BACKGROUND & AIMS: :Macrophage-derived foam cells are a major component of atherosclerotic plaques and have an important role in the progression of atherosclerotic plaques, thus posing a great threat to human health. Photodynamic therapy (PDT) has emerged as a therapeutic strategy for atherosclerosis. Here, we investigated the effect of PDT mediated by upconversion fluorescent nanoparticles encapsulating chlorin e6 (UCNPs-Ce6) on the cholesterol efflux of THP-1 macrophage-derived foam cells and explored the possible mechanism of this effect. First, we found that PDT notably enhanced the cholesterol efflux and the induction of autophagy in both THP-1 and peritoneal macrophage-derived foam cells. The autophagy inhibitor 3-methyladenine and an ATG5 siRNA significantly attenuated PDT-induced autophagy, which subsequently suppressed the ABCA1-mediated cholesterol efflux. Furthermore, the reactive oxygen species (ROS) produced by PDT were responsible for the induction of autophagy, which could be blocked by the ROS inhibitor N-acetyl cysteine (NAC). NAC also reversed the PDT-induced suppression of p-mTOR and p-Akt. Therefore, our findings demonstrate that PDT promotes cholesterol efflux by inducing autophagy, and the autophagy was mediated in part through the ROS/PI3K/Akt/mTOR signaling pathway in THP-1 and peritoneal macrophage-derived foam cells.

背景与目标： : 巨噬细胞衍生的泡沫细胞是动脉粥样硬化斑块的主要成分，在动脉粥样硬化斑块的进展中具有重要作用，因此对人类健康构成了巨大威胁。光动力疗法 (PDT) 已成为动脉粥样硬化的治疗策略。在这里，我们研究了封装了氯蛋白e6 (UCNPs-Ce6) 的上转换荧光纳米颗粒介导的PDT对THP-1巨噬细胞衍生泡沫细胞胆固醇流出的影响，并探索了这种作用的可能机制。首先，我们发现PDT显着增强了THP-1和腹膜巨噬细胞衍生的泡沫细胞中的胆固醇流出和自噬的诱导。自噬抑制剂3-甲基腺嘌呤和ATG5 siRNA显著减弱了PDT诱导的自噬，随后抑制了ABCA1-mediated胆固醇的流出。此外，PDT产生的活性氧 (ROS) 负责自噬的诱导，这可能被ROS抑制剂N-乙酰半胱氨酸 (NAC) 阻断。NAC还逆转了PDT诱导的p-mTOR和p-Akt的抑制。因此，我们的发现表明，PDT通过诱导自噬促进胆固醇流出，自噬部分通过ROS/PI3K/Akt/mTOR信号通路介导THP-1和腹膜巨噬细胞衍生的泡沫细胞。

BACKGROUND & AIMS: :Non-invasive gene monitoring is important for most gene therapy applications to ensure selective gene transfer to specific cells or tissues. We developed a non-invasive imaging system to assess the location and persistence of gene expression by anchoring an anti-dansyl (DNS) single-chain antibody (DNS receptor) on the cell surface to trap DNS-derivatized imaging probes. DNS hapten was covalently attached to cross-linked iron oxide (CLIO) to form a 39+/-0.5 nm DNS-CLIO nanoparticle imaging probe. DNS-CLIO specifically bound to DNS receptors but not to a control single-chain antibody receptor. DNS-CLIO (100 microM Fe) was non-toxic to both B16/DNS (DNS receptor positive) and B16/phOx (control receptor positive) cells. Magnetic resonance (MR) imaging could detect as few as 10% B16/DNS cells in a mixture in vitro. Importantly, DNS-CLIO specifically bound to a B16/DNS tumor, which markedly reduced signal intensity. Similar results were also shown with DNS quantum dots, which specifically targeted CT26/DNS cells but not control CT26/phOx cells both in vitro and in vivo. These results demonstrate that DNS nanoparticles can systemically monitor the expression of DNS receptor in vivo by feasible imaging systems. This targeting strategy may provide a valuable tool to estimate the efficacy and specificity of different gene delivery systems and optimize gene therapy protocols in the clinic.

