Au NPs/UCNPs复合纳米体系用于荧光成像引导下的肿瘤光热治疗的研究进展

近红外(NIR)光诱导的光热治疗(PTT)因其无创、非侵入、毒副作用低、可精准靶向治疗等特性,已成为肿瘤精准治疗的新型手段。凭借其独特的表面等离激元共振(SPR)特性及其高效的光热转换效率、生物毒性与良好的光稳定性,金纳米颗粒(Au NPs)已成为理想的光热治疗剂。而高质量成像技术是实现有效光热治疗的可靠有力的工具,尤其是多模态成像技术,比起单一成像方式具有更卓越的性能,为更全面、更精准的肿瘤成像提供了可能,显著提高了非侵入性医学治疗的潜力。NIR光激发的稀土上转换纳米颗粒(UCNPs),因其丰富的4f电子结构展现出磁性、荧光、X射线衰减和放射等多功能特性,使其作为造影剂在多模态成像领域展现了重要的应用前景。因此,构建NIR光诱导的Au NPs/UCNPs复合纳米体系,可用于多模态成像引导下的光热治疗,有望成为癌症诊疗的一种新策略。本文简单介绍了Au NPs、UCNPs的光学特性,重点综述了NIR光诱导的UCNPs-Au NPs(纳米壳、纳米棒、纳米团簇)复合纳米体系在癌症光热治疗领域的最新研究进展,并对其实现诊疗一体化的未来进行了展望。     

纳米金壳介导的光热效应对骨肉瘤干细胞体外杀伤作用的研究

目的:探究纳米金壳介导的光热效应对骨肉瘤干细胞的杀伤作用。方法:采用无血清悬浮培养法富集骨肉瘤干细胞,通过实时荧光定量PCR检测所富集细胞CD133、SOX2、NANOG、OCT4 mRNA的相对表达量以进行鉴定。将纳米金壳与骨肉瘤干细胞共培养,应用波长808 nm的近红外激光器(1.5 W/cm~2)进行照射以激发光热效应,分别用CCK8法检测光热疗法对骨肉瘤干细胞的增殖抑制率,Annexin C-Fitc/PI双染法检测光热疗法对骨肉瘤干细胞凋亡的影响。结果:成功富集了高表达CD133、SOX2、NANOG、OCT4 mRNA的骨肉瘤干细胞;纳米金壳介导的光热效应显著抑制了骨肉瘤干细胞的增殖率,当纳米金壳浓度在0～250μg/mL范围内时,增殖抑制率与浓度呈正相关;细胞凋亡结果显示,纳米金壳结合近红外照射组细胞凋亡率显著高于对照组。结论:纳米金壳介导的光热效应对骨肉瘤干细胞增殖有明显抑制作用,主要诱导骨肉瘤干细胞发生中晚期凋亡。     

半导体型单壁碳纳米管用于近红外二区(NIR-Ⅱ)深层组织光声成像

传统光声成像外源对比剂的光吸收主要集中在可见光区和传统近红外区(NIR,750～900 nm),开发具有更高光学组织穿透能力的近红外二区(NIR-Ⅱ,1 000～1 700 nm)光吸收外源对比剂对活体深层组织光声成像具有重要意义。本文中,作者选取了光吸收峰在1 000 nm左右的半导体型单壁碳纳米管为近红外二区光学吸收外源对比剂,测试了其在近红外二区激光激发下能够产生较强的光声效应。进一步地,作者通过将该纳米材料包埋在仿体组织的不同深度的位置,获得了仿体组织的深层光声成像,成像深度可达1.5 cm。试验结果表明,具有近红外二区光吸收能力的半导体型单壁碳纳米管在活体深层组织光声成像中有很大的应用潜力。     

