ARGININE- AND ACRYLONITRILE-MODIFIED CHITOSAN NANOPARTICLES FOR ANTICANCER DRUG DELIVERY

LIANCHENG YIN,Zhongwei Gu,TING SU,JING CHANG,Bin He,RONG LIU 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2014 NANO Vol.9 No.7

Chitosan (CS) is an excellent natural biodegradable and biocompatible polymer for biomedicalapplications, however, its poor solubility in water or organic solvents limits its applications indrug delivery. In order to resolve this problem, chitosan was modi¯ed with acrylonitrile (AN)and arginine (Arg), the modi¯ed chitosan (AN – CS – Arg) was characterized by1H NMR andFourier transform infrared (FTIR). The AN – CS – Arg was self-assembled into nanoparticles toencapsulate anticancer drug doxorubicin (DOX). The size and morphology of the blank anddrug-loaded AN – CS – Arg (AN – CS – Arg/DOX) nanoparticles were measured by dynamic lightscattering (DLS), scanning electron microscopy (SEM) and atomic force microscopy (AFM). Themean size of both blank and AN – CS – Arg/DOX nanoparticles were around 50 nm and 170 nm,respectively. The drug-loading content was about 12%. The release pro¯le of AN – CS – Arg/DOXnanoparticles was investigated in vitro, 80% encapsulated DOX could be released within 80 h. The AN – CS – Arg nanoparticles were nontoxic to both NIH 3T3 ¯broblasts and HepG2 cancercells. The cellular uptake of the AN – CS – Arg nanoparticles was tra±cked via Confocal LaserScanning Microscopy and Flow Cytometry, both results showed that the AN – CS – Arg nano-particles could be internalized in HepG2 cells e±ciently. The IC 50 of AN – CS – Arg/DOX nano-particles to HepG2 cancer cells was 10.0 ? g/mL. The AN – CS – Arg nanoparticles are potentialcarriers for anticancer drug delivery.

Synthesis, Characterization, and Drug Delivery of Amphiphilic Poly{(lactic acid)-co-[(glycolic acid)-alt-(L-glutamic acid)]}-g-Poly(ethylene glycol)

Zuxiao Yu,Zhongwei Gu,Bin He,Chunyan Long,Rong Liu,Mingming Sheng,Gang Wang,James. Z. Tang 한국고분자학회 2012 Macromolecular Research Vol.20 No.3

This paper discusses the use of a novel amphiphilic graft polymer poly{(lactic acid)-co-[(glycolic acid)-alt-(Lglutamic acid)]}-g-monomethyl poly(ethylene glycol) (PLGG-g-mPEG) as a drug carrier. PLGG was synthesized through the ring-opening polymerization of L-lactide (LLA) and (3s)-benzoxylcarbonylethyl-morpholine-2,5-dione (BEMD) using Sn(Oct)2 as a catalyst and it was subsequently deprotected via hydrogenolysis in the presence of Pd/C. A series of monomethyl poly(ethylene glycol) (PEG) with the molecular weights of 2,000, 1,100, and 500 were immobilized on the carboxyl groups of PLGG. These PEGylated graft derivatives were characterized using proton nuclear magnetic resonance spectra (1H NMR), Fourier transform infrared spectroscopy (FTIR), and gel permeation chromatography (GPC). The critical micelle concentrations (CMCs) of the amphiphilic copolymers were tested by the fluorescence probe technique and the CMCs were 2.3, 1.0, and 0.32 μg/mL, respectively. Transmission electronic microscopy (TEM) and dynamic light scattering (DLS) images revealed that the micelles were homogeneous spherical nanoparticles and the sizes of the micelles were distributed across a range of 80 to 22 nm. Anticancer drug doxorubicin (DOX) was loaded into the micelles. The in vitro release profiles showed that the sustaining release of the drugloaded micelles could last over 7 days. The in vitro cytotoxicity assay of the DOX-loaded micelles against HepG2cells was assessed by methyl thiazolyl tetrazolium (MTT) assays. The results demonstrated that the drug-loaded micelles exhibited a high level of inhibition activity on cancer cells. The confocal microscopy images of HepG2 cells showed that DOX released from the micelles could be delivered into cell nuclei. PLGG-g-mPEG micelles are promising potential carriers for delivering anticancer drugs.