背景与目标： : 非侵入性基因监测对于大多数基因治疗应用是重要的，以确保选择性基因转移到特定的细胞或组织。我们开发了一种非侵入性成像系统，通过将抗dansyl (DNS) 单链抗体 (DNS受体) 锚定在细胞表面以捕获DNS衍生的成像探针来评估基因表达的位置和持久性。DNS半抗原与交联的氧化铁 (CLIO) 共价连接以形成39 +/-0.5纳米DNS-CLIO纳米颗粒成像探针。DNS-CLIO与DNS受体特异性结合，但不与对照单链抗体受体结合。DNS-CLIO (100微微Fe) 对B16/DNS (DNS受体阳性) 和B16/phOx (对照受体阳性) 细胞均无毒。磁共振 (MR) 成像可以在体外检测到混合物中的10% 个B16/DNS细胞。重要的是，DNS-CLIO与B16/DNS肿瘤特异性结合，从而显着降低了信号强度。DNS量子点也显示了类似的结果，该量子点专门针对CT26/DNS细胞，但在体外和体内均不控制CT26/phOx细胞。这些结果表明，DNS纳米颗粒可以通过可行的成像系统系统地监测体内DNS受体的表达。这种靶向策略可能为评估不同基因递送系统的功效和特异性以及优化临床中的基因治疗方案提供有价值的工具。

BACKGROUND & AIMS: :The three-component nanoparticle of this investigation consisted of an anti-type I regulatory subunit alpha of the cyclic AMP-dependent protein kinase A (RIalpha) antisense phosphorodiamidate morpholino (MORF) oligomer, a tat peptide and the anti-HER2 Herceptin antibody each biotinylated and each linked via streptavidin and tested in SUM190 (HER2+), SUM149 (HER2-) and SK-BR-3 (HER2+) cells in culture, using both radioactivity and fluorescent labels on the antisense and control sense MORF. Within the nanoparticle, the antibody provides specific binding to the target cells, the tat improves cellular delivery and the MORF provides the specific retention of the radioactivity in the target cell nucleus. The results show that within the nanoparticle, the Herceptin was still able to bind to its determinant; that the MORF escaped entrapment with its mRNA-binding ability preserved and that the tat maintained its carrier function. Fluorescence microscopy showed evidence of antisense MORF internalization, separation from Herceptin and migration to the nucleus. In conclusion, streptavidin appears to provide an easy means of mixing and matching components to improve the tumor-specific targeting, cell membrane transport, pharmacokinetics and other properties of antisense and other oligomers. Combining the three components of this investigation with streptavidin apparently did not interfere with the properties of each component in cell culture and significantly improved delivery.

背景与目标： : 本研究的三组分纳米颗粒由环AMP依赖性蛋白激酶A (RIalpha) 反义磷酸二胺酯吗啉代 (MORF) 寡聚物的抗I型调节亚基 α 组成，tat肽和anti-HER2的赫赛汀抗体各自生物素化，并分别通过链霉亲和素连接，并在培养的SUM190 (HER2 +) 、SUM149 (HER2-) 和SK-BR-3 (HER2 +) 细胞中进行测试，使用反义和对照意义MORF上的放射性和荧光标记。在纳米颗粒中，抗体提供与靶细胞的特异性结合，tat改善细胞递送，而MORF提供放射性在靶细胞细胞核中的特异性保留。结果表明，在纳米颗粒中，赫赛汀仍然能够结合其决定簇; MORF逃脱了截留，保留了其mRNA结合能力，tat保持了其载体功能。荧光显微镜显示反义MORF内在化，从赫赛汀分离并迁移到细胞核的证据。总之，链霉亲和素似乎提供了一种混合和匹配组分的简便方法，以改善反义和其他寡聚物的肿瘤特异性靶向，细胞膜转运，药代动力学和其他特性。将本研究的三个成分与链霉亲和素结合使用显然不会干扰细胞培养中每个成分的特性，并且显着改善了递送。

BACKGROUND & AIMS: :Current standard of care for many CNS tumors involves surgical resection followed by chemotherapy and/or radiation. Some pediatric brain tumor types are infiltrative and diffuse in nature, which reduces the role for surgery. Furthermore, children are extremely vulnerable to neurological sequelae from surgery and radiation therapy, thus alternative approaches are in critical need. As molecular targets underlying various cancers become more clearly defined, there is an increasing push for targeted gene therapies. Viral vectors and nonviral nanoparticles have been thoroughly investigated for gene delivery and show promise as vectors for gene therapy for pediatric brain cancer. Here, we review inorganic and organic materials in development for nanoparticle gene delivery to the brain with a particular focus on safety.