核壳型磁性纳米粒子界面氨基的测定

目的:应用对硝基苯甲醛比色测定法测定磁性纳米粒子界面修饰的氨基.方法:合成核壳型磁性纳米粒子,并用AEAPS(氨乙基氨丙基聚二甲基硅氧烷)修饰其界面氨基,然后采用对硝基苯甲醛比色测定法测定不同反应条件下的磁性纳米粒子界面的氨基.结果:采用对硝基苯甲醛比色测定法可以测出在磁性纳米粒子界面连接的氨基量,不同反应条件下氨基连接量不同,氨基在磁性纳米粒子上的浓度为5.90至33.44 nmol/mg,最大相对量为1.63 umol/m2.结论:对硝基苯甲醛比色法不仅可以定量测定磁性纳米粒子界面氨基量,而且可以研究反应条件对磁性纳米粒子界面氨基的连接量的影响.     

靶向吲哚菁绿纳米探针介导的光声治疗

光声治疗是一种利用纳米材料的光声效应选择性破坏癌细胞的方法。本研究采用叶酸作为肿瘤靶向分子,以聚乙二醇包裹吲哚菁绿形成纳米粒子(ICG-PL-PEG-FA),利用此纳米粒子在近红外区的光吸收特性,开展光声治疗研究。实验结果表明,这种叶酸标记的纳米探针对高表达叶酸的EMT6细胞具有高靶向选择性和靶向光杀伤性。这种基于包含吲哚菁绿纳米探针的光声治疗将有潜力发展为一种安全,高效的癌症治疗技术。     

水解FeCl_3制备单分散P(St-co-AA)/Fe_2O_3亚微核壳粒子

用液相沉积法制备了壳层均匀、包裹致密的单分散P(St-co-AA)/Fe_2O_3亚微核壳粒子。用XRD、TEM和FESEM表征了该类粒子的物相、形貌及微观结构。结果表明用该法制备的核壳粒子,其壳层为Fe_2O_3晶粒,且均匀地包裹在乳胶粒子表面形成草莓状结构;改变FeCl_4溶液的用量和重复包裹次数能方便地调节P(St-co-AA)/Fe_2O_3亚微核壳粒子的壳层厚度。该核壳粒子可通过煅烧法来制备形状完整的单分散亚微中空磁球。     

巯基化壳聚糖包裹人参纳米颗粒的制备

目的:制备巯基化壳聚糖包裹的且具有良好的荧光敏感性的人参纳米颗粒.方法:采用分子嫁接法和溶剂挥发法,合成巯基化壳聚糖.利用得到的巯基化壳聚糖包裹人参纳米颗粒,制备巯基化壳聚糖人参纳米颗粒.加入对巯基敏感的荧光试剂,利用荧光显微镜对其荧光活性进行检测.结果:巯基化壳聚搪包裹的人参纳米颗粒形状规则,具有核-壳结构,湿态下平均粒径约为300nm;同时在生理pH下具有很强的荧光活性.结论:巯基化壳聚糖包裹的人参纳米颗粒具有良好的缓释作用和荧光敏感性.     

纳米金-聚乙烯亚胺基因载体的制备

目的:基因方法治疗癌症近年来取得了很大的突破,因此基因载体的构建显得尤为重要.其中纳米基因载体合成简单,成本低廉,并能够包裹、浓缩、保护核苷酸使其免受核酸酶降解,因此纳米材料广泛地应用于基因输送.我们拟开展聚乙烯亚胺-纳米金基因载体的制备及其表征.方法:采用层层包裹技术制备基因载体,首先通过柠檬酸钠还原法制备纳米金颗粒后,应用11-巯基十一烷酸对金颗粒进行修饰,使其表面带有羧基,然后进一步将带有氨基的低分子量聚乙烯亚胺与羧基进行连接.应用动态光散射(DLS),紫外可见光谱(UV)和透射电子显微镜(TEM)对构建的纳米基因载体进行表征.结果:成功制备了聚乙烯亚胺-纳米金基因载体,检测表明每一步制备出的产物纳米尺寸在20-30nm之间,液体均匀稳定,分散系数(PDI)在0.2以下,Zeta电位测定表明,每步的产物电荷变化与外层包裹的反应物有关.尽管金颗粒外层包裹聚乙烯亚胺,但是总体上纳米载体尺寸没有发生太大的变化,TEM检测表明每一步形成了均匀的、单分散的、球状的纳米颗粒.结论:我们通过层层包裹技术成功制备了聚乙烯亚胺-纳米金基因载体,在进一步开展的生物活性的检测中,希望通过纳米载体的携带作用,将基因转染进靶细胞,从而检测相关基因对靶细胞的沉默作用,提高基因药物的应用,为开发新型基因药物提供基础.     

