TALANTA
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2023
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IF 6.1
Xiujun Cao,Yuan Luo,Xueli Liu,Chunqing Shang,Jun Lu,Guoxin Song,Chunhui Deng
Hirudin, that is naturally occurring in leeches ( Hirudo medicinalis ) and known as the most potent natural inhibitor of thrombin, exerts double-edged effects in clinic application. It can be used as a therapeutic ingredient for cardiovascular disease, while it can be regarded as a toxic polypeptide with bleeding risk. Effective detection of hirudin in biological samples contributes greatly to reasonable therapy. In this study, we proposed a smart adsorbent based on affinity magnetic microspheres , where thrombin was immobilized for capturing hirudin in the animal serum. Aptamer was introduced as a ligand for linking the magnetic agarose microspheres and thrombin, thereby avoiding loss of biological activity of the enzyme to hirudin. Taken recombinant hirudin variant 2-Lys 47 (rHV2) as a model, we established a rapid and bio-specific extraction method coupled with liquid chromatography and quadrupole-time-of-flight mass spectrometry (LC-QTOF/MS) for determination of hirudin in the serum. Owing to this strategy, a low limit of detection (LOD) of rHV2 (0.5 nM), a good linearity with correlation coefficient of 0.9975, an acceptable precision with relative standard deviation (RSD) below 3.6% (n = 6) and acceptable recoveries ranging from 85.7% to 90.2% were achieved. Moreover, the functionalized magnetic composite could be reused for at least nine cycles. Our work combined the merits of affinity separation and advanced instrument analysis for hirudin, providing a new vision to precise determination of hirudin in medical and pharmaceutical fields.
研究领域:
生物化学
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药物分析
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医学检测
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DOI
PubMed
高影响因子
CHEMICAL RESEARCH IN CHINESE UNIVERSITIES
·
2023
·Vol. 39
·期 6
·1023-1030
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IF 3.1
Zheng Zhiwen,Dai Xin,Li Xueyang,Du Chang
M ore and more concerns about health bring the increasing demand for blood contact tissue engineering alternatives. In this paper, nanoparticles of poly(lactic-co-glycolic acid)/polyethyleneimine mixed with recombinant hirudin(rHNPs) were prepared by a double emulsion solvent volatilization method, which were then loaded onto the polycaprolactone(PCL) with polydopamine(PDA) coating to form the composite nanofibers of PCL/PDA/rHNPs. The hydrophilicity and mechanical properties of the composite nanofibers were improved significantly compared with pure PCL. The morphology kept almost unchanged after 30 d of degradation in phosphate buffer saline (PBS). The anticoagulant molecule of hirudin could be gradually released from the composite scaffolds through the degradation of rHNPs in vitro . When the concentration of rHNPs suspension was 5.0 mg/mL, the composite nanofibers could better promote the growth and proliferation of human umbilical vein endothelial cells(HUVECs). The anticoagulant ability of the composite nanofibers was also significantly improved in comparison with that of pure PCL. The design of controlled release anticoagulant materials would alleviate the sudden release of simple fixed hirudin, which could also provide a new idea for the development of novel blood contact materials.
研究领域:
生物材料学
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组织工程学
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药物释放系统
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DOI
高影响因子
Materials Science & Engineering C-Materials for Biological Applications
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2017
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IF 4.164
Qiongyu Wang,Fangfang Tu,Yunfei Liu,Yujin Zhang,Helei Li,Zhao Kang,Yin Yin,Jiannan Wang
Thrombus formation remains a particular challenge for small-diameter vascular grafts. In this study, the direct thrombin inhibitor hirudin (Hir) was used to modify silk fibroin films in an attempt to enhance its antithrombogenic properties. Hir was successfully attached to silk fibroin and uniformly distributed in the regenerative material. Hir-modified films showed good cytocompatibility, and supported adhesion and proliferation of fibroblasts (L929), human umbilical vascular endothelial cells (HUVECs) and human aortic smooth muscle cells (HASMCs). Proliferation of HAVSMCs was inhibited by increasing Hir concentration. Activated partial thrombin time (APTT), prothrombin time (PT) and thrombin time (TT) of Hir-modified silk fibroin tubular scaffolds (SFTSs) were all increased markedly compared with fresh rabbit blood, ethanol-treated SFTS and unmodified SFTS, demonstrating the improved antithrombogenicity of SFTSs following modification with Hir.