背景与目标： : 目前许多中枢神经系统肿瘤的治疗标准包括手术切除，然后进行化疗和/或放疗。一些小儿脑肿瘤类型本质上是浸润性和弥漫性的，这降低了手术的作用。此外，儿童极易受到手术和放射治疗的神经系统后遗症的影响，因此迫切需要替代方法。随着各种癌症潜在的分子靶标变得更加明确，靶向基因治疗的推动力越来越大。已经对病毒载体和非病毒纳米颗粒进行了彻底的基因传递研究，并显示出有望作为小儿脑癌基因治疗的载体。在这里，我们回顾了纳米颗粒基因传递到大脑的开发中的无机和有机材料，特别关注安全性。

BACKGROUND & AIMS: :Nanoparticle albumin-bound (nab)-technology is an industrial applicable manufacturing method for the preparation of albumin-based drug carriers of poorly water-soluble drugs. In the present study the advantages of nanotechnology, albumin as an endogenous protein with the capability of high tumor enrichment and the selective light activation of the photosensitizer Temoporfin (mTHPC) were combined to a new delivery system for oncological use. The herewith provided well-established photodynamic therapy may enable a beneficial alternative for the treatment of solid tumors. In the present study a reproducible method for the preparation of stable mTHPC-albumin nanoparticles via nab-technology was established. The nanoparticles were physicochemically characterized with regard to particle size and size distribution and the impact of this preparation method on nanoparticle as well as mTHPC stability was investigated. Nanoparticles with improved colloidal stability over a broad pH range and in the presence of physiological NaCl concentrations were achieved in high yield. Due to high pressure homogenization a certain oxidative decay of mTHPC was observed. Cell culture experiments revealed an effective cellular uptake of mTHPC in a cholangiocarcinoma cell line (TFK-1). After light-activation high cytotoxicity was shown for photosensitizer loaded nanoparticles enabling the application of the proposed formulation in photodynamic therapy.

背景与目标： : 纳米颗粒白蛋白结合 (nab)-技术是一种工业上适用的制造方法，用于制备水溶性差的药物的白蛋白基药物载体。在本研究中，纳米技术的优势，白蛋白作为一种内源性蛋白，具有高肿瘤富集能力和光敏剂temooporfin (mTHPC) 的选择性光激活，被结合为一种新的肿瘤递送系统。由此提供的完善的光动力疗法可以为实体瘤的治疗提供有益的替代方法。在本研究中，建立了一种通过nab技术制备稳定的mTHPC-白蛋白纳米颗粒的可重复方法。纳米颗粒在粒径和粒径分布方面进行了物理化学表征，并研究了该制备方法对纳米颗粒以及mTHPC稳定性的影响。在较宽的pH范围内以及在生理NaCl浓度存在下，具有改善的胶体稳定性的纳米颗粒以高收率获得。由于高压均质化，观察到mTHPC的某些氧化衰减。细胞培养实验揭示了在胆管癌细胞系 (TFK-1) 中有效摄取mTHPC的细胞。光激活后，对负载有光敏剂的纳米颗粒显示出高细胞毒性，使所提出的制剂能够在光动力疗法中应用。

BACKGROUND & AIMS: BACKGROUND:Capsaicin (CPS) is a highly selective agonist of the transient receptor potential vanilloid type 1 (TRPV1) with a nanomolar affinity. High doses or prolonged exposure to CPS induces TRPV1 defunctionalization and, although this effect is currently used for the treatment of thermal hyperalgesia in chronic pain conditions, it is responsible of detrimental effects, such as denervation of sensory fibers. The aim of the present study was to formulate CPS loaded lipid nanocarriers (CPS-LN) in order to optimize CPS release, thus preventing TRPV1 internalization and degradation. METHODS:CPS-LNs were formulated and characterized by in vitro studies. The activation of TRPV1 receptors after CPS-LN administration was evaluated by measuring spontaneous pain that was induced by local injection into the plantar surface of the mouse hind-paw. Moreover, the expression of TRPV1 in the skin was evaluated by western blot analysis in CPS-LN injected mice and then compared to a standard CPS solution (CPS-STD). RESULTS:CPS inclusion in LN induced a lower pain response when compared to CPS-STD; further, it prevented TRPV1 down-regulation in the skin, while CPS-STD induced a significant reduction of TRPV1 expression. CONCLUSIONS:Drug encapsulation in lipid nanoparticles produced an optimization of CPS release, thus reducing mice pain behavior and avoiding the effects that are caused by TRPV1 defunctionalization related to a prolonged activation of this receptor.