基于L-半胱氨酸修饰的Fe3O4@TiO2复合纳米颗粒体外光动力疗法灭活HL60细胞的试验研究

本文采用共沉淀法制备了L-半胱氨酸(L-Cys)修饰的Fe3O4包裹TiO2(Fe3O4@TiO2/L-Cys)复合纳米粒子。通过透射电子显微镜(TEM),X射线衍射(XRD)和傅立叶变换红外光谱仪(FTIR)对复合纳米粒子进行了表征,并讨论了复合纳米粒子对HL60细胞体外光动力疗法(PDT)灭活的影响。并对其PDT灭活机制进行了初步探索。试验表明,Fe3O4@TiO2/L-Cys复合纳米粒子分散性高,生物相容性好,对细胞的暗毒性更低,并可以有效增强靶向性,提高PDT灭活效率,在410nm波长的光激发下,光照剂量为18J/cm^2的情况下,当TiO2与Fe3O4的比例为1∶3时,整体PDT效率最高。PDT灭活效率可达69.36%。     

pH和近红外光双响应的包裹二硫化钼纳米片和阿霉素的金属-有机框架ZIF-8用于肿瘤化学/光热协同治疗

利用金属-有机框架材料ZIF-8包裹二硫化钼(MoS_2)纳米片和阿霉素(DOX)构建一种可通过酸性pH和近红外(NIR)光双触发的肿瘤化学/光热协同治疗体系。首先,通过水热反应和超声处理制备粒径为～100 nm、厚度为0.3～1.4 nm的MoS_2纳米片。然后,通过一步法将可酸降解的金属-有机框架ZIF-8包裹在所制备的MoS_2纳米片上,并同时装载抗肿瘤药物DOX,形成装载DOX的ZIF-8包裹MoS_2纳米复合物(DOX/MoS_2@ZIF-8)。将该纳米复合物应用到肿瘤细胞的化学/光热协同治疗:当处于酸性条件(例如:溶酶体中pH大约为5)和NIR激光(780 nm,2.1 W/cm~2)照射的情况下,DOX/MoS_2@ZIF-8纳米复合物上包裹的ZIF-8金属-有机框架会发生酸降解,释放出所包裹的DOX,细胞质中的DOX可以进入细胞核中诱导细胞凋亡;同时,MoS_2纳米片能够将光能转换为热能,光致高温同样能诱导细胞凋亡,因此,化学/光热协同肿瘤治疗得以实现。细胞存活率试验证明:该DOX/MoS_2@ZIF-8纳米复合物在SMMC-7721细胞上表现出良好的化学/光热协同治疗作用,能够对肿瘤细胞进行高效地杀伤。     

Optimized Synthesis of PEG-Encapsulated Gold Nanorods for Improved Stability and Its Application in OCT Imaging with Enhanced Contrast

