Zheyu Shen

Preparation of Hierarchical Hollow CaCO3 Particles and the Application as Anticancer Drug Carrier

One-pot approach to couple the crystallization of CaCO(3) nanoparticles and the in situ symmetry-breaking assembly of these crystallites into hollow spherical shells was developed under the templating effect of a soluble starch. Further functional study using HP-a as an anticancer drug carrier (DOX) demonstrated its advantages for localizing drug release by the pH value-sensitive structure and enhancing cytotoxicity by increasing cellular uptake, perinuclear accumulation, and nuclear entry.

Iron Oxide Nanoparticle Based Contrast Agents for Magnetic Resonance Imaging

Magnetic iron oxide nanoparticles (MIONs) have attracted enormous attention due to their wide applications, including for magnetic separation, for magnetic hyperthermia, and as contrast agents for magnetic resonance imaging (MRI). This review article introduces the methods of synthesizing MIONs, and their application as MRI contrast agents. Currently, many methods have been reported for the synthesis of MIONs. Herein, we only focus on the liquid-based synthesis methods including aqueous phase methods and organic phase methods. In addition, the MIONs larger than 10 nm can be used as negative contrast agents and the recently emerged extremely small MIONs (ES-MIONs) smaller than 5 nm are potential positive contrast agents. In this review, we focus on the ES-MIONs because ES-MIONs avoid the disadvantages of MION-based T2- and gadolinium chelate-based T1-weighted contrast agents.

Improved drug targeting of cancer cells by utilizing actively targetable folic acid-conjugated albumin nanospheres

Folic acid-conjugated albumin nanospheres (FA-AN) have been developed to provide an actively targetable drug delivery system for improved drug targeting of cancer cells with reduced side effects. The nanospheres were prepared by conjugating folic acid onto the surface of albumin nanospheres using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC) as a catalyst. To test the efficacy of these nanospheres as a potential delivery platform, doxorubicin-loaded albumin nanospheres (DOX-AN) and doxorubicin-loaded FA-AN (FA-DOX-AN) were prepared by entrapping DOX (an anthracycline, antibiotic drug widely used in cancer chemotherapy that works by intercalating DNA) into AN and FA-AN nanoparticles. Cell uptake of the DOX was then measured. The results show that FA-AN was incorporated into HeLa cells (tumor cells) only after 2.0h incubation, whereas HeLa cells failed to incorporate albumin nanospheres without conjugated folic acid after 4.0h incubation. When HeLa cells were treated with the DOX-AN, FA-DOX-AN nanoparticles or free DOX, cell viability decreased with increasing culture time (i.e. cell death increases with time) over a 70h period. Cell viability was always the lowest for free DOX followed by FA-DOX-AN4 and then DOX-AN. In a second set of experiments, HeLa cells washed to remove excess DOX after an initial incubation for 2h were incubated for 70h. The corresponding cell viability was slightly higher when the cells were treated with FA-DOX-AN or free DOX whilst cells treated with DOX-AN nanoparticles remained viable. The above experiments were repeated for non-cancerous, aortic smooth muscle cells (AoSMC). As expected, cell viability of the HeLa cells (with FA receptor alpha, FRα) and AoSMC cells (without FRα) decreased rapidly with time in the presence of free DOX, but treatment with FA-DOX-AN resulted in selective killing of the tumor cells. These results indicated that FA-AN may be used as a promising actively targetable drug delivery system to improve drug targeting to cancer cells.

Thermosensitive polymer-conjugated albumin nanospheres as thermal targeting anti-cancer drug carrier

Thermosensitive Poly(N-isopropylacrylamide-co-acrylamide-co-allylamine) (PNIPAM-AAm-AA)-conjugated albumin nanospheres (PAN) was developed as a new thermal targeting anti-cancer drug carrier by conjugating PNIPAM-AAm-AA on the surface of albumin nanospheres (AN). AN with diameter below 200nm and narrow size distribution was successfully prepared in the first step with desolvation technique. PNIPAM-AAm-AA with different molecular weight (M(w)) was synthesized in the second step by radical polymerization and conjugated onto the surface of AN. Anti-cancer drug adriamycin (ADR) was then entrapped into the AN and PAN during the particle preparation. Compared with AN, the release rate of ADR from PAN in trypsin solution was slower, and decreased with increasing the conjugation amounts (hairy density) or M(w) of PNIPAM-AAm-AA (hairy length). Moreover, the release of ADR from PAN above the cloud-point temperature (T(cp)) of PNIPAM-AAm-AA became faster due to shrinkage of hairy thermosensitive polymer. To testify the thermal targetability in vivo, PAN was incubated with HepG2 cells. As expected, PAN can target cancer cells above the T(cp) of PNIPAM-AAm-AA, whereas it cannot below the T(cp). These results might reflect that PAN may selectively accumulate onto solid tumors that are maintained above physiological temperature due to local hyperthermia.

