装载吲哚美辛肝素类纳米胶束的制备及抗凝作用
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摘要
肝素(hep)作为一种抗凝剂,在临床医学上广泛应用。本文以肝素为亲水段、去氧胆酸(DOCA)为疏水段合成了一种肝素类两亲性的聚合物(hep-DOCA,HD),并通过水相自组装方法,制备纳米胶束(HD-IDM),装载具有抗血栓作用的吲哚美辛(IDM),协同实现材料的抗凝血功能。通过动态光散射、Zeta电势和透射电子显微镜(TEM)等技术手段表征了纳米胶束结构和性能。当纳米胶束载药量为0. 0913 mg/mL时,载药纳米胶束浓度为0. 4 g/L,此时细胞存活率为92. 81%,溶血率为0. 83%,表明载药胶束具有良好的生物相容性。肝素钠,hep-DOCA和HD-IDM的全血凝血指数分别为86. 48%、77. 47%和89. 53%,全血凝血时间分别为927、837和965 s,血栓凝块实验中产生的血栓质量分别为0. 11、0. 20和0. 07 g,证明载药纳米胶束具有优良的抗凝血效果。
Heparin has been used as anti-thrombotic agents clinically. In this work,the copolymer based on hydrophilic heparin and hydrophobic deoxycholic acid( hep-DOCA, HD) was synthesized. Indometacin( IDM)-loaded HD micelles were prepared through aqueous self-assembly to enhance the anti-thrombotic capability of micelles synergistically and characterized by dynamic light scattering, Zeta potential and transmission electron microscopry( TEM). When the drug content of the nanomicelles was 0. 0913 g/L,the concentration of the drug-loaded nanomicelle was 0. 4 g/L,and the cell survival rate and hemolysis rate were 92. 81% and 0. 83%,indicating that the drug-loaded nanomicelles had good biocompatibility. The whole blood coagulation indexes of heparin sodium, hep-DOCA and HD-IDM were 86. 48%,77. 47% and 89. 53%,respectively. The whole blood coagulation time was 927,837 and 965 s,respectively. The thrombus mass produced in the thrombus clotting experiment was 0. 11,0. 20 and 0. 07 g,respectively,which confirmed that the drug-loaded nanomicelles had excellent anticoagulant effect.
Heparin has been used as anti-thrombotic agents clinically. In this work,the copolymer based on hydrophilic heparin and hydrophobic deoxycholic acid( hep-DOCA, HD) was synthesized. Indometacin( IDM)-loaded HD micelles were prepared through aqueous self-assembly to enhance the anti-thrombotic capability of micelles synergistically and characterized by dynamic light scattering, Zeta potential and transmission electron microscopry( TEM). When the drug content of the nanomicelles was 0. 0913 g/L,the concentration of the drug-loaded nanomicelle was 0. 4 g/L,and the cell survival rate and hemolysis rate were 92. 81% and 0. 83%,indicating that the drug-loaded nanomicelles had good biocompatibility. The whole blood coagulation indexes of heparin sodium, hep-DOCA and HD-IDM were 86. 48%,77. 47% and 89. 53%,respectively. The whole blood coagulation time was 927,837 and 965 s,respectively. The thrombus mass produced in the thrombus clotting experiment was 0. 11,0. 20 and 0. 07 g,respectively,which confirmed that the drug-loaded nanomicelles had excellent anticoagulant effect.
引文
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[12]Lana G,Lua B,Wang T,et al. Chitosan/Gelatin Composite Sponge is an Absorbable Surgical Hemostatic Agent[J].Colloid Surf B,2015,136:1026-1034.
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[14]Kuang H,Wang Y,Hu J,et al. A Method for Preparation of an Internal Layer of Artificial Vascular Graft Co-Modified with Salvianolic Acid B and Heparin[J]. ACS Appl Mater Interfaces,2018,10(23):19365-19372.
[2]SHI Qiang,LUAN Shifang,JIN Jing,et al. Hemocompatibility of Commodity Polymers Modified with Chemical and Biological Method[J]. Mater China,2014,33(4):212-223(in Chinese).石强,栾世方,金晶,等.通用高分子材料的化学和生物改性及其血液相容性研究[J].中国材料进展,2014,33(4):212-223.
[3]Feng S,Hu Y,Peng S,et al. Nanoparticles Responsive to the Inflammatory Microenvironment for Targeted Treatment of Arterial Restenosis[J]. Biomaterials,2016,105:167-184.
[4]Kawata H,Yoshiko Uesugi Y,Soeda H,et al. A New Drug Delivery System for Intravenous Coronary Thrombolysis with Thrombus Targeting and Stealth Activity Recoverable by Ultrasound[J]. J Am Coll Cardiol,2012,60(24):2550-2557.
[5]Wen Z,Jie J,Zhang Y,et al. A Self-assembled Polyjuglanin Nanoparticle Loaded with Doxorubicin and Anti-Kras siRNA for Attenuating Multidrug Resistance in Human Lung Cancer[J]. Biochem Biophys Res Commun,2017,493(4):1430-1437.
[6]Lianga Y,Kiick K. Heparin-Functionalized Polymeric Biomaterials in Tissue Engineering and Drug Delivery Applications[J]. Acta Biomater,2014,10(4):1588-1600.
[7]Lee Y,Nam J,Shin H,et al. Conjugation of Low-Molecular-Weight Heparin and Deoxycholic Acid for the Development of a New Oral Anticoagulant Agent[J]. Circulation,2001,104(25):3116-3120.
[8]Park K,Kim K,Kwon I,et al. Preparation and Characterization of Self-assembled Nanoparticles of Heparin-Deoxycholic Acid Conjugates[J]. Langmuir,2004,20(26):11726-11731.
[9]Khatun Z,Nurunnabi M,Lee D,et al. Optical Imaging,Biodistribution and Toxicity of Orally Administered Quantum Dots Loaded Heparin-Deoxycholic Acid[J]. Macromol Res,2015,23(7):686-695.
[10]Shearer J,Coker S,Carswell H,et al. Detrimental Effects of 2-Arachidonoylglycerol on Whole Blood Platelet Aggregation and on Cerebral Blood Flow after a Focal Ischemic Insult in Rats[J]. Am J Physiol-Heart C,2018,314(5):H967-H977.
[11]Lee Y,Nam J,Shin H,et al. Conjugation of Low-Molecular-Weight Heparin and Deoxycholic Acid for the Development of a New Oral Anticoagulant Agent[J]. Circulation,2001,104(25):3116-3120.
[12]Lana G,Lua B,Wang T,et al. Chitosan/Gelatin Composite Sponge is an Absorbable Surgical Hemostatic Agent[J].Colloid Surf B,2015,136:1026-1034.
[13]Hu M,Chang H,Zhang H,et al. Mechanical Adaptability of the MMP-Responsive Film Improves the Functionality of Endothelial Cell Monolayer[J]. Adv Healthcare Mater,2017,6(14):1601410.
[14]Kuang H,Wang Y,Hu J,et al. A Method for Preparation of an Internal Layer of Artificial Vascular Graft Co-Modified with Salvianolic Acid B and Heparin[J]. ACS Appl Mater Interfaces,2018,10(23):19365-19372.