Gold nanorods (GNRs) are synthesized with a surfactant template, which often poses toxicity issues for biomedical applications. In addition, blue shift of longitudinal surface plasmon resonance (LSPR) peak of GNR is an inherent problem that needs to be addressed for time-course studies. In this work, we resolve these issues by optimizing the encapsulation of GNRs with polyethylene glycol (PEG) where biocompatibility is improved by ～20 % and blue shift over a period of 8 days is reduced from 20 nm in the case of CTAB-GNR to 2 nm for PEG-encapsulated GNR. The encapsulated GNRs were then bioconjugated for targeted dark-field imaging of cancer cells. As an application, we also demonstrate the contrast-enhancing capability of GNRs in optical coherence tomography (OCT) imaging of tumor xenograft where the LSPR closely matches the OCT excitation wavelength. Our study proves that incorporating GNRs enhances the contrast of tumor tissue interfaces along with a considerable broadening in OCT depth profile by six times.     

Evaluation of size,morphology, concentration,and surface effect of gold nanoparticles on X-ray attenuation in computed tomography

Increasing attention has been focused on the use of nanostructures as contrast enhancement agents in medical imaging, especially in computed tomography (CT). To date, gold nanoparticles (GNPs) have been demonstrated to have great potential as contrast agents for CT imaging. This study was designed to evaluate any effect on X-ray attenuation that might result from employing GNPs with a variety of shapes, sizes, surface chemistries, and concentrations. Gold nanorods (GNRs) and spherical GNPs were synthesized for this application. X-ray attenuation was quantified by Hounsfield unit (HU) in CT. Our findings indicated that smaller spherical GNPs (13 nm) had higher X-ray attenuation than larger ones (60 nm) and GNRs with larger aspect ratio exhibited great effect on X-ray attenuation. Moreover, poly ethylene glycol (PEG) coating on GNRs declined X-ray attenuation as a result of limiting the aggregation of GNRs. We observed X-ray attenuation increased when mass concentration of GNPs was elevated. Overall, smaller spherical GNPs can be suggested as a better alternative to Omnipaque, a good contrast agent for CT imaging. This data can be also considered for the application of gold nanostructures in radiation dose enhancement where nanoparticles with high X-ray attenuation are applied.     

Tissue Distribution and Efficacy of Gold Nanorods Coupled with Laser Induced Photoplasmonic Therapy in Ehrlich Carcinoma Solid Tumor Model

Gold nanorods (GNR) within tumor microregions are characterized by their ability to absorb near IR light and emit heat in what is called photoplasmonic effect. Yet, the efficacy of nanoparticles is limited due to intratumoral tissue distribution reasons. In addition, distribution of GNRs to normal tissue might result in non specific toxicity. In the current study, we are assessing the intratumoral and tissue distribution of PEGylated GNRs on the top of its antitumor characteristics when given intravenously or intratumoral to solid tumor bearing mice and coupled with laser photoplasmonic sessions. PEGylated GNRs with a longitudinal size of less than 100 nm were prepared with aspect ratio of 4.6 showing strong surface plasmon absorption at wavelength 800 nm. Pharmacokinetics of GNR after single I.V. administration (0.1 mg/kg) showed very short systemic circulating time (less than 3 h). On the other hand, tissue distribution of I.V. GNR (0.1 mg/kg) to normal animals showed preferential deposition in spleen tissue. Repeated administration of I.V. GNR resulted in preferential accumulation in both liver and spleen tissues. In addition, I.V. administration of GNR to Ehrlich carcinoma tumor bearing mice resulted in similar tissue distribution; tumor accumulation and anti-tumor effect compared to intratumoral administration. In conclusion, the concentration of GNR achieved within tumors microregions after I.V. administration was comparable to I.T. administration and sufficient to elicit tumoral growth arrest when coupled with laser-aided photoplasmonic treatment.     