背景与目标：

BACKGROUND & AIMS: :One of the major challenges for new therapeutics molecules to enter the clinic remains improving their bioavailability and cellular uptake. Therefore, delivery has become a key stone in therapeutic development and several technologies have been designed to improve cellular uptake of therapeutic molecules, including cell-penetrating peptides (CPPs) or protein transduction domain (PTD). PTDs or CPPs were discovered twenty years ago, based on the potency of several proteins to enter cells and nowadays, numerous peptide carriers have been described and successfully applied for ex vivo and in vivo delivery of varying therapeutic molecules. Two CPP-strategies have been reported; the first one requires chemical linkage between the drug and the carrier for cellular drug internalization and the second is based on the formation of stable complexes with drugs depending on their chemical nature. Peptide-Based-Nanoparticle Devices (PBND), correspond to short amphipathic peptides able to form stable nanoparticles with proteins and/or nucleic acids. Three PBND-families, PEP, MPG and CADY have been described, these carriers mainly enter cells independently of the endosomal pathway and efficiently deliver cargoes in a large variety of challenging cell lines as well as in animal models. This review will focus on the structure/function relationship of the PBND: CADY, PEP and MPG, in the general context of drug delivery. It will also highlight the requirement of primary or secondary amphipathic carriers for in vitro and in vivo delivery of therapeutic molecules and provide an update of their pre-clinical evaluation.

背景与目标： : 新的治疗分子进入临床的主要挑战之一仍然是提高其生物利用度和细胞摄取。因此，递送已成为治疗发展的关键石，并且已经设计了几种技术来改善治疗分子的细胞摄取，包括细胞穿透肽 (CPPs) 或蛋白质转导结构域 (PTD)。PTDs或cpp是在20年前发现的，基于几种蛋白质进入细胞的能力，如今，已经描述了许多肽载体，并成功地用于各种治疗分子的体外和体内递送。已经报道了两种CPP策略; 第一种需要药物与载体之间的化学连接以进行细胞药物内在化，第二种是基于药物的化学性质与药物形成稳定的复合物。基于肽的纳米颗粒装置 (PBND) 对应于能够与蛋白质和/或核酸形成稳定纳米颗粒的短两亲肽。已经描述了三个PBND家族，PEP，MPG和CADY，这些载体主要独立于内体途径进入细胞，并在各种具有挑战性的细胞系以及动物模型中有效地输送货物。这篇综述将集中在药物递送的一般背景下，PBND: CADY，PEP和MPG的结构/功能关系。它还将强调一级或二级两亲性载体在体外和体内递送治疗分子的需求，并提供其临床前评估的更新。

BACKGROUND & AIMS: :We studied the effectiveness of trilysine (Lys3), tetralysine (Lys4), pentalysine (Lys5), and poly-l-lysine (PLL) (MW 50000) on lambda-DNA nanoparticle formation and characterized the size, shape, and stability of nanoparticles. Light scattering experiments showed EC50 (lysine concentration at 50% DNA compaction) values of approximately 0.0036, 2, and 20 micromol/L, respectively, for PLL, Lys5, and Lys4 at 10 mM [Na+]. Plots of log EC50 versus log [Na+] showed positive slopes of 1.09 and 1.7, respectively, for Lys4 and Lys5 and a negative slope of -0.1 for PLL. Hydrodynamic radii of oligolysine condensed particles increased (48-173 nm) with increasing [Na+], whereas no significant change occurred to nanoparticles formed with PLL. There was an increase in the size of nanoparticles formed with Lys5 at >40 degrees C, whereas no such change occurred with PLL. The DNA melting temperature increased with oligolysine concentration. These results indicate distinct differences in the mechanism(s) by which oligolysines and PLL provoke DNA condensation to nanoparticles.

