MR imaging of the her2/neu and 9.2.27 tumor antigens using immunospecific contrast agents

Molecular imaging of tumor antigens using immunospecific magnetic resonance (MR) contrast agents is a rapidly evolving field, which can potentially aid in early disease detection, monitoring of treatment efficacy, and drug development. In this study, we designed, synthetized, and tested in vitro two novel monocrystalline iron oxide nanoparticles (MION) conjugated to antibodies against the her2/neu tyrosine kinase receptor and the 9.2.27 proteoglycane sulfate. MION was synthetized by coprecipitation of iron II and iron III salts in 12-kD dextran solution; antibody coupling was performed by reductive amination. The relaxivity of the conjugates was 24.1-29.1 mM(-1) s(-1), with 1.8 to 2.1 antibody molecules per nanoparticle. A panel of cultured melanoma and mammary cell lines was used for testing. The cells were incubated with the particles at 16-32 microg Fe/ml in culture medium for 3 h at 37 degrees C, and investigated with immune fluorescence, transmission electron microscopy (TEM), MRI of cell suspensions in gelatine, and spectrophotometric iron determination. All receptor-positive cell lines, but not the controls, showed receptor-specific immune fluorescence, and strong changes in T(2) signal intensity at 1.5 T. The changes in 1/T(2) were between 1.5 and 4.6 s(-1) and correlated with the amount of cell-bound iron (R = 0.92). The relaxivity of cell-bound MION increased to 55.9 +/- 10.4 mM(-1) s(-1). TEM showed anti-9.2.27 conjugates binding to the plasma membrane, while the anti-her2/neu conjugates underwent receptor-mediated endocytosis. In conclusion, we obtained receptor-specific T(2) MR contrast with novel covalently bound, multivalent MION conjugates with anti-9.2.27 and anti-her2/neu to image tumor surface antigens. This concept can potentially be expanded to a large number of targets and to in vivo applications.     

Enhancing photothermal cancer therapy by clustering gold nanoparticles into spherical polymeric nanoconstructs

Gold nanoparticles (AuNPs) have been proposed as agents for enhancing photothermal therapy in cancer and cardiovascular diseases. Different geometrical configurations have been used, ranging from spheres to rods and more complex star shapes, to modulate optical and ablating properties. In this work, multiple, ultra-small 6 nm AuNPs are encapsulated into larger spherical polymeric nanoconstructs (SPNs), made out of a poly(lactic acid-co-glycol acid) (PLGA) core stabilized by a superficial lipid-PEG monolayer. The optical and photothermal properties of the resulting nanoconstructs (Au-SPNs) are modulated by varying the initial loading input of AuNPs, ranging between 25 and 150 μg_Au. Au-SPNs exhibit a hydrodynamic diameter varying from ~100 to 180 nm, growing with the gold content, and manifest up to 2-fold increase in thermal energy production per unit mass of gold for an initial input of 100 μg_Au. Au-SPNs are stable under physiological conditions up to 7 days and have direct cytotoxic effect on tumor cells. The superior photothermal performance of Au-SPNs is assessed in vitro on monolayers of breast cancer cells (SUM-159) and tumor spheroids of glioblastoma multiforme cells (U87-MG). The encapsulation of small AuNPs into larger spherical nanoconstructs enhances photothermal ablation and could favor tumor accumulation.     

