Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, Nebraska, United States of America.
Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska, United States of America.
PLoS One. 2018 Jul 2;13(7):e0199925. doi: 10.1371/journal.pone.0199925. eCollection 2018.
In this study, heparin-loaded poly-ɛ-caprolactone (PCL) fibrous mats were prepared and characterized based on their physical, cytotoxic, thermal, and biological properties. The main objective of the work described here was to test the hypothesis that incorporation of heparin into a PCL carrier could serve as bio-compatible material capable of inhibiting Human Papillomavirus (HPV) infection. The idea of firmly anchoring heparin to capture soluble virus, vs. a slow heparin release to inhibit a virus in solution was tested. Thus, one material was produced via conventional heparin matrix encapsulation and electrohydrodynamic fiber processing in one step. A second type of material was obtained via heparin crosslinking. This was achieved by running a carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling reaction on preformed PCL fibers. In vitro HPV16 L1 protein binding capacity studies were performed. Infectivity assays were done using HPV16 pseudoviruses (PsVs) carrying a GFP plasmid to directly test the ability of the fibrous mats to prevent internalization of HPV PsVs. The crosslinked heparin material presented a dissociation constant (Kd) value comparable to those found in the literature for different heparin-protein L1 peptide interactions. Both materials significantly reduced internalization of HPV PsVs, with a reduction of 94% of PsVs internalization when matrix encapsulated heparin-loaded material was present. Differences in performance between the two proposed structures are discussed.
在这项研究中,制备了载肝素的聚己内酯(PCL)纤维垫,并基于其物理、细胞毒性、热和生物学性质对其进行了表征。这里描述的工作的主要目的是检验以下假设,即将肝素掺入 PCL 载体中可以作为生物相容材料,能够抑制人乳头瘤病毒(HPV)感染。这里测试了将肝素牢固固定以捕获可溶性病毒的想法,而不是通过缓慢释放肝素来抑制溶液中的病毒。因此,一种材料是通过传统的肝素基质包封和一步电纺纤维加工生产的。另一种材料是通过肝素交联获得的。这是通过在预成型的 PCL 纤维上进行碳二亚胺/N-羟基琥珀酰亚胺(EDC/NHS)偶联反应来实现的。进行了 HPV16 L1 蛋白结合容量研究。使用携带 GFP 质粒的 HPV16 假病毒进行感染性测定,以直接测试纤维垫阻止 HPV PsV 内化的能力。交联肝素材料的解离常数(Kd)值与文献中不同肝素-蛋白 L1 肽相互作用的 Kd 值相当。两种材料均显著降低了 HPV PsV 的内化,当存在基质包封的载肝素材料时,HPV PsV 的内化减少了 94%。讨论了两种所提出结构之间的性能差异。