Oleyl-chitosan nanoparticles based on a dual probe for optical/MR imaging in vivo

Oleic acid-conjugated chitosan (oleyl-chitosan) is a powerful platform for encapsulating oleic acid-decorated iron oxide nanoparticles (ION), resulting in a good magnetic resonance imaging (MRI) probe. Oleyl-chitosan could self-assemble into core-shell structures in aqueous solution and provide the effective core compartment for loading ION. ION-loaded oleyl-chitosan nanoparticles showed good enhanced MRI sensitivity in a MR scanner. Cy5.5 dye was accessed to the oleyl-chitosan conjugate for near-infrared (NIR) in vivo optical imaging. After intravenous injection of ION-loaded Cy5.5-conjugated oleyl-chitosan (ION-Cy5.5-oleyl-chitosan) nanoparticles in tumor-bearing mice, both NIRF and MR imaging showed the detectable signal intensity and enhancement in tumor tissues via enhanced permeability and retention (EPR) effect. Tumor accumulation of the nanoparticles was confirmed through ex vivo fluorescence images and Prussian blue staining images in tumor tissues. It is concluded that ION-Cy5.5-oleyl-chitosan nanoparticle is highly an effective imaging probe for detecting tumor in vivo.     

Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters

Magnetic and plasmonic properties combined in a single nanoparticle provide a synergy that is advantageous in a number of biomedical applications including contrast enhancement in novel magnetomotive imaging modalities, simultaneous capture and detection of circulating tumor cells (CTCs), and multimodal molecular imaging combined with photothermal therapy of cancer cells. These applications have stimulated significant interest in development of protocols for synthesis of magneto-plasmonic nanoparticles with optical absorbance in the near-infrared (NIR) region and a strong magnetic moment. Here, we present a novel protocol for synthesis of such hybrid nanoparticles that is based on an oil-in-water microemulsion method. The unique feature of the protocol described herein is synthesis of magneto-plasmonic nanoparticles of various sizes from primary blocks which also have magneto-plasmonic characteristics. This approach yields nanoparticles with a high density of magnetic and plasmonic functionalities which are uniformly distributed throughout the nanoparticle volume. The hybrid nanoparticles can be easily functionalized by attaching antibodies through the Fc moiety leaving the Fab portion that is responsible for antigen binding available for targeting.     

A Localized Surface Plasmon Resonance Biosensor Based on Integrated Controllable Au2S/AuAgS-Coated Gold Nanorods Composite

In this work, we have demonstrated that the exquisite optical properties based on localized surface plasmon resonance (LSPR) of Au₂S/AuAgS-coated gold nanorods (Au₂S/AuAgS-coated GNRs) can be utilized to develop a simple and sensitive biosensor, and goat anti-human IgG can be detected by the human IgG probe as low as 0.2 nM. Moreover, we introduce an integrated LSPR biosensor constructed by integrating Au₂S/AuAgS-coated GNRs immobilized on glass slide and isolated Au₂S/AuAgS-coated GNRs in the form of liquid. The detection of target binding was performed via direct spectral changes induced by changes of refractive index in the vicinity of individual particles. The integrated LSPR optical biosensor is label-free, cost-effective, and easy to fabricate and requires only a visible/near-infrared spectrometer for detection purposes. Additionally, the investigation on the mutual influence of the two types of nanorods in the integrated LSPR biosensor was performed. The results of separate experiments indicate that the nanorods in the form of isolate or in integrated exhibit a similar behavior.     

Magnetic and contrast properties of labeled platelets for magnetomotive optical coherence tomography

This article introduces a new functional imaging paradigm that uses optical coherence tomography (OCT) to detect rehydrated, lyophilized platelets (RL platelets) that are in the preclinical trial stage and contain superparamagnetic iron oxides (SPIOs) approved by the U.S. Food and Drug Administration. Platelets are highly functional blood cells that detect and adhere to sites of vascular endothelial damage by forming primary hemostatic plugs. By applying magnetic gradient forces, induced nanoscale displacements (magnetomotion) of the SPIO-RL platelets are detected as optical phase shifts in OCT. In this article, we characterize the iron content and magnetic properties of SPIO-RL platelets, construct a model to predict their magnetomotion in a tissue medium, and demonstrate OCT imaging in tissue phantoms and ex vivo pig arteries. Tissue phantoms containing SPIO-RL platelets exhibited >3 dB contrast/noise ratio at ≥1.5 × 10⁹ platelets/cm³. OCT imaging was performed on ex vivo porcine arteries after infusion of SPIO-RL platelets, and specific contrast was obtained on an artery that was surface-damaged (P < 10⁻⁶). This may enable new technologies for in vivo monitoring of the adherence of SPIO-RL platelets to sites of bleeding and vascular damage, which is broadly applicable for assessing trauma and cardiovascular diseases.     

