聚乙二醇介导鹅掌楸悬浮细胞与CdSe／ZnS量子点纳米颗粒共孵育的互作特征

利用纳米材料介导的药物靶向治疗和动物细胞转基因等相关研究,日益受到人们的关注．但植物因存在细胞壁的障碍,无论原位还是离体细胞培养条件下,利用纳米技术进行基因转移均存在很大难度．因此设想,如通过纳米颗粒材料物理尺寸的改变和表面化学修饰,能改变纳米颗粒与植物细胞壁界面上的生物物理或生物化学特征,从而有利于纳米颗粒材料穿越植物细胞壁进入植物细胞,将对推动纳米技术在植物转基因领域中的应用产生重要意义．根据以上设想,研究了不同的共孵育时间和温度等条件下,杂交鹅掌楸的胚性悬浮细胞与经不同表面化学修饰的CdSe／ZnS纳米颗粒之间相互作用过程的细胞生物学特征,以及CdSe／ZnS量子点的细胞毒性．结果表明,在共孵育后3h以内,激光共聚焦显微镜和电子扫描显微镜下,均可观察到经表面后修饰带正电荷的CdSe／ZnS纳米颗粒．同时,胞吞进入细胞内部的表面携带正电荷的CdSe／ZnS纳米颗粒的量明显与共培养时间、温度有明显的依赖关系,表明它们可以通过细胞的液相胞吞作用进入杂交鹅掌楸细胞内,且不影响细胞的活性;而表面带负电荷的CdSe／ZnS纳米颗粒则主要聚集在细胞外壁附近．在培养溶液中添加20％（质量比）聚乙二醇,可进一步提高鹅掌楸细胞胞吞CdSe／ZnS纳米颗粒的量和减轻CdSe／ZnS纳米颗粒的细胞毒性．本研究表明,以表面携带正电荷的CdSe／ZnS量子点纳米材料作为基因载体,在植物悬浮细胞的转基因研究和应用中具有广泛的前景．     

纳米氧化锌对斑马鱼鳃组织的氧化损伤

正纳米颗粒是指粒径在1—100 nm,表面积较大,具有块状颗粒所没有的特有性质。由于纳米颗粒具有独特的理化性质,近年来合成和生产纳米颗粒的产量大大增加[1]。纳米氧化物颗粒在科研和工业产品的应用正在逐年增加,科学家预测依靠纳米技术创造的产品产生的价值,到2015年可能达到1万亿美元[2]。随着纳米颗粒的使用,它对人类健康和环境的风险也随之增加。因此了解纳米颗粒的不利影响,确定对生物的影响,确保纳米材料的安全使用是十分必要的[3]。     

综述——纳米材料在组织工程中的应用：磁性纳米材料在组织工程研究中的应用

磁性纳米材料因其独特的理化性质在组织工程研究中被广泛地应用．本文主要从磁性纳米材料的表面化学活性、磁学性质以及生物应用磁性纳米材料的主要合成方法等几方面,综述了近年来利用磁性纳米材料设计组织工程支架材料的相关研究进展,包括纳米条件下的生长因子及相关基因的包裹和释放、机械力学刺激、干细胞追踪以及细胞图案化．     

原子力显微镜在测定颗粒与细胞相互作用中的应用

凭借独特的尺寸效应和理化性质,纳微颗粒在生物医药领域的应用日益广泛,其与细胞的相互作用也备受关注,对其进行定量测量和机制研究愈发重要。目前,原子力显微镜(atomic force microscopy,AFM)由于具有高灵敏度(皮牛级)、高分辨率(纳米级)以及在生理环境中可进行实时检测等优势成为检测颗粒与细胞相互作用的重要工具。利用AFM检测颗粒与细胞相互作用,有助于确定作用过程中的重要参数,解释颗粒在药物递送、免疫响应和细胞力学等应用方面深层次的机制。本文中,笔者就原子力显微镜检测颗粒与细胞相互作用及其应用进行系统的综述,并对其未来的发展方向进行展望。     