Cellular uptake and radiosensitization of SR-2508 loaded PLGA nanoparticles

SR-2508 (etanidazole), a hypoxic radiosensitizer, has potential applications in radiotherapy. The poly(d,l-lactide-co-glycolide)(PLGA) nanoparticles containing SR-2508 were prepared by w/o/w emulsification-solvent evaporation method. The physicochemical characteristics of the nanoparticles (i.e. encapsulation efficiency, particle size distribution, morphology, in vitro release) were studied. The cellular uptake of the nanoparticles for the two human tumor cell lines: human breast carcinoma cells (MCF-7) and human carcinoma cervices cells (HeLa), was evaluated by fluorescence microscopy and transmission electronic microscopy. Cell viability was measured by the ability of single cell to form colonies in vitro. The prepared nanoparticles were spherical in shape with size between 90 nm and 190 nm. The encapsulation efficiency was 20.06%. The drug release pattern exhibited an initial burst followed by a plateau for over 24 h. The cellular uptake of nanoparticles was observed. Co-culture of MCF-7 and HeLa cells with SR-2508 loaded nanoparticles showed that released SR-2508 retained its bioactivity and effectively sensitized two hypoxic tumor cell lines to radiation. The radiosensitization of SR-2508 loaded nanoparticles was more significant than that of free drug.     

Leveraging re-chargeable nanobubbles on amine-functionalized ZnO nanocrystals for sustained ultrasound cavitation towards echographic imaging

Nanoparticles able to promote inertial cavitation when exposed to focused ultrasound have recently gained much attention due to their vast range of possible applications in the biomedical field, such as enhancing drug penetration in tumor or supporting ultrasound contrast imaging. Due to their nanometric size, these contrast agents could penetrate through the endothelial cells of the vasculature to target tissues, thus enabling higher imaging resolutions than commercial gas-filled microbubbles. Herein, Zinc Oxide NanoCrystals (ZnO NCs), opportunely functionalized with amino-propyl groups, are developed as novel nanoscale contrast agents that are able, for the first time, to induce a repeatedly and over-time sustained inertial cavitation as well as ultrasound contrast imaging. The mechanism behind this phenomenon is investigated, revealing that re-adsorption of air gas nanobubbles on the nanocrystal surface is the key factor for this re-chargeable cavitation. Moreover, inertial cavitation and significant echographic signals are obtained at physiologically relevant ultrasound conditions (MI < 1.9), showing great potential for low side-effects in in-vivo applications of the novel nanoscale agent from diagnostic imaging to gas-generating theranostic nanoplatforms and to drug delivery.     

Carbon Dots‐Cluster‐DOX Nanocomposites Fabricated by a Co‐Self‐Assembly Strategy for Tumor‐Targeted Bioimaging and Therapy

Carbon‐based nanomaterials could afford versatile potential applications in biomedical optical imaging and as nanoparticle drug carriers, owing to their promising optical and biocompatible capabilities. In this paper, it is first found that amphipathic cetylpyridinium chloride (CPC)‐stabilized oil‐soluble carbon dots (CDs) could self‐assemble into hydrophilic CDs clusters with hydrophobic core under ultrasound, in which CPC acts as carbon source, stabilizer, and phase transfer agent. Next, the size‐control (for size‐dependent passive tumor targeting) and doxorubicin (DOX) uploading of aqueous CDs clusters, and subsequent surface charge modification via overcoating with cRGD‐ and octylamine‐modified polyacrylic acid (cRGD‐PAA‐OA) (reversing their surface charges into negative and introducing active tumor‐targeting ability) are explored systematically. Based on this sequential administration mode, CDs‐cluster‐DOX/cRGD‐PAA‐OA nanocomposites exhibit selective human malignant glioma cell line (U87MG) tumor targeting. In in vitro drug release experiments, the nanocomposites could release DOX timely. Owning to the dual tumor targeting effects and seasonable drug release, CDs‐cluster‐DOX/cRGD‐PAA‐OA show remarkably tumor targetability and enhanced antitumor efficacy (and reduced adverse reaction), comparing to free DOX in animal models. These results indicate that fabricating nanocomposite via co‐self‐assembly strategy is efficient toward drug delivery system for tumor‐targeting theranostic.     