Exploring N-Imidazolyl-O-Carboxymethyl Chitosan for High Performance Gene Delivery

Chitosan shows good biocompatibility and biodegradability, but the poor water solubility and low transfection efficiency hinder its applications as a gene delivery vector. We here report the detailed synthesis and characterization of a novel ampholytical chitosan derivative, N-imidazolyl-O-carboxymethyl chitosan (IOCMCS), used for high performance gene delivery. After chemical modification, the solubility of the resulting polymer is enhanced, and the polymer is soluble in a wide pH range (4-10). Gel electrophoresis study reveals the strong binding ability between plasmid DNA and the IOCMCS. Moreover, the IOCMCS does not induce remarkable cytotoxicity against human embryonic kidney (HEK293T) cells. The cell transfection results with HEK293T cells using the IOCMCS as gene delivery vector demonstrate the high transfection efficiency, which is dependent on the degree of imidazolyl substitution. Therefore, the IOCMCS is a promising candidate as the DNA delivery vector in gene therapy due to its high solubility, high gene binding capability, low cytotoxicity, and high gene transfection efficiency.

A galactosamine-mediated drug delivery carrier for targeted liver cancer therapy

In order to minimize the side effect of cancer chemotherapy, a novel galactosamine-mediated drug delivery carrier, galactosamine-conjugated albumin nanoparticles (GAL-AN), was developed for targeted liver cancer therapy. The albumin nanoparticles (AN) and doxorubicin-loaded AN (DOX-AN) were prepared by the desolvation of albumin in the presence of glutaraldehyde crosslinker. Morphological study indicated the spherical structure of these synthesized particles with an average diameter of around 200 nm. The functional ligand of galactosamine (GAL) was introduced onto the surfaces of AN and DOX-AN via carbodiimide chemistry to obtain GAL-AN and GAL-DOX-AN. Cellular uptake and kinetic studies showed that GAL-AN is able to be selectively incorporated into the HepG2 cells rather than AoSMC cells due to the existence of asialoglycoprotein receptors on HepG2 cell surface. The cytotoxicity, measured by MTT test, indicated that AN and GAL-AN are non-toxic and GAL-DOX-AN is more effective in HepG2 cell killing than that of DOX-AN. As such, our results implied that GAL-AN and GAL-DOX-AN have specific interaction with HepG2 cells via the recognition of GAL and asialoglycoprotein receptor, which renders GAL-AN a promising anticancer drug delivery carrier for liver cancer therapy.

Colorimetric response of dithizone product and hexadecyl trimethyl ammonium bromide modified gold nanoparticle dispersion to 10 types of heavy metal ions: understanding the involved molecules from experiment to simulation.

A new kind of analytical reagent, hexadecyl trimethyl ammonium bromide (CTAB), and dithizone product-modified gold nanoparticle dispersion, is developed for colorimetric response to 10 types of heavy metal ions (M(n+)), including Cr(VI), Cr(3+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+), and Pb(2+). The color change of the modified gold nanoparticle dispersion is instantaneous and distinct for Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+), and Pb(2+). The color change results from the multiple reasons, such as electronic transitions, cation-π interactions, formation of coordination bonds, and M(n+)-induced aggregation of gold nanoparticles (AuNPs). The different combining capacity of heavy metal ions to modifiers results in the different broadening and red-shifting of the plasmon peak of modified AuNPs. In addition, Cr(VI), Cu(2+), Co(2+), Ni(2+), and Mn(2+) cause the new UV-vis absorption peaks in the region of 360-460 nm. The interactions between the modifiers and AuNPs, and between the modifiers and M(n+), are investigated by using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The results confirm that AuNPs are modified by CTAB and dithizone products through electrostatic interactions and Au-S bonds, respectively, and the M(n+)-N bonds form between M(n+) and dithizone products. Furthermore, the experimental and density functional theory calculated IR spectra prove that dithizone reacts with NaOH to produce C6H5O(-) and [SCH2N4](2-). The validation of this method is carried out by analysis of heavy metal ions in tap water.