纳米颗粒对细胞膜的作用

纳米颗粒在生物医学领域有着广泛的应用前景。在纳米颗粒与细胞相互作用的研究中,颗粒对细胞膜作用的相关研究,对揭示纳米颗粒的生物效应是至关重要的。纳米颗粒对细胞膜的影响有很多种,主要体现在对细胞膜结构和性质,以及对膜上生物大分子(蛋白质等)功能的影响等方面。这里综述了近年来纳米颗粒对细胞膜作用的相关研究成果,分别从颗粒的自身物理化学性质(尺寸、形状、表面形貌、亲疏水性质、表面电荷、特异性修饰等)、颗粒与细胞作用的环境因素,以及外界能量对颗粒与细胞膜作用的调控三个方面出发,就纳米颗粒作用对细胞膜影响的问题分别进行了分析和总结。     

生物基分子介导纳米金属材料的制备及其应用

纳米金属材料具有纳米晶强化效应、光吸收率大、较高的表面能和单磁畴性能等优点,因其在医药、化学催化、抗菌抑毒等方面发挥着越来越重要的作用而受到人们广泛关注。近年来,随着全球石化资源消耗与日俱增,环境污染加剧,基于可再生资源的生物基分子介导纳米材料的制备研究方兴未艾。生物基分子是指直接或间接来源于生物质的小分子或大分子物质,它们多数具有生物相容性好、低毒、可降解、来源广泛、价格低廉等优点。且由于生物基分子多数具有独特的理化性质,如具有生理活性的旋光性、酸碱两性、亲水亲油性以及易与金属离子络合等,其介导合成的纳米材料还兼具其独特功能性,比如消炎、抗癌、抗氧化、抗病毒以及降血糖血脂等,进一步拓宽了纳米金属材料的应用领域。文中对近年来基于生物基分子介导纳米金属材料的制备及应用进行全面综述,为开展相关研究提供参考。     

ABCA1的胞内运输及功能研究新进展

三磷酸腺苷结合盒转运体A1(ABCA1)具有介导细胞内脂质流出,维持细胞脂质稳态的功能．新生的ABCA1必须经过胞内运输和各种化学修饰等过程,最终成为具有功能的成熟转运体,才能行使其转运脂质的功能,因此,ABCA1在胞内的运输过程和正确质膜定位对其介导胆固醇流出的功能至关重要．目前ABCA1相关研究主要集中于脂质转运方面,并提出各种胆固醇流出机制的模型,如通道转运模型、蘑菇状突起模型和胞吞-胞吐转运模型等．最近研究显示,ABCA1还具有调节质膜脂筏结构、参与免疫和炎症调节等新功能．本文主要针对ABCA1的胞内运输过程以及各种功能做一综述,以期为动脉粥样硬化相关疾病提供新的治疗靶点和途径．     

纳米材料在污染水体和土壤修复中的应用

纳米材料由于其特有的理化性质,如强的吸附性能、高的催化效率,不仅克服了传统污染水体和污染土壤修复技术的不足,而且表现出更高的修复效率。因此,利用纳米材料对污染环境进行修复已成为当今环境领域的研究热点。本文对纳米Fe0、表面修饰的纳米材料、纳米螯合剂等纳米材料近几年在污染水体和污染土壤修复中的进展进行了归纳和总结,并且对其在环境科学与工程领域应用的研究重点进行了展望。     

纳米材料生物效应研究进展

随着纳米技术的快速发展,纳米材料在医学成像、疾病诊断、药物传输、癌症治疗、基因治疗等领域的应用和基础研究也在飞速发展.同时,纳米材料的这些有益应用使得人体通过吸入、经口、皮肤吸收和静脉注射等不同方式受到暴露.当纳米材料与生物体系发生相互作用时,有可能产生负面生物学效应,而这些潜在的毒理效应都是未知的.综述了纳米材料在生物医学领域巨大的应用前景,关注其对心血管系统、呼吸系统及转运到其他器官可能造成的负面效应,并探讨了纳米颗粒在引起心血管疾病及肺部炎症方面的可能机理与作用途径.最后对纳米材料的安全性评估和研究重点进行了总结.     