Bioapplications of poly(amidoamine) (PAMAM) dendrimers in nanomedicine

Poly(amidoamine) (PAMAM) dendrimers are a novel class of spherical, well-designed branching polymers with interior cavities and abundant terminal groups on the surface which can form stable complexes with drugs, plasmid DNA, oligonucleotides, and antibodies. Amine‐terminated PAMAM dendrimers are able to solubilize different families of hydrophobic drugs, but the cationic charges on dendrimer surface may disturb the cell membrane. Therefore, surface modification by PEGylation, acetylation, glycosylation, and amino acid functionalization is a convenient strategy to neutralize the peripheral amine groups and improve dendrimer biocompatibility. Anticancer agents can be either encapsulated in or conjugated to dendrimer and be delivered to the tumor via enhanced permeability and retention (EPR) effect of the nanoparticle and/or with the help of a targeting moiety such as antibody, peptides, vitamins, and hormones. Biodegradability, non-toxicity, non-immunogenicity, and multifunctionality of PAMAM dendrimer are the key factors which facilitate steady increase of its application in drug delivery, gene transfection, tumor therapy, and diagnostics applications with precision and selectivity. This review deals with the major topics of PAMAM dendrimers including structure, synthesis, toxicity, surface modification, and also possible new applications of these spherical polymers in biomedical fields as dendrimer-based nanomedicine.     

Role of spin-orbit splitting and dynamical fluctuations in the Si(557)-Au surface

Our ab initio calculations show that spin-orbit coupling is crucial to understand the electronic structure of the Si(557)-Au surface. The spin-orbit splitting produces the two one-dimensional bands observed in photoemission, which were previously attributed to spin-charge separation in a Luttinger liquid. This spin splitting might have relevance for future device applications. We also show that the apparent Peierls-like transition observed in this surface by scanning tunneling microscopy is a result of the dynamical fluctuations of the step-edge structure which are quenched as the temperature is decreased.     

A Functionalized Hollow Mesoporous Silica Nanoparticles‐Based Controlled Dual‐Drug Delivery System for Improved Tumor Cell Cytotoxicity

Multifunctional nanoparticles for selectively targeting tumor cells and effectively delivering multiple drugs are urgently needed in cancer therapy. Here, a dual‐drug delivery system is prepared, based on functionalized hollow mesoporous silica nanoparticles (HMSNs). Doxorubicin (DOX) hydrochloride is loaded into the hollow core, and dichloro(1,2‐diaminocyclohexane)platinum (II) (DACHPt) is stored in the pores of the shell by the coordination interaction with the carboxyl groups modified on the pore walls, which also serves as barriers to control the DOX release. Detailed studies in vitro indicate that the DACHPt release is triggered by Cl^? through the cleavage of the coordination interaction, and the DOX release depends on the release rate of DACHPt and the environmental pH value. The surface of the mechanized nanoparticles is also modified by transferrin (Tf) to achieve the tumor specificity. Compared with individual drug delivery systems, the dual‐drug delivery system shows synergistic efficacy on the cell cytotoxicity (combination index = 0.30), resulting in improved tumor cell killing. The present dual‐drug delivery system provides a promising strategy to develop controlled and targeted combination therapies for efficient cancer treatment.     

Functionalized magnetic nanoparticles for drug delivery in tumor therapy

The side effects of chemotherapy are mainly the poor control of drug release. Magnetic nanoparticles(MNPs) have super-paramagnetic behaviors which are preferred for biomedical applications such as in targeted drug delivery, besides, in magnetic recording, catalysis, and others. MNPs, due to high magnetization response, can be manipulated by the external magnetic fields to penetrate directly into the tumor, thus they can act as ideal drug carriers. MNPs also play a crucial role in drug delivery system because of their high surface-to-volume ratio and porosity. The drug delivery in tumor therapy is related to the sizes, shapes, and surface coatings of MNPs as carriers. Therefore, in this review, we first summarize the effects of the sizes, shapes, and surface coatings of MNPs on drug delivery, then discuss three types of drug release systems, i.e., p H-controlled, temperature-controlled, and magnetic-controlled drug release systems, and finally compare the principle of passive drug release with that of active drug release in tumor therapy.     