研究领域:
生物医学工程
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材料科学
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血液学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MEMBRANE SCIENCE
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2016
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IF 5.557
Jinglong Li,Fu Liu,Yan Qin,Jidong He,Zhu Xiong,Gang Deng,Qiang Li
Different from intensively used anticoagulant heparin, hirudin as a representative of natural bio-molecular inhibitors can form specific hirudin-thrombin complex and function as a serine proteinase to efficiently prevent blood coagulation with less bleeding adverse effect, no allergic reaction and immune of non-toxic reaction. We aim to develop a novel anticoagulant polylactide (PLA) membrane via immobilizing natural hirudin through the hydrogen bonding interaction. High amount of PVP (17.4 wt%) was first immobilized on the membrane surface through the interfacial crosslinking of P(VP-VTES). The abundant carbonyl groups with high inter-association constant (K=6000) on the membrane surface dominated the hydrogen bonding interaction with the carboxyl groups of hirudin [44] . The anti-clotting activity of PLA membrane increased with the active hirudin concentration. XPS, ATR-FTIR was conducted to confirm the chemistry evolution of PLA membranes immobilized by PVP and hirudin respectively. The intermediate PVP spacer promoted the hydrophilicity and effectively inhibited the platelet adhesion. While the enhanced hemocompatibility was distinctly revealed by the blood concretion four items (APTT, PT, TT and FIB) mainly thanks to the surface immobilization of hirudin. The anti-clotting stability of PLA membrane was evaluated with varied incubation time. The complement activation was also measured in terms of C3a and C5a. The anti-clotting PLA membrane showed excellent dialysis performances in terms of cleaning urea, creatinine, lysozyme while preserving BSA respectively.
研究领域:
生物材料学
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医学工程
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产品关联
DOI
高影响因子
Asian Journal of Pharmaceutical Sciences
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2022
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IF 9.273
Mengfang Wu,Tian Xia,Yaran Li,Tianfa Wang,Shijia Yang,Jinchao Yu,Qiaoyan Liang,Teng Shen,Min Yu,Bing Zhao
Cardiovascular disease is the leading cause of global mortality, with anticoagulant therapy being the main prevention and treatment strategy. Recombinant hirudin (r-hirudin) is a direct thrombin inhibitor that can potentially prevent thrombosis via subcutaneous (SC) and intravenous (IV) administration, but there is a risk of haemorrhage via SC and IV. Thus, microneedle (MN) provides painless and sanitary alternatives to syringes and oral administration. However, the current technological process for the micro mould is complicated and expensive. The micro mould obtained via three-dimensional (3D) printing is expected to save time and cost, as well as provide a diverse range of MNs. Therefore, we explored a method for MNs array model production based on 3D printing and translate it to micro mould that can be used for fabrication of dissolving MNs patch. The results show that r-hirudin-loaded and hyaluronic acid (HA)-based MNs can achieve transdermal drug delivery and exhibit significant potential in the prevention of thromboembolic disease without bleeding in animal models. These results indicate that based on 3D printing technology, MNs combined with r-hirudin are expected to achieve diverse customizable MNs and thus realize personalized transdermal anticoagulant delivery for minimally invasive and long-term treatment of thrombotic disease.