Protein corona on magnetite nanoparticles and internalization of nanoparticle-protein complexes into healthy and cancer cells

Jiang, Wen,Lai, Kuilin,Wu, Yao,Gu, Zhongwei 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.1

Superparamagnetic magnetite nanoparticles (MNPs) of different surface properties are incubated in complicated living fluid, including fetal bovine serum solution, cell complete culture medium and cell culture system with/without serum, to investigate the alteration of protein corona and its impact on cell internalization. The MNPs prepared by co-precipitation method are functionalized with L-Lysine (Lys), Glucosamic acid (GA) to obtain amine, carboxyl and hydroxyl groups, separately. All the particles adsorb serum proteins to form MNPs-protein complexes with the surface charge changing into negative. 1D SDS/PAGE gel images analysis indicates that the composition and content of hard protein corona on the surface of NPs are related to their functional groups and agglomeration, and the total amount of protein in the medium. In cell culture system, particles not only adsorb serum proteins, but also associate with cytosolic proteins arising from HepG2 and L02 cells. GA modified MNPs (MNPs-GA) exhibit bovine serum albumin anti-adsorption capability because of the terminal hydroxyl and carboxyl groups. MNPs-GA also shows the highest cellular uptake and label efficiency compared with uncoated MNPs and Lys modified MNPs, due to larger aggregates formation and specific protein corona composition, rather than commonly approved electrostatic interaction between particles and cells. For the first time, our results provide visualized reports on previously neglected, but indispensable protein corona of the MNPs after interaction with both healthy and cancer cells, suggesting that cytosolic protein corona from cells and aggregation of particles are important factors needed to be account for on studying the nano-bio interface.

Protein corona on magnetite nanoparticles and internalization of nanoparticle–protein complexes into healthy and cancer cells

Wen Jiang,Kuilin Lai,Yao Wu,Zhongwei Gu 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.1

Superparamagnetic magnetite nanoparticles(MNPs) of different surface properties are incubated incomplicated living fluid, including fetal bovine serumsolution, cell complete culture medium and cell culturesystem with/without serum, to investigate the alteration ofprotein corona and its impact on cell internalization. TheMNPs prepared by co-precipitation method are functionalizedwith L-Lysine (Lys), Glucosamic acid (GA) to obtainamine, carboxyl and hydroxyl groups, separately. All theparticles adsorb serum proteins to form MNPs–proteincomplexes with the surface charge changing into negative. 1D SDS/PAGE gel images analysis indicates that thecomposition and content of hard protein corona on thesurface of NPs are related to their functional groups andagglomeration, and the total amount of protein in themedium. In cell culture system, particles not only adsorbserum proteins, but also associate with cytosolic proteinsarising from HepG2 and L02 cells. GA modified MNPs(MNPs-GA) exhibit bovine serum albumin anti-adsorptioncapability because of the terminal hydroxyl and carboxylgroups. MNPs-GA also shows the highest cellular uptakeand label efficiency compared with uncoated MNPs andLys modified MNPs, due to larger aggregates formationand specific protein corona composition, rather than commonlyapproved electrostatic interaction between particles and cells. For the first time, our results provide visualizedreports on previously neglected, but indispensable proteincorona of the MNPs after interaction with both healthy andcancer cells, suggesting that cytosolic protein corona fromcells and aggregation of particles are important factorsneeded to be account for on studying the nano–biointerface.