背景与目标： : 我们研究了三赖氨酸 (Lys3)，四赖氨酸 (Lys4)，五赖氨酸 (Lys5) 和聚-l-赖氨酸 (PLL) (MW 50000) 对lambda-DNA纳米颗粒形成的有效性，并表征了纳米颗粒的大小，形状和稳定性。光散射实验显示，对于PLL，Lys5和Lys4在10 mM [Na +] 下的EC50 (50% DNA压实时的赖氨酸浓度) 值分别约为0.0036、2和20微摩尔/L。log EC50与log [Na] 的图分别显示Lys4和Lys5的正斜率为1.09和1.7，PLL的负斜率为-0.1。低聚赖氨酸缩合颗粒的流体力学半径随着 [Na] 的增加而增加 (48-173 nm)，而PLL形成的纳米颗粒没有显着变化。在> 40摄氏度下，用Lys5形成的纳米颗粒的尺寸增加，而PLL没有发生这种变化。DNA熔化温度随低聚赖氨酸浓度的增加而增加。这些结果表明低聚物和PLL引发DNA缩合到纳米颗粒的机理存在明显差异。

BACKGROUND & AIMS: :Cerium oxide nanoparticles (CNPs) as an antioxidant have been used frequently to attenuate hyperglycaemia oxidative damage in different organs. We investigated the impact CNPs on the qualitative and quantitative sperm parameters, spermatogenesis and NFE2-related factor 2 (Nrf2) expression as a major contributor of antioxidant defence in the male diabetic rats. Twenty-four male rats were divided into four groups. Controls received only mouse food and water. Second group were treated with CNPs (30 mg kg-1  day-1 ) for 2 weeks. Rats in third group received streptozotocin (STZ) (60 mg/kg). In fourth group, animals became diabetic and received CNPs (30 mg kg-1  day-1 ) for 2 weeks. The results showed a significant abnormality in the sperm parameters and histopathological patterns of testes in the diabetic group compared to the control group and CNPs treatment significantly improved all testicular parameters. Following CNPs administration, sperm DNA fragmentation significantly reduced in the STZ-treated rats. Moreover, after CNPs intake in the STZ-treated rats, Nfr2 expression levels increased significantly. Overall, CNPs administration on the diabetic rates can attenuate detrimental effects of diabetes on the sperm potential fertility, sperm parameters, DNA integrity and Nrf2 expression levels. This study gives a future prospect to determine the role of CNPs in the context of diabetes.

背景与目标： 氧化铈纳米颗粒 (CNPs) 作为一种抗氧化剂，经常用于减轻不同器官的高血糖氧化损伤。我们研究了CNPs对定性和定量精子参数，精子发生和NFE2-related因子2 (Nrf2) 表达的影响，这是雄性糖尿病大鼠抗氧化防御的主要贡献者。将24只雄性大鼠分为四组。对照组只收到老鼠的食物和水。第二组用CNPs (30 mg kg-1天-1) 治疗2周。第三组大鼠接受链脲佐菌素 (STZ) (60 mg/kg)。在第四组中，动物成为糖尿病患者并接受CNPs (30 mg kg-1天-1) 治疗2周。结果显示，与对照组相比，糖尿病组的精子参数和睾丸的组织病理学模式明显异常，CNPs治疗显着改善了所有睾丸参数。施用CNPs后，STZ处理的大鼠的精子DNA断裂显着减少。此外，在STZ处理的大鼠中摄入CNPs后，Nfr2表达水平显着增加。总体而言，CNPs给药对糖尿病的影响可以减轻糖尿病对精子的潜在生育力，精子参数，DNA完整性和Nrf2表达水平的不利影响。这项研究为确定CNPs在糖尿病中的作用提供了未来的前景。

