表面（M7-FSP：40 ± 16°，RGD-FSP：25 ± 10°，cRRE-FSP：33 ± 16°）的移植物管腔表面的后退水接触角测量值显著低于（P <0.05）未涂层ePTFE（126 ± 2°），证实存在FSP层。PPAEC在RGD-FSP和cRRE-FSP移植物上的体外sodding导致在管腔表面上的PPAEC的汇合单层，在RGD-FSP（1200 ± 187个细胞/mm 2）和cRRE-FSP（1134 ± 153个细胞/mm 2）移植物上具有相似的细胞群。所有移植物在植入后立即通畅，1/3无涂层、2/3 RGD-FSP、2/4 M7-FSP和2/4 cRRE-FSP移植物在1个月后保持通畅。在所有未闭移植物中均观察到管腔表面的PPAEC覆盖。与无涂层（0.29 ± 0.15）、M7-FSP（0.20 ± 0.15）或cRRE-FSP（0.17 ± 0.09）移植物相比，RGD-FSP移植物的新生内膜：管腔面积比（0.53 ± 0.06）略高。仿生FSP涂层的ePTFE移植物可以成功地在体内使用，并具有支持内皮化的潜力。与RGD-FSP移植物相比，用M7-FSP和cRRE-FSP修饰的移植物显示出较低的内膜增生。
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. 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. 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. 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.