Plasmonic Effects of Dual-Metal Nanoparticle Layers for High-Performance Quantum Dot Solar Cells

To improve quantum dot solar cell performance, it is crucial to make efficient use of the available incident sunlight to ensure that the absorption is maximized. The ability of metal nanoparticles to concentrate incident sunlight via plasmon resonance can enhance the overall absorption of photovoltaic cells due to the strong confinement that results from near-field coupling or far-field scattering plasmonic effects. Therefore, to simultaneously and synergistically utilize both plasmonic effects, the placement of different plasmonic nanostructures at the appropriate locations in the device structure is also critical. Here, we introduce two different plasmonic nanoparticles, Au and Ag, to a colloidal PbS quantum dot heterojunction at the top and bottom interface of the electrodes for further improvement of the absorption in the visible and near-infrared spectral regions. The Ag nanoparticles exhibit strong scattering whereas the Au nanoparticles exhibit an intense optical effect in the wavelength region where the absorption of light of the PbS quantum dot is strongest. It is found that these dual-plasmon layers provide significantly improved short-circuit current and power conversion efficiency without any form of trade-off in terms of the fill factor and open-circuit voltage, which may result from the indirect contact between the plasmonic nanoparticles and colloidal quantum dot films.

     

Near-infrared fluorescent nanoparticles as combined MR/optical imaging probes

A number of quantitative three-dimensional tomographic near-infrared fluorescence imaging techniques have recently been developed and combined with MR imaging to yield highly detailed anatomic and molecular information in living organisms (1, 2). Here we describe magnetic nanoparticle based MR contrast agents that have a near-infrared fluorescence (NIRF) that is activated by certain enzymes. The probes are prepared by conjugation of arginyl peptides to cross-linked iron oxide amine (amino-CLIO), either by a disulfide linkage or a thioether linker, followed by the attachment of the indocyanine dye Cy5.5. The NIRF of disulfide-linked conjugate was activated by DTT, while the NIRF of thioether-linked conjugate was activated by trypsin. Fluorescent quenching of the attached fluorochrome occurs in part due to the interaction with iron oxide, as evident by the activation of fluorescence with DTT when nanoparticles that have less than one dye attached per particle. With a SC injection of the probe, axillary and brachial lymph nodes were darkened on MR images and easily delineated by NIRF imaging. The probes may provide the basis for a new class of so-called smart nanoparticles, capable of pinpointing their position through their magnetic properties, while providing information on their environment by optical imaging techniques.     

Contrast‐enhanced optical coherence tomography for melanoma detection: An in vitro study

Optical coherence tomography (OCT), with a high‐spatial resolution (<10 microns), intermediate penetration depth (~1.5 mm) and volumetric imaging capability is a great candidate to be used as a diagnostic‐assistant modality in dermatology. At this time, the accuracy of OCT for melanoma detection is lower than anticipated. In this letter, we studied for the first time, the use of a novel contrast agent consist of ultra‐small nanoparticles conjugated to a melanoma biomarker to improve the accuracy of OCT for differentiation of melanoma cells from nonmelanoma cells, in vitro. We call this approach SMall nanoparticle Aggregation‐enhanced Radiomics of Tumor (SMART)‐OCT imaging. This initial proof of concept study is the first step toward the broad utilization of this method for high accuracy all types of tumor detection applications.