Identification of novel antitumor agents from mixture-based synthetic combinatorial libraries using cell-based assays

A new strategy is presented here which integrates combinatorial library technology with the antitumor in vitro screening system at the National Cancer Institute in the search for novel antitumor agents. Mixture-based synthetic combinatorial libraries (SCLs) representing hundreds of thousands to millions of individual compounds were screened against the cell-based assay, which evaluates compounds for their ability to inhibit the growth of 60 different human tumor cell lines. Five different SCLs, composed of peptides, peptidomimetics, polyamines or small molecules were first tested against three cell lines to identify the most active SCLs. Two SCLs, namely the N-perbenzylated pentamine and the N-acylated permethylated triamine, were deconvoluted to yield individual compounds having significant activities against the 60 tumor cell lines. Active compounds were tested in mice to determine the maximum tolerated dose, followed by in vivo testing in a hollow fiber assay. Using this strategy, three different compounds identified directly from SCLs are currently being evaluated in human tumor xenografts. This study demonstrates for the first time the use of in vitro cell-based assays to identify antitumor lead compounds from mixture-based combinatorial libraries.     

Ultrasound-driven titanium modification with formation of titania based nanofoam surfaces

Titanium has been widely used as biomaterial for various medical applications because of its mechanical strength and inertness. This on the other hand makes it difficult to structure it. Nanostructuring can improve its performance for advanced applications such as implantation and lab-on-chip systems. In this study we show that a titania nanofoam on titanium can be formed under high intensity ultrasound (HIUS) treatment in alkaline solution. The physicochemical properties and morphology of the titania nanofoam are investigated in order to find optimal preparation conditions for producing surfaces with high wettability for cell culture studies and drug delivery applications. AFM and contact angle measurements reveal, that surface roughness and wettability of the surfaces depend nonmonotonously on ultrasound intensity and duration of treatment, indicating a competition between HIUS induced roughening and smoothening mechanisms. We finally demonstrate that superhydrophilic bio-and cytocompatible surfaces can be fabricated with short time ultrasonic treatment.     

Influence of size and surface capping on photoluminescence and cytotoxicity of gold nanoparticles

Hydrophilic and homogeneous sub-10 nm blue light-emitting gold nanoparticles (NPs) functionalized with different capping agents have been prepared by simple chemical routes. Structure, average, size, and surface characteristics of these NPs have been widely studied, and the stability of colloidal NP solutions at different pH values has been evaluated. Au NPs show blue PL emission, particularly in the GSH capped NPs, in which the thiol-metal core transference transitions considerably enhance the fluorescent emission. The influence of capping agent and NP size on cytotoxicity and on the fluorescent emission are analyzed and discussed in order to obtain Au NPs with suitable features for biomedical applications. Cytotoxicity of different types of gold NPs has been determined using NPs at high concentrations in both tumor cell lines and primary cells. All NPs used show high biocompatibility with low cytotoxicity even at high concentration, while Au-GSH NPs decrease viability and proliferation of both a tumor cell line and primary lymphocytes.     

The effect of dexamethasone on tissue water distribution and proton relaxation in Panc02 tumors

The present experiments were conducted to determine the effects of dexamethasone mediated changes in tumor water distribution on proton relaxation times (T1, T2) in a murine pancreatic adenocarcinoma (Panc02). Spin lattice (T1) and spin-spin (T2) relaxation times were determined by ex vivo methods (10MHz) and by in vivo imaging techniques (6.25 MHz) at various intervals after single or multiple dexamethasone treatments. In complementary studies, dexamethasone mediated changes in tumor capillary permeability, tumor water distribution, relative tumor blood flow and tumor cell proliferation were also determined.

Proton spin lattice (T1) and spin-spin (T2 relaxation times for Panc02 tumors shortened within two hours of a single dexamethasone treatment. The time course and magnitude of this response was dexamethasone dose dependent. The time dependent changes in T1 and T2 after dexamethasone were similar at 10 MHz (ex vivo) and 6.25 MHz (in vivo imaging). Although dexamethasone produced little or no change in total tumor water content and tumor cell proliferation, transient changes in the physiologic distribution of tumor water were clearly demonstrated.