微波合成靶向核仁的荧光碳纳米颗粒研究

核仁是细胞内重要的亚核结构,其在恶性病的演变过程中扮演重要角色,是病理学家诊断癌症的重要指标.尽管核仁如此关键,但到目前为止,核仁的荧光探针寥寥无几.本文以水杨酸和1,8-二氨基萘为反应物,通过微波消解法合成了一种新型荧光碳纳米颗粒(FCNs),采用透射电子显微镜、动态光散射仪、傅里叶红外光谱仪、紫外分光光度计、荧光光谱仪等对其物理、化学、光学性质进行了表征、分析.借助激光扫描共聚焦等技术对FCNs的细胞摄取机制及分布进行了探究.实验结果表明,所合成碳纳米颗粒尺寸均匀,最佳激发波长在348 nm,对应的最大发射峰为432 nm,荧光量子产率为17.8%,荧光寿命为1.13 ns,其表面含有丰富的氨基和羟基,光稳定性强且毒性极低,可实现对细胞核仁染色,并且随着共孵育时间的延长,进入细胞的量越多,靶向核仁更明显.此外,经过对FCNs细胞摄取路径的考察,发现FCNs是通过小窝介导的路径被內吞.该研究为碳基纳米材料在亚细胞器靶向成像的应用方面提供了有力的工具和新思路.     

Targeting and cellular trafficking of magnetic nanoparticles for prostate cancer imaging

Antibody-conjugated iron oxide nanoparticles offer a specific and sensitive tool to enhance magnetic resonance (MR) images of both local and metastatic cancer. Prostate-specific membrane antigen (PSMA) is predominantly expressed on the neovasculature of solid tumors and on the surface of prostate cells, with enhanced expression following androgen deprivation therapy. Biotinylated anti-PSMA antibody was conjugated to streptavidin-labeled iron oxide nanoparticles and used in MR imaging and confocal laser scanning microscopic imaging studies using LNCaP prostate cancer cells. Labeled iron oxide nanoparticles are internalized by receptor-mediated endocytosis, which involves the formation of clathrin-coated vesicles. Endocytosed particles are not targeted to the Golgi apparatus for recycling but instead accumulate within lysosomes. In T(1)-weighted MR images, the signal enhancement owing to the magnetic particles was greater for cells with magnetic particles bound to the cell surface than for cells that internalized the particles. However, the location of the particles (surface vs internal) did not significantly alter their effect on T(2)-weighted images. Our findings indicate that targeting prostate cancer cells using PSMA offers a specific and sensitive technique for enhancing MR images.     

TAT and HA2 Facilitate Cellular Uptake of Gold Nanoparticles but Do Not Lead to Cytosolic Localisation

The methods currently available to deliver functional labels and drugs to the cell cytosol are inefficient and this constitutes a major obstacle to cell biology (delivery of sensors and imaging probes) and therapy (drug access to the cell internal machinery). As cell membranes are impermeable to most molecular cargos, viral peptides have been used to bolster their internalisation through endocytosis and help their release to the cytosol by bursting the endosomal vesicles. However, conflicting results have been reported on the extent of the cytosolic delivery achieved. To evaluate their potential, we used gold nanoparticles as model cargos and systematically assessed how the functionalisation of their surface by either or both of the viral peptides TAT and HA2 influenced their intracellular delivery. We evaluated the number of gold nanoparticles present in cells after internalisation using photothermal microscopy and their subcellular localisation by electron microscopy. While their uptake increased when the TAT and/or HA2 viral peptides were present on their surface, we did not observe a significant cytosolic delivery of the gold nanoparticles.     