BACKGROUND & AIMS: :Despite great advances in the fight against cancer, traditional chemotherapy has been hindered by the dose dependent adverse side effects that reduce the usable doses for effective therapy. This has been associated to drug resistance in tumor cells that often cause relapse and therapy failure. These drawbacks have been tackled by combining different therapeutic regiments that prevent drug resistance while decreasing the chemotherapy dose required for efficacious ablation of cancer. In fact, new metallic compounds have been in a continuous development to extend the existing chemotherapy arsenal for these combined regimens. Here, we demonstrate that combination of a metallic compound (TS265), previously characterized by our group, with photothermy circumvents cells resistant to Doxorubicin (DOX). We first engendered a colorectal carcinoma cell line (HCT116) highly resistant to DOX, whose viability was diminished after administration of TS265. Cancer cell death was potentiated by challenging these cells with 14 nm spherical gold nanoparticles followed by laser irradiation at 532 nm. The combination of TS265 with photothermy lead to 65% cell death of the DOX resistant cells without impacting healthy cells. These results support the use of combined chemotherapy and photothermy in the visible spectrum as an efficient tool for drug resistant tumors.

背景与目标： : 尽管在对抗癌症方面取得了巨大进步，但传统的化学疗法受到剂量依赖性不良副作用的阻碍，这些副作用减少了有效治疗的可用剂量。这与经常导致复发和治疗失败的肿瘤细胞的耐药性有关。通过结合不同的治疗方案来解决这些缺点，这些方案可防止耐药性，同时减少有效消融癌症所需的化学疗法剂量。实际上，新的金属化合物一直在不断发展，以扩展这些联合疗法的现有化疗武器库。在这里，我们证明了先前由我们小组表征的金属化合物 (TS265) 与光热的结合，可以避免对阿霉素 (DOX) 具有抗性的细胞。我们首先产生了对DOX具有高度抗性的大肠癌细胞系 (HCT116)，该细胞系在施用ts265后活力降低。通过用14 nm球形金纳米颗粒挑战这些细胞，然后在532  nm处进行激光照射，可以增强癌细胞的死亡。TS265与光热的组合导致DOX抗性细胞的65% 细胞死亡，而不影响健康细胞。这些结果支持在可见光谱中使用联合化疗和照相疗法作为耐药肿瘤的有效工具。

BACKGROUND & AIMS: :Abundant desmoplastic stroma, which typically exists in pancreatic ductal adenocarcinoma (PDAC), can act as a natural protective physical barrier rendering insufficient drug delivery and penetration. To address this issue, we herein report a two-step sequential delivery strategy for enhanced pancreatic cancer therapy. In this sequential strategy, the nitric oxide (NO) donor S-nitroso-N-acetylpenicillamine (SNAP) loaded liposomes (Lip-SNAP) were firstly delivered to pancreatic stellate cells (PSCs) in tumor tissue to inhibit the production of dense stroma, by inhibiting the expression of TGF-β1 and its downstream profibrotic signal transduction. Therefore, the PSC-mediated desmoplastic reaction could be suppressed by inhibiting the expression of fibronectin, α-SMA and collagen. The gemcitabine (GEM) loaded liposomes (Lip-GEM) were administrated subsequently. The enhanced intratumoral penetration of Lip-GEM was then achieved due to the stromal disruption in consequence of NO treatment, thus significantly improving the drug delivery efficiency. The tumor growth inhibition of the two-step sequential delivery of Lip-SNAP and Lip-GEM was investigated on both subcutaneous and orthotopic tumor mouse models, to show the remarkably improved therapeutic efficacy of GEM. Such NO-induced stromal depletion provides a general strategy to overcome the blockage of desmoplastic stroma on other therapeutic agents.

背景与目标： : 通常存在于胰腺导管腺癌 (PDAC) 中的丰富的促结缔组织间质可以作为天然的保护性物理屏障，从而导致药物输送和渗透不足。为了解决这个问题，我们在此报告了用于增强胰腺癌治疗的两步顺序递送策略。在这种顺序策略中，首先将一氧化氮 (NO) 供体S-亚硝基-N-乙酰青霉胺 (SNAP) 脂质体 (Lip-SNAP) 递送到肿瘤组织中的胰腺星状细胞 (psc)，以抑制致密基质的产生，通过抑制TGF-β1的表达及其下游纤维化前信号转导。因此，可以通过抑制纤连蛋白，α-SMA和胶原蛋白的表达来抑制PSC介导的促纤维增生反应。随后给药吉西他滨 (GEM) 负载的脂质体 (Lip-GEM)。随后，由于不进行治疗而导致基质破坏，从而增强了Lip-GEM的瘤内渗透，从而显着提高了药物输送效率。在皮下和原位肿瘤小鼠模型上研究了Lip-SNAP和Lip-GEM两步顺序递送的肿瘤生长抑制作用，以显示GEM的治疗效果显着提高。这种NO诱导的基质耗竭提供了一种通用策略，可以克服其他治疗剂对促增生性基质的阻塞。