The data supports the idea that dexamethasone induced changes in the distribution of tumor water were mediated by changes in capillary permeability and tumor blood flow. These physiologic responses produced serial changes in tumor extracellular extravascular water content that were consistent with the observed changes in tumor T1 and T2. The results from these experiments might imply that therapy associated changes in tumor proton relaxation times may not only reflect changes in tissue water content, but may also reflect physiologic responses which alter the distribution of tissue water and solute.     

基于金纳米粒子的光谱分析法进行单细胞探测

在过去的几十年中,等离子纳米粒子,尤其是金纳米粒子(AuNPs),由于其独特的局部表面等离子体共振(LSPR)特性,金纳米颗粒非常适合高度传导定域在表面的化学或物理刺激产生的光信号,已被广泛应用于生物检测与成像。包括单细胞光谱分析与成像。基于光吸收和弹性光学方法散射,阐述了利用光谱方法进行的单细胞光学探测的进展应用和纳米系统表现出新的特性。论述了基于AuNPs的细胞内环境光谱分析与探测,对在单细胞水平上进行的细胞动态实时测量的基本原则和与光互动独特的相关方法进行了描述。重点放在单细胞光谱检测的原则、方法及这些方法的优点和挑战,并阐述了最近在这一领域的研究进展,内容包括细胞和亚细胞环境的探测、细胞应答诱导细胞凋亡过程探测、生物分子识别和量化、药物传递及释放、癌症诊断及治疗等。给出了未来的挑战和努力方向。     

Modeling of various contact theories for the manipulation of different biological micro/nanoparticles based on AFM

Single-walled carbon nanohorns (SWNHs) have great potential to enhance thermal and chemotherapeutic drug efficiencies for cancer therapies. Despite their diverse capabilities, minimal research has been conducted so far to study nanoparticle intracellular transport, which is an important step in designing efficient therapies. SWNHs, like many other carbon nanomaterials, do not have inherent fluorescence properties making intracellular transport information difficult to obtain. The goals of this project were to (1) develop a simple reaction scheme to decorate the exohedral surface of SWNHs with fluorescent quantum dots (QDs) and improve conjugate stability, and (2) evaluate SWNH–QD conjugate cellular uptake kinetics and localization in various cancer cell lines of differing origins and morphologies. In this study, SWNHs were conjugated to CdSe/ZnS core/shell QDs using a unique approach to carbodiimide chemistry. Transmission electron microscopy and electron dispersive spectroscopy verified the conjugation of SWNHs and QDs. Cellular uptake kinetics and efficiency were characterized in three malignant cell lines: U-87 MG (glioblastoma), MDA-MB-231 (breast cancer), and AY-27 (bladder transitional cell carcinoma) using flow cytometry. Cellular distribution was verified by confocal microscopy, and cytotoxicity was also evaluated using an alamarBlue assay. Results indicate that cellular uptake kinetics and efficiency are highly dependent on cell type, highlighting the significance of studying nanoparticle transport at the cellular level. Nanoparticle intracellular transport investigations may provide information to optimize treatment parameters (e.g., SWNH concentration, treatment time, etc.) depending on tumor etiology.     

Effect of geometry on impedance of cell suspended media in electrically mediated molecule uptake using a microstructure

A systematic design and miniaturization of conventional clinical and diagnostic tools are emerging in many nanobiotechnology and nanobioscience fields. Recently, the pharmaceutical and biotechnological industries have continuously accelerated novel drug discovery for cellular therapeutics of hard-to-cure diseases. Electroporation is a promising candidate in gene transfection methods for future cellular therapeutics. Here, we report our initial study on the effect of geometry on impedance of cell suspended media in electroporation using a microchannel. A full understanding of the electroporation environment and conditions enabled us to optimize protocols of electroporation for various cell lines efficiently. Furthermore, this study will open up a new vista of future electroporation devices using microstructures.     

