Structure and Function of Trypsin-Loaded Fibrinolytic Liposomes
Structure and Function of Trypsin-Loaded Fibrinolytic Liposomes
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DOI：
10.1155/2017/5130495
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Kolev, Krasimir
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文献类型：
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作者：
Tanka-Salamon, Anna;Bota, Attila;Kolev, Krasimir
蛋白酶包裹和其在血栓中的靶向释放可能有助于减少溶栓的出血并发症。我们的目的是制备空间稳定的胰蛋白酶脂质体（SSLT），并表征其结构和纤溶效率。制备了氢化大豆磷脂酰胆碱（SSLT），并利用透射电镜结合冷冻断裂（FF-TEM）、傅里叶变换红外光谱（FT-IR）和小角X射线散射（SAXS）对其结构进行了研究。在45，37，或24（omicron）C下，用比浊法和渗透驱动的裂解试验对纤维蛋白或血浆凝块进行纤溶活性检查。胰蛋白酶被证明附着在靠近脂质亲水/疏水性界面（FT-IR）的囊泡内表面（SAXS和FF-TEM）。SSLT的热敏性通过在45（omicron）C下增强的纤维蛋白溶解来证明：与37（omicron）C相比，将最大浊度降低至20%的时间减少了8.6%，并且在渗透溶解测定中纤维蛋白降解产物浓度比在24（omicron）C下高2倍至5倍。SSLT在静态和动态条件下对纤维蛋白凝块发挥其纤维蛋白溶解作用，而血浆凝块溶解仅在渗透驱动试验中观察到。SSLT在动态条件下改善的纤溶效率表明，它们可以作为溶解血管内血栓的新型治疗候选物，血管内血栓通常暴露于渗透力。
Protease encapsulation and its targeted release in thrombi may contribute to the reduction of haemorrhagic complications of thrombolysis. We aimed to prepare sterically stabilized trypsin-loaded liposomes (SSLT) and characterize their structure and fibrinolytic efficiency. Hydrogenated soybean phosphatidylcholine-based SSLT were prepared and their structure was studied by transmission electron microscopy combined with freeze fracture (FF-TEM), Fourier transform infrared spectroscopy (FT-IR), and small-angle X-ray scattering (SAXS). Fibrinolytic activity was examined at 45, 37, or 24(omicron)C on fibrin or plasma clots with turbidimetric and permeation-driven lysis assays. Trypsin was shown to be attached to the inner surface of vesicles (SAXS and FF-TEM) close to the lipid hydrophilic/hydrophobic interface (FT-IR). The thermosensitivity of SSLT was evidenced by enhanced fibrinolysis at 45(omicron)C: time to reduce the maximal turbidity to 20% decreased by 8.6% compared to 37(omicron)C and fibrin degradation product concentration in the permeation lysis assay was 2-fold to 5-fold higher than that at 24(omicron)C. SSLT exerted its fibrinolytic action on fibrin clots under both static and dynamic conditions, whereas plasma clot dissolution was observed only in the permeation-driven assay. The improved fibrinolytic efficiency of SSLT under dynamic conditions suggests that they may serve as a novel therapeutic candidate for dissolution of intravascular thrombi, which are typically exposed to permeation forces.