Surface immunoglobulin on rabbit lymphoid cells. II. Ultrastructural distribution of b4 allotypic determinants and morphology of spleen lymphocytes

Surface immunoglobulin allotypic determinants on rabbit spleen lymphoid cells are ultrastructurally localized by labeling with antiallotype antisera and soluble complexes of ferritin and rabbit antiferritin of a given allotype. At 0 °C surface Ig is visualized in patches on the membrane of 54% of the spleen lymphocytes examined. Four morphologically distinct categories of spleen lymphocytes display different amounts of labeled surface Ig. Type I cells are essentially identical to peripheral blood lymphocytes and demonstrate rapid endocytosis of surface Ig at 37 °C. Type II cells greater amounts of surface Ig, demonstrate little endocytosis, and are consistent with lymphoblast cells. Type III cells have the greatest amount of surface Ig, reveal some endocytosis, and are morphologically consistent with proplasmacytes and plasmablast cells. Type IV cells are immature plasma cells and have very little detectable surface Ig. The percentage of each cell type making up the labeled population is Type I, 28%; Type II, 21%; Type III, 50%, and Type IV, 1%. Immunoferritin labeled Ig determinants may be modulated from the surface of these cells at 37 °C by endocytosis and/or by shedding after reaction with anti-Ig antisera.     

Peptide ligand‐mediated endocytosis of nanoparticles to cancer cells: Cell receptor‐binding‐ versus cell membrane‐penetrating peptides

The endocytosis‐mediating performances of two types of peptide ligands, cell receptor binding peptide (CRBP) and cell membrane penetrating peptide (CMPP), were analyzed and compared using a common carrier of peptide ligands‐human ferritin heavy chain (hFTH) nanoparticle. Twenty‐four copies of a CMPP(human immunodeficiency virus‐derived TAT peptide) and/or a CRBP (peptide ligand with strong and specific affinity for either human integrin(α_vβ₃) or epidermal growth factor receptor I (EGFR) that is overexpressed on various cancer cells) were genetically presented on the surface of each hFTH nanopariticle. The quantitative level of endocytosis and intracellular localization of fluorescence dye‐labeled CRBP‐ and CMPP‐presenting nanoparticles were estimated in the in vitro cultures of integrin‐ and EGFR‐overexpressing cancer and human dermal fibroblast cells(control). From the cancer cell cultures treated with the CMPP‐ and CRBP‐presenting nanoparticles, it was notable that CRBPs resulted in quantitatively higher level of endocytosis than CMPP (TAT) and successfully transported the nanoparticles to the cytosol of cancer cells depending on concentration and treatment period of time, whereas TAT‐mediated endocytosis localized most of the nanoparticles within endosomal vesicles under the same conditions. These novel findings provide highly useful informations to many researchers both in academia and in industry who are interested in developing anticancer drug delivery systems/carriers.     

Redundant and Distinct Functions for Dynamin-1 and Dynamin-2 Isoforms

A role for dynamin in clathrin-mediated endocytosis is now well established. However, mammals express three closely related, tissue-specific dynamin isoforms, each with multiple splice variants. Thus, an important question is whether these isoforms and splice variants function in vesicle formation from distinct intracellular organelles. There are conflicting data as to a role for dynamin-2 in vesicle budding from the TGN. To resolve this issue, we compared the effects of overexpression of dominant-negative mutants of dynamin-1 (the neuronal isoform) and dynamin-2 (the ubiquitously expressed isoform) on endocytic and biosynthetic membrane trafficking in HeLa cells and polarized MDCK cells. Both dyn1(K44A) and dyn2(K44A) were potent inhibitors of receptor-mediated endocytosis; however neither mutant directly affected other membrane trafficking events, including transport mediated by four distinct classes of vesicles budding from the TGN. Dyn2(K44A) more potently inhibited receptor-mediated endocytosis than dyn1(K44A) in HeLa cells and at the basolateral surface of MDCK cells. In contrast, dyn1(K44A) more potently inhibited endocytosis at the apical surface of MDCK cells. The two dynamin isoforms have redundant functions in endocytic vesicle formation, but can be targeted to and function differentially at subdomains of the plasma membrane.     