基于NMR的绒癌化疗耐药机制的代谢组学研究

绒癌是一种发生于育龄妇女的恶性滋养细胞肿瘤.经甲氨蝶呤、5-氟尿嘧啶等单药或多药联合化疗后,其治愈率通常高达90%.然而耐药仍是绒癌治疗失败的主要原因.目前绒癌耐药的机制尚不明确,前人多从蛋白和转录水平阐述,鲜有涉及代谢物层次的研究.本文采用基于核磁共振（NMR）的代谢组学方法,研究了甲氨蝶呤耐药株JEG3R与敏感株JEG3绒癌细胞及其培养基在代谢物水平上的差异.结果发现JEG3R耐药极可能与糖代谢异常相关,主要表现为耐药株细胞中山梨糖醇的累积,提示醛糖还原酶可能在耐药过程中起重要作用;其次,耐药株细胞中乳酸积累、丙氨酸流入三羧酸循环途径增强;此外,耐药株细胞相比敏感株细胞中氨基酸、核苷酸及磷酸胆碱等必需营养物质的显著性改变,且其变化与绒癌增殖速率正相关.本研究为绒癌的耐药机制研究补充了代谢层次的数据,为后续从分子生物学水平上揭示耐药机制提供了新思路.     

Improving cancer treatments via dynamical biophysical models

Despite significant advances in oncological research, cancer nowadays remains one of the main causes of mortality and morbidity worldwide. New treatment techniques, as a rule, have limited efficacy, target only a narrow range of oncological diseases, and have limited availability to the general public due their high cost. An important goal in oncology is thus the modification of the types of antitumor therapy and their combinations, that are already introduced into clinical practice, with the goal of increasing the overall treatment efficacy. One option to achieve this goal is optimization of the schedules of drugs administration or performing other medical actions. Several factors complicate such tasks: the adverse effects of treatments on healthy cell populations, which must be kept tolerable; the emergence of drug resistance due to the intrinsic plasticity of heterogeneous cancer cell populations; the interplay between different types of therapies administered simultaneously. Mathematical modeling, in which a tumor and its microenvironment are considered as a single complex system, can address this complexity and can indicate potentially effective protocols, that would require experimental verification. In this review, we consider classical methods, current trends and future prospects in the field of mathematical modeling of tumor growth and treatment. In particular, methods of treatment optimization are discussed with several examples of specific problems related to different types of treatment.     

Overcoming the rayleigh criterion limit with optical vortices

We experimentally and numerically tested the separability of two independent equally luminous monochromatic and white light sources at the diffraction limit, using optical vortices (OV). The diffraction pattern of one of the two sources crosses a fork hologram on its center generating the Laguerre-Gaussian (LG) transform of an Airy disk. The second source, crossing the fork hologram in positions different from the optical center, generates nonsymmetric LG patterns. We formulated a criterion, based on the asymmetric intensity distribution of the superposed LG patterns so created, to resolve the two sources at angular distances much below the Rayleigh criterion. Analogous experiments in white light allow angular resolutions which are still one order of magnitude below the Rayleigh criterion. The use of OVs might offer new applications for stellar separation in future space experiments.     

The engineering of doxorubicin-loaded liposome-quantum dot hybrids for cancer theranostics

In the fabrication of phase change random access memory(PRAM) devices, high temperature thermal processes are inevitable. We investigate the thermal stability of Ge2Sb2Te5(GST) which is a prototypical phase change material. After high temperature process, voids of phase change material exist at the interface between Ge2Sb2Te5 and substrate in the initial open memory cell. This lower region of Ge2Sb2Te5 is found to be a Te-rich phase change layer. Phase change memory devices are fabricated in different process conditions and examined by scanning electron microscopy and energy dispersive X-ray. It is found that hot-chuck process, nitrogen-doping process, and lower temperature inter-metal dielectric(IMD) deposition process can ease the thermal impact of line-GST PRAM cell.