Chen Lei, He Haipeng, Wang Mian, Li Xiaoxi, Yin Henghui
1Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510080 China.
2Department of Vascular Surgery, The First Affiliated Hospital of Ji'nan University, Guangzhou, 510630 China.
Tissue Eng Regen Med. 2017 Mar 2;14(4):359-370. doi: 10.1007/s13770-017-0044-3. eCollection 2017 Aug.
Expanded polytetrafluoroethylene (ePTFE) polymers do not support endothelialization because of nonconductive characteristics towards cellular attachment. Inner surface modification of the grafts can improve endothelialization and increase the long-term patency rate of the ePTFE vascular grafts. Here we reported a method of inner-surface modification of ePTFE vascular graft with extracellular matrix (ECM) and CD34 monoclonal antibodies (CD34 mAb) to stimulate the adhesion and proliferation of circulating endothelial progenitor cells on ePTFE graft to enhance graft endothelialization. The inner surface of ECM-coated ePTFE grafts were linked with CD34 mAb in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/-hydroxysuccinimide (EDC/NHS) solution and the physicochemical properties, surface morphology, biocompatibility, and hemocompatibility of the grafts were studied. The hydrophilicity of CD34 mAb-coated graft inner surface was significantly improved. Fourier transform infrared spectroscopy analysis confirmed ECM and CD34 mAb cross-linking in the ePTFE vascular grafts with our method. Scanning electron microscopy analysis showed protein layer covering uniformly on the inner surface of the modified grafts. The cell-counting kit-8 (CCK-8) assay confirmed that the modified graft has no obvious cytotoxicity. The modified graft showed a low hemolytic rate (0.9%) in the direct contact hemolysis test, suggesting the modification improved hemocompatibility of biopolymers. The modification also decreased adhesion of platelets, while significantly increased the adhesion of endothelial cells on the grafts. We conclude that our method enables ePTFE polymers modification with ECM and CD34 mAb, facilitates endothelialization, and inhibits platelet adhesion on the grafts, thus may increase the long-term patency rate of the prosthetic bypass grafts.
膨体聚四氟乙烯(ePTFE)聚合物由于对细胞附着具有非传导性特征,不支持内皮化。移植物的内表面改性可以改善内皮化并提高ePTFE血管移植物的长期通畅率。在此,我们报道了一种用细胞外基质(ECM)和CD34单克隆抗体(CD34 mAb)对ePTFE血管移植物进行内表面改性的方法,以刺激循环内皮祖细胞在ePTFE移植物上的黏附和增殖,从而增强移植物内皮化。在1-乙基-3-(3-二甲基氨基丙基)碳二亚胺/ N-羟基琥珀酰亚胺(EDC / NHS)溶液存在的情况下,将ECM包被的ePTFE移植物的内表面与CD34 mAb连接,并研究了移植物的物理化学性质、表面形态、生物相容性和血液相容性。CD34 mAb包被的移植物内表面的亲水性得到显著改善。傅里叶变换红外光谱分析证实了我们的方法使ECM和CD34 mAb在ePTFE血管移植物中发生交联。扫描电子显微镜分析显示蛋白质层均匀覆盖在改性移植物的内表面。细胞计数试剂盒-8(CCK-8)检测证实改性移植物没有明显的细胞毒性。改性移植物在直接接触溶血试验中显示出较低的溶血率(0.9%),表明该改性提高了生物聚合物的血液相容性。该改性还减少了血小板的黏附,同时显著增加了内皮细胞在移植物上的黏附。我们得出结论,我们的方法能够用ECM和CD34 mAb对ePTFE聚合物进行改性,促进内皮化,并抑制血小板在移植物上的黏附,从而可能提高人工旁路移植物的长期通畅率。