Role of surface charge in the internalization mechanisms of carboxy-methyl dextra

表面电荷在羧甲基右旋糖内化机制中的作用

• 批准号: 7777661
• 负责人: Magda M Latorre-Esteves
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF PUERTO RICO MAYAGUEZ
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7777661
• 关键词: Accounting; Active Biological Transport; Affect; Biochemical Pathway; Biocompatible; Biosensing Techniques; Caco-2 Cells; Cancer cell line; Carboxylic Acids; Cell Line; Cell Separation; Cells; Charge; Chemicals; Chemistry; Clathrin; Clinical; Colon Carcinoma; Contrast Media; Coupled; Cytosol; DNA delivery; Development; Dextrans; Diagnostic; Discrimination; Drug Delivery Systems; Drug Design; Electrostatics; Endocytosis; Epithelial; Fever; Human; Image; In Vitro; Kinetics; Laser Scanning Confocal Microscopy; Link; Liquid substance; Lysosomes; MCF7 cell; Magnetic Resonance Imaging; Magnetism; Malignant Neoplasms; Mass Spectrum Analysis; Medical; Medicine; Membrane Microdomains; Methods; Molecular; Molecular Weight; Pathway interactions; Pattern; Pharmaceutical Preparations; Phase; Plasma; Polymers; Property; Research; Research Personnel; Role; Shapes; Sorting - Cell Movement; Surface; System; Time; Tumor Cell Line; Work; Wound Healing; cancer therapy; carboxymethyl dextran; cell type; chemical property; clinical application; design; dextran; functional group; hydrophilicity; hyperthermia treatment; improved; magnetite ferrosoferric oxide; malignant breast neoplasm; nanoparticle; neoplastic cell; passive transport; public health relevance; receptor; receptor mediated endocytosis; tool; vector

DESCRIPTION (provided by applicant): Fundamental understanding of the role of nanoparticle physicochemical properties (such as size, charge, hydrophilicity, etc.) on internalization into cells and intracellular localization is a requirement for the rational design and optimization of nanoparticles for imaging, sensing, drug or DNA delivery, cell sorting, and many other clinical applications. The long term objective of this application is to elucidate the relationship between nanoparticle physicochemical properties and internalization and fate of nanoparticles in cells. As a first specific aim, methods will be refined to use established pharmacological agents to systematically inhibit molecular internalization pathways of cells in vitro and demonstrate that such methods can be applied to elucidate the mechanism of internalization of carboxy-methyl dextran coated magnetite nanoparticles into the cancer cell lines of epithelial origin Caco-2 (human colon cancer derived) and MFC-7 (human breast cancer derived). The methods to be refined will permit discrimination between active and passive transport mechanisms, clathrin- dependent pathways (and the sub-divisions of receptor mediated endocytosis and fluid phase endocytosis), and clathrin-independent pathways (and the subdivisions of caveolar and lipid raft-dependent endocytosis). As a second specific aim, the role of surface charge on internalization and intracellular localization of carboxy- methyl dextran coated nanoparticles will be assessed by using magnetite nanoparticles of similar size, coated with carboxy-methyl dextran of the same molecular weight but varying degrees of carboxylic acid substitution. Nanoparticle internalization through each of the above-mentioned molecular pathways will be assessed qualitatively using confocal laser scanning microscopy and quantitatively using inductively coupled plasma mass spectrometry. The methods to be refined through the proposed work should find applicability in systematic study of nanoparticle internalization mechanisms for a wide array of inorganic core/organic shell nanoparticles and a wide array of target cells. The specific information to be obtained on mechanisms and effect of surface charge in internalization of carboxy-methyl dextran coated nanoparticles will be applicable to the rational design of magnetic nanoparticles with potentially improved efficacy as MRI contrast agents, magnetically targeted drug delivery vectors, and in magnetic fluid hyperthermia treatment of cancer. PUBLIC HEALTH RELEVANCE: The relationship between nanoparticle physicochemical properties and internalization in cells is a key step in developing medical applications of magnetic nanoparticles, such as MRI contrast agents, sensing, drug delivery, and magnetic fluid hyperthermia cancer treatment. The proposed work will systematically study internalization mechanisms of magnetic nanoparticles in cells and determine the effect of surface charge on rate of internalization and intracellular localization of nanoparticles. The methods refined in this work will also impact study and development of other types of polymer coated inorganic core nanoparticles for medical applications.