Preparation of monodisperse magnetite nanoparticles under mild conditions

Xiantao Wen,Junxiao Yang,Bin He,Zhongwei Gu 한국물리학회 2008 Current Applied Physics Vol.8 No.5

In this paper, we reported a method to prepare monodisperse magnetite nanoparticles at mild temperature using cheap and non-toxic precursors. It overcomes the shortages of chemical co-precipitation method and thermal decomposition method and combines the advantages of facile, cheap, large-scale, monodisperse, nanosize, and low synthesis temperature and low toxic. In this method, FeCl₃ㆍ6H₂O, FeCl₂ㆍ4H₂O and sodium oleate were mixed in toluene/ethanol/water mixture solvent and refluxed at 74℃ to prepare magnetite nanoparticles directly. The nanoparticles were characterized by transmission electron microscopy, dynamic light scattering, X-ray diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectrometer and thermogravimetric analysis. The magnetic properties of nanoparticles were measured by superconducting quantum interference device. The results showed that the nanoparticles are well-monodisperse with about 4–5 nm of average diameter. The nanoparticles were proved to be superparamagnetic with saturated magnetization 23.6 emu/g and blocking temperature 24.4 K. A possible formation mechanism of monodisperse magnetite nanoparticles was presented at the same time. In this paper, we reported a method to prepare monodisperse magnetite nanoparticles at mild temperature using cheap and non-toxic precursors. It overcomes the shortages of chemical co-precipitation method and thermal decomposition method and combines the advantages of facile, cheap, large-scale, monodisperse, nanosize, and low synthesis temperature and low toxic. In this method, FeCl₃ㆍ6H₂O, FeCl₂ㆍ4H₂O and sodium oleate were mixed in toluene/ethanol/water mixture solvent and refluxed at 74℃ to prepare magnetite nanoparticles directly. The nanoparticles were characterized by transmission electron microscopy, dynamic light scattering, X-ray diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectrometer and thermogravimetric analysis. The magnetic properties of nanoparticles were measured by superconducting quantum interference device. The results showed that the nanoparticles are well-monodisperse with about 4–5 nm of average diameter. The nanoparticles were proved to be superparamagnetic with saturated magnetization 23.6 emu/g and blocking temperature 24.4 K. A possible formation mechanism of monodisperse magnetite nanoparticles was presented at the same time.

Preparation and Gene Delivery of Alkaline Amino Acids-Based Cationic Liposomes

Li, Li,Nie, Yu,Zhu, Rong,Shi, Sanjun,Luo, Kui,He, Bin,Yang, Yang,Yang, Junxiao,Gu, Zhongwei 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.7

Cationic lipids 1, 2, and 3, based on hydrophobic cholesterol linked to L-lysine, L-histidine or Larginine, respectively, were designed and tested as gene delivery vectors. Physicochemical and biological properties of all liposomes and lipoplexes were evaluated, including lipid-DNA interactions, size, morphology, zeta potential, acid-base buffering capability, protection of DNA from DNase I digestion, and cytotoxity. The efficiency of luciferase gene transfection of lipoplexes 1-3 was compared with that of commercial dioleoyl-trimethylammonium propane (DOTAP) and polyethyleneimine (PEI) in 293T cells and HepG2 cells with or without poly(ethylene glycol) PEG stabilizer. The complexation and protection of DNA of liposome 3 was the strongest among the three liposomes. The efficiency of gene transfection of liposomes 1-3 was two-to threefold higher than that of PEI and/or DOTAP in 293T cells. Liposomes 1 and 3 in PEG as stabilizer showed sixfold higher transfection efficiency than that of PEI and/or DOTAP, whereas liposome 2 showed very low transfection efficiency. In HepG2 cells, the transfection efficiency of all the cationic liposomes was much lower than that of DOTAP. In conclusion, lipids 1-3 were efficient and non-toxic gene vectors; the headgroup of cationic lipids and the stabilizer of liposome formulation had an important influence on gene transfection.