研究领域:
药剂学
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生物医学工程
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心血管疾病研究
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DOI
PubMed
高影响因子
Biomaterials Advances
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2023
·
IF 0
Zhiwen Zheng,Xin Dai,Xueyang Li,Chang Du
Blood-contacting materials with good mechanical property, excellent anticoagulant function and promoting effect on endothelialization are in great demand for clinical application such as vascular grafts in treating cardiovascular diseases. In this study, electrospinning nanofiber scaffolds of polycaprolactone (PCL) were functionalized by oxidative self-polymerization of dopamine (PDA) on the surface followed by the modification of anticoagulant recombinant hirudin (rH) molecules. The morphology, structure, mechanical property, degradation behavior, cellular compatibility and blood compatibility of the multifunctional PCL/PDA/rH nanofiber scaffolds were evaluated. The diameter of the nanofibers was between 270–1030 nm. The ultimate tensile strength of the scaffolds was around 4 MPa and the elastic modulus increased with the amount of rH. The degradation tests in vitro indicated that the nanofiber scaffolds began to crack on the 7th day, but still maintained the nanoscale architecture within a month. The cumulative release of rH from the nanofiber scaffold was up to 95.9 % at 30th day. The functionalized scaffolds promoted the adhesion and proliferation of endothelial cells, while resisting platelet adhesion and enhancing anticoagulation effects. The hemolysis ratios of all scaffolds were <2 %. The nanofiber scaffolds are promising candidates for vascular tissue engineering.
研究领域:
生物医学工程
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材料科学
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心血管医学
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产品关联
DOI
PubMed
Asian Journal of Pharmaceutical Sciences
·
2024
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IF 10.7
Mengying Cheng,Tianxiang Yue,Hong Wang,Lai Jiang,Qiaoling Huang,Fanzhu Li
Atherosclerosis (AS) is a progressive inflammatory disease, and thrombosis most probably leads to cardiovascular morbidity and mortality globally. Thrombolytic drugs alone cannot completely prevent thrombotic events, and treatments targeting thrombosis also need to regulate the inflammatory process. Based on the pathological process of AS dynamic development, biomimetic thrombus-targeted nanoparticles HMTL@PM were prepared. Hirudin and lumbrukinase, the effective substances of traditional Chinese medicine, were self-assembled under the action of tannic acid and Mn 2+ . HMTL@PM dissociated in the weakly acidic microenvironment of atherosclerosis and exhibited an excellent therapeutic effect of alleviating inflammation, dissolving thrombus, anticoagulation, and promoting cholesterol efflux. HMTL@PM effectively regulated the progression of AS and provided a new perspective for the development of drug delivery systems for AS therapy, which owned important research significance for reducing the mortality of cardiovascular and cerebrovascular diseases.
研究领域:
纳米技术
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药学
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心血管疾病
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治疗学
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DOI
PubMed
高影响因子
Small
·
2022
·Vol. 18
·期 51
·2203184
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IF 15.153
Shuting Li,Kai Zhang,Zhaoyu Ma,Weiyun Zhang,Zhiyong Song,Wenjing Wang,Heyou Han
Due to the high recurrence rate and mortality of venous thrombosis, there is an urgent need for research on antithrombotic strategies. Because of the short half-life, poor targeting capabilities, bleeding complications, and neurotoxic effects of conventional pharmacological thrombolysis methods, it is essential to develop an alternative strategy to noninvasive thrombolysis and decrease the recurrence rate of venous thrombosis. A platelet-mimetic porphyrin-based covalent organic framework-engineered melanin nanoplatform, to target delivery of hirudin to the vein thrombus site for noninvasive thrombolysis and effective anticoagulation, is first proposed. Owing to the thrombus-hosting properties of platelet membranes, the nanoplatform can target the thrombus site and then activate hyperthermia and reactive oxygen species for thrombolysis under near-infrared light irradiation. The photothermal therapy/photodynamic therapy combo can substantially improve the effectiveness (85.7%) of thrombolysis and prevent secondary embolism of larger fragments. Afterward, the highly loaded (97%) and slow-release hirudin (14 days) are effective in preventing the recurrence of blood clots without the danger of thrombocytopenia. The described biomimetic nanostructures offer a promising option for improving the efficacy of thrombolytic therapy and reducing the risk of bleeding complications in thrombus associated diseases.
研究领域:
纳米医学
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溶栓
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药理学
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生物医学工程
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材料科学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）