描述（由申请人提供）：对纳米颗粒物理化学性质（如尺寸、电荷、亲水性等）的作用的基本理解对于用于成像、传感、药物或DNA递送、细胞分选和许多其他临床应用的纳米颗粒的合理设计和优化，需要对内化到细胞中和细胞内定位进行研究。本申请的长期目标是阐明纳米颗粒物理化学性质与细胞中纳米颗粒的内化和命运之间的关系。作为第一个具体目标，将改进方法以使用已建立的药理学试剂在体外系统地抑制细胞的分子内化途径，并证明这些方法可用于阐明羧甲基葡聚糖涂覆的磁铁矿纳米颗粒内化到上皮来源的癌细胞系Caco-2（人结肠癌来源）和MFC-7（人乳腺癌来源）中的机制。待完善的方法将允许区分主动和被动转运机制、网格蛋白依赖性途径（以及受体介导的内吞作用和液相内吞作用的细分）和网格蛋白非依赖性途径（以及小窝和脂筏依赖性内吞作用的细分）。作为第二个具体目的，表面电荷对羧甲基葡聚糖包被的纳米颗粒的内化和细胞内定位的作用将通过使用类似尺寸的磁铁矿纳米颗粒来评估，所述磁铁矿纳米颗粒包被有相同分子量但不同程度的羧酸取代的羧甲基葡聚糖。将使用共聚焦激光扫描显微镜定性评估通过上述分子途径中的每一种的纳米颗粒内化，并使用电感耦合等离子体质谱法定量评估纳米颗粒内化。通过所提出的工作进行改进的方法应该在系统研究纳米颗粒内化机制的广泛阵列的无机核/有机壳纳米颗粒和广泛阵列的靶细胞中找到适用性。获得的具体信息的机制和影响的表面电荷的内在化的羧甲基葡聚糖涂层的纳米粒子将适用于合理设计的磁性纳米粒子具有潜在的改善疗效的MRI造影剂，磁靶向药物输送载体，并在磁流体热疗治疗癌症。 公共卫生相关性：纳米粒子的物理化学性质与细胞内化之间的关系是开发磁性纳米粒子的医学应用的关键步骤，例如MRI造影剂、传感、药物递送和磁流体热疗癌症治疗。本论文将系统地研究磁性纳米颗粒在细胞内的内化机制，并确定表面电荷对纳米颗粒内化速率和细胞内定位的影响。在这项工作中改进的方法也将影响其他类型的聚合物包覆的无机核纳米粒子的研究和开发，用于医疗应用。

项目成果

Effect of surface charge on the colloidal stability and in vitro uptake of carboxymethyl dextran-coated iron oxide nanoparticles.

• DOI: 10.1007/s11051-013-1874-0
• 发表时间: 2013-08-01
• 期刊: JOURNAL OF NANOPARTICLE RESEARCH
• 影响因子: 2.5
• 作者: Ayala, Vanessa;Herrera, Adriana P.;Latorre-Esteves, Magda;Torres-Lugo, Madeline;Rinaldi, Carlos
• 通讯作者: Rinaldi, Carlos

Synthesis, Stability, Cellular Uptake, and Blood Circulation Time of Carboxymethyl-Inulin Coated Magnetic Nanoparticles.

• DOI: 10.1039/c3tb20256a
• 发表时间: 2013-06-14
• 期刊: Journal of materials chemistry. B
• 作者: Santiago-Rodríguez L;Lafontaine MM;Castro C;Méndez-Vega J;Latorre-Esteves M;Juan EJ;Mora E;Torres-Lugo M;Rinaldi C
• 通讯作者: Rinaldi C