BACKGROUND & AIMS: :We have previously introduced the use of the biomaterial chitosan to form chitosan/siRNA nanoparticles for gene silencing protocols. This present study shows that the physicochemical properties (size, zeta potential, morphology and complex stability) and in vitro gene silencing of chitosan/siRNA nanoparticles are strongly dependent on chitosan molecular weight (Mw) and degree of deacetylation (DD). High Mw and DD chitosan resulted in the formation of discrete stable nanoparticles approximately 200 nm in size. Chitosan/siRNA formulations (N:P 50) prepared with low Mw (approximately 10 kDa) showed almost no knockdown of endogenous enhanced green fluorescent protein (EGFP) in H1299 human lung carcinoma cells, whereas those prepared from higher Mw (64.8-170 kDa) and DD (approximately 80%) showed greater gene silencing ranging between 45% and 65%. The highest gene silencing efficiency (80%) was achieved using chitosan/siRNA nanoparticles at N:P 150 using higher Mw (114 and 170 kDa) and DD (84%) that correlated with formation of stable nanoparticles of approximately 200 nm. In conclusion, this work confirms the application of chitosan as a non-viral carrier for siRNA and the importance of polymeric properties for the optimisation of gene silencing using chitosan/siRNA nanoparticles.

背景与目标： : 我们之前已经介绍了使用生物材料壳聚糖形成壳聚糖/siRNA纳米颗粒的基因沉默方案。本研究表明，壳聚糖/siRNA纳米颗粒的理化性质 (大小，zeta电位，形态和复合物稳定性) 和体外基因沉默强烈依赖于壳聚糖分子量 (Mw) 和脱乙酰度 (DD)。高Mw和DD壳聚糖导致形成尺寸约200 nm的离散稳定纳米颗粒。低Mw (约10 kDa) 制备的壳聚糖/siRNA制剂 (N:P 50) 在H1299人肺癌细胞中几乎没有内源性增强绿色荧光蛋白 (EGFP) 的敲除，而由较高Mw (64.8-170 kDa) 和DD (约80%) 制备的那些显示出45% 和65% 之间更大的基因沉默。使用与形成约200纳米的稳定纳米颗粒相关的较高Mw (114和170 kDa) 和DD (84%)，在N:P 150使用壳聚糖/siRNA纳米颗粒获得最高的基因沉默效率 (80%)。总之，这项工作证实了壳聚糖作为siRNA的非病毒载体的应用，以及聚合物特性对于使用壳聚糖/siRNA纳米颗粒优化基因沉默的重要性。

BACKGROUND & AIMS: :The rapid, specific and sensitive detection of nucleic acids is of utmost importance for the identification of infectious agents, diagnosis and treatment of genetic diseases, and the detection of pathogens related to human health and safety. Here we report the development of a simple and sensitive nucleic acid sequence-based and Ru(bpy)3 (2+)-doped silica nanoparticle-labeled lateral flow assay which achieves low limit of detection by using fluorescencent nanoparticles. The detection of the synthetic nucleic acid sequences representative of Trypanosoma mRNA, the causative agent for African sleeping sickness, was utilized to demonstrate this assay. The 30 nm spherical Ru(bpy)3 (2+)-doped silica nanoparticles were prepared in aqueous medium by a novel method recently reported. The nanoparticles were modified by 3-glycidoxypropyl trimethoxysilane in order to conjugate to amine-capped oligonucleotide reporter probes. The fluorescent intensities of the fluorescent assays were quantified on a mictrotiter plate reader using a custom holder. The experimental results showed that the lateral flow fluorescent assay developed was more sensitive compared with the traditional colloidal gold test strips. The limit of detection for the fluorescent lateral flow assay developed is approximately 0.066 fmols as compared to approximately 15 fmols for the colloidal gold. The limit of detection can further be reduced about one order of magnitude when "dipstick" format was used.