Improved SERS Nanoparticles for Direct Detection of Circulating Tumor Cells in the Blood

The detection of circulating tumor cells (CTCs) in the blood of cancer patients is crucial for early cancer diagnosis, cancer prognosis, evaluation of the treatment effect of chemotherapy drugs, and choice of cancer treatment options. In this study, we propose new surface-enhanced Raman scattering (SERS) nanoparticles for the direct detection of CTCs in the blood. Under the optimized experimental conditions, our SERS nanoparticles exhibit satisfying performances for the direct detection of cancer cells in the rabbit blood. A good linear relationship is obtained between the SERS intensity and the concentration of cancer cells in the range of 5-500 cells/mL (R(2) = 0.9935), which demonstrates that the SERS nanoparticles can be used for the quantitative analysis of cancer cells in the blood and the limit of detection is 5 cells/mL, which is lowest compared with the reported values. The SERS nanoparticles also have an excellent specificity for the detection of cancer cells in the rabbit blood. The above results reinforce that our SERS nanoparticles can be used for the direct detection of CTCs in the blood with excellent specificity and high sensitivity.

Exploring a new SPION-based MRI contrast agent with excellent water-dispersibility, high specificity to cancer cells and strong MR imaging efficacy

Advances in contrast agents have greatly enhanced the sensitivity of magnetic resonance imaging (MRI) technique for early diagnosis of cancer. However, the commercial superparamagnetic iron oxide nanoparticles (SPION)-based contrast agents synthesized by co-precipitation method are not monodisperse with irregular morphologies and ununiform sizes. Other reported SPION-based contrast agents synthesized by solvothermal method or thermal decomposition method are limited by the bad water-dispersibility and low specificity to cancer cells. Herein, we propose a new strategy for exploring SPION-based MRI contrast agents with excellent water-dispersibility and high specificity to cancer cells. The SPION was synthesized by a polyol method and then entrapped into albumin nanospheres (AN). After that, a ligand folic acid (FA) was conjugated onto the surface of the AN to construct a SPION-AN-FA composite. The transmission electron microscope (TEM) and dynamic light scattering (DLS) results indicate that the SPION-AN-FA has a spherical shape, a uniform size and an excellent water-dispersibility (polydispersity index (PDI) <0.05). The results of laser scanning confocal microscope (LSCM) and flow cytometry demonstrate that the SPION-AN-FA nanoparticles are highly specific to MCF-7 and SPC-A-1 cells due to the recognition of ligand FA and folate receptor α (FRα). The r2/r1 value of SPION-AN-FA is around 40, which is much higher than that of Resovist(®) indicating that our SPION-AN-FA has a stronger T2 shortening effect. The T2-weighted images of MCF-7 cells incubated with SPION-AN-FA are significantly darker than those of MCF-7 cells incubated with AN, indicating that our SPION-AN-FA has a strong MR imaging efficacy. In view of the excellent water-dispersibility, the high specificity to cancer cells and the strong MR imaging efficacy, our SPION-AN-FA can be used as a negative MR contrast agent.

Developing a chitosan supported imidazole Schiff-base for high-efficiency gene delivery

A chitosan supported imidazole Schiff-base (CISB) has been developed to be used as the vector for high performance gene delivery. Introducing the imidazole Schiff-base to the branch of chitosan could improve its water solubility and gene binding ability under physiological conditions, and thus significantly enhance gene delivery capability due to the formation of Schiff-bases (azomethines) and the substitution of imidazole functional groups along chitosan backbones. Gel electrophoresis and light scattering results show that the CISB could effectively bind plasmid DNA (pDNA) and protect pDNA from DNase I digestion in solution. The CISB does not induce remarkable cytotoxicity against HEK 293 cells and can enhance delivery of pDNA into cytoplasm and nucleus efficiently. A transfection efficiency of 70% can be reached after systematically optimizing cell transfection conditions. Therefore, the CISB is a promising gene delivery vector due to its high solubility in physiological pH, strong gene binding and protection capability, low cytotoxicity, good biodegradability, and high efficiency in gene delivery and cell transfection.