Localized functionalization of individual colloidal carriers for cell targeting and imaging

Fabricating drug particles for therapeutic delivery and imaging presents important challenges in the design of the particle surfaces. Drug nanoparticle surfaces are currently functionalized with site-specific targeting ligands, biocompatible polymers, or fluorophore-polymer conjugates for specific imaging. However, if these functionalizations were to be synthesized on the drug carrier in localized, nanoscale regions on the particle surface, new schemes of drug delivery could be realized. Here we describe the use of our particle lithography technique that enables the synthesis of individual colloidal carrier assemblies that can be imaged and targeted to integrin-expressing cells. We show localized adhesion specificity for cells expressing the target integrin followed by receptor-mediated endocytosis. With the addition of localized delivery by adding drug nanoparticles to a specific region on the particle surface, our colloidal carrier assemblies have the potential to target, deliver therapeutic agents to, sense, and image diseased endothelium.     

Overviews on the cellular uptake mechanism of polysaccharide colloidal nanoparticles

Nanoparticulate drug/gene carriers have gained much attention in the past decades because of their versatile and tunable properties. However, efficacy of the therapeutic agents can be further enhanced using naturally occurring materials‐based nanoparticles. Polysaccharides are an emerging class of biopolymers; therefore, they are generally considered to be safe, non‐toxic, biocompatible and biodegradable. Considering that the target of nanoparticle‐based therapeutic strategies is localization of biomedical agents in subcellular compartments, a detailed understanding of the cellular mechanism involved in the uptake of polysaccharide‐based nanoparticles is essential for safe and efficient therapeutic applications. Uptake of the nanoparticles by the cellular systems occurs with a process known as endocytosis and is influenced by the physicochemical characteristics of nanoparticles such as size, shape and surface chemistry as well as the employed experimental conditions. In this study, we highlight the main endocytosis mechanisms responsible for the cellular uptake of polysaccharide nanoparticles containing drug/gene.     

Flow and shuttle of plasma membrane during endocytosis

A striking feature of endocytosis is the large amount of surface membrane that is brought into the cells through the formation of endocytic vesicles. Little is known about the fate of this membrane material. It is implausible that it would be destroyed in lysosomes, as the rate of turnover of the constituents of plasma membrane is much too low with respect to the rate of endocytosis in all cells studied so far. Conversely, plasma membrane fragments, internalized by endocytosis cannot merely be incorporated in lysosomes, as these organelles have been shown to maintain their size, despite continuous and active endocytosis. We present evidence that plasma membrane antigens, detected by means of specific antibodies, are internalized during endocytosis and reach lysosomes. They are thereafter returned back to cell surface. These results indicate the existence of a shuttle of membrane elements between the cell surface and lysosomes.     

Interactions of chitosan and its derivatives with cells (review)

Mechanisms of interaction between chitosan, various macromolecules or drug delivery systems and mammalian cells are reviewed. Modernly the role of different physicochemical properties of chitosan and chitosan nanoparticles on the mechanisms of cell bunding, endocytosis and redistribution are poorly understood.     

Atomic force microscopy-based cell nanostructure for ligand-conjugated quantum dot endocytosis

While it has been well demonstrated that quantum dots (QDs) play an important role inbiological labeling both in vitro and in vivo,there is no report describing the cellular nanostructure basis ofreceptor-mediated endocytosis.Here,nanostructure evolution responses to the endocytosis of transferrin(Tf)-conjugated QDs were characterized by atomic force microscopy (AFM).AFM-based nanostructureanalysis demonstrated that the Tf-conjugated QDs were specifically and tightly bound to the cell receptorsand the nanostructure evolution is highly correlated with the cell membrane receptor-mediated transduction.Consistently,confocal microscopic and flow cytometry results have demonstrated the specificity anddynamic property of Tf-QD binding and internalization.We found that the internalization of Tf-QD is linearlyrelated to time.Moreover,while the nanoparticles on the cell membrane increased,the endocytosis was stillvery active,suggesting that QD nanoparticles did not interfere sterically with the binding and function ofreceptors.Therefore,ligand-conjugated QDs are potentially useful in biological labeling of cells at a nanometerscale.