背景与目标： : 核酸的快速，特异性和灵敏检测对于鉴定感染原，遗传疾病的诊断和治疗以及与人类健康和安全有关的病原体的检测至关重要。在这里，我们报告了一种简单而灵敏的基于核酸序列和Ru(bpy)3 (2) 掺杂的二氧化硅纳米颗粒标记的侧向流动测定法的开发，该测定法通过使用荧光纳米颗粒实现了低检测限。利用代表锥虫mRNA (非洲昏睡病的病原体) 的合成核酸序列的检测来证明该测定。通过最近报道的一种新方法，在水性介质中制备了30 nm球形Ru(bpy)3 (2) 掺杂的二氧化硅纳米颗粒。纳米颗粒被3-缩水甘油氧基丙基三甲氧基硅烷修饰，以缀合到胺封端的寡核苷酸报告探针。使用定制支架在mictrotiter板读取器上定量荧光测定的荧光强度。实验结果表明，与传统的胶体金试纸相比，开发的侧向流荧光测定法更灵敏。所开发的荧光侧向流测定法的检出限为约0.066 fmols，而胶体金的检出限为约15 fmols。当使用 “量油尺” 格式时，检测极限可以进一步降低约一个数量级。

BACKGROUND & AIMS: :Silver nanoparticles (AgNPS) are an important model system for studying potential environmental risks posed by the use of nanomaterials. So far there is no consensus as to whether toxicity is due to AgNPs themselves or Ag(+) ions leaching from their surfaces. In sea urchin Paracentrotus lividus, AgNPs cause dose dependent developmental defects such as delayed development, bodily asymmetry and shortened or irregular arms, as well as behavioural changes, particularly in swimming patterns, at concentration ∼0.3 mg/L AgNPs. It has been observed that AgNPs are more toxic than their equivalent Ag(+) ion dose.

背景与目标： : 银纳米颗粒 (AgNPS) 是研究纳米材料使用带来的潜在环境风险的重要模型系统。到目前为止，关于毒性是由于AgNPs本身还是Ag () 离子从其表面浸出所致，尚无共识。在海胆副牛鱼中，AgNPs会导致剂量依赖性发育缺陷，例如发育延迟、身体不对称和手臂缩短或不规则，以及行为变化，特别是在游泳模式中，浓度为0.3毫克/升AgNPs。已经观察到AgNPs比其等效的Ag () 离子剂量更具毒性。

BACKGROUND & AIMS: :The increasing use of nano-sized materials in our environment, and in many consumer products, dictates new safety concerns. In particular, adequate experimental models are needed to evaluate skin toxicity of metal oxide ions, commonly found in cosmetic and dermatologic preparations. We have addressed the biological effects of topically applied copper oxide (CuO) nanoparticles in human skin organ cultures, using light and electron microscopy, and biochemical tests. Nanoparticles were more toxic than micro-sized particles, and their effects were stronger when supplied in growth medium than in topical application. Still topically applied CuO nanoparticles induced inflammatory cytokine secretion and necrosis, especially in epidermis deprived of its protective cornea. Since nanoparticle penetration was not seen, we propose that they may adhere to skin surface, react with the local acidic environment, and generate soluble ions that make their way to inner sites. This work illustrates the abilities of skin organ culture to evaluate the biological effects of topically-applied materials on skin in vitro.

背景与目标： : 纳米材料在我们的环境和许多消费产品中越来越多地使用，这引起了新的安全问题。特别是，需要足够的实验模型来评估金属氧化物离子的皮肤毒性，通常在化妆品和皮肤病学制剂中发现。我们已经使用光学和电子显微镜以及生化测试解决了局部应用的氧化铜 (CuO) 纳米颗粒在人体皮肤器官培养物中的生物学作用。纳米颗粒比微型颗粒更具毒性，并且在生长培养基中提供时，其作用比在局部应用中更强。仍局部应用的CuO纳米颗粒可诱导炎性细胞因子分泌和坏死，尤其是在缺乏保护性角膜的表皮中。由于未看到纳米颗粒的渗透，我们建议它们可能粘附在皮肤表面，与局部酸性环境反应，并产生可溶离子，从而进入内部位置。这项工作说明了皮肤器官培养物评估体外局部应用材料对皮肤的生物学作用的能力。