Polymers
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2018
·Vol. 10
·期 10
·1060
·
IF 2.935
Miao-Miao Xun,Zheng Huang,Ya-Ping Xiao,Yan-Hong Liu,Ji Zhang,Ju-Hui Zhang,Xiao-Qi Yu
Rapid enzymatic degradation and fragmentation during DNA administration can result in limited gene expression, and consequently, poor efficacy. It is necessary to use novel vectors for DNA delivery. Herein, we aimed to design useful carriers for enhancing transfection efficiency (TE). These lipopolymers were prepared through Michael addition reactions from low-molecular-weight (LMW) polyethyleneimine (PEI) and linkers with three kinds of steroids. Agarose gel electrophoresis assay results displayed that the three lipopolymers could condense plasmid DNA well, and the formed polyplexes had appropriate sizes around 200–300 nm, and zeta potentials of about +25–40 mV. The results of in vitro experiments using HeLa, HEK293, and MCF-7 cells showed that these lipopolymers present higher TE than 25-kDa PEI, both in the absence and presence of 10% serum. Flow cytometry and confocal microscopy studies also demonstrated that these lipopolymer/DNA complexes present higher cellular uptake and intracellular distribution. The measurement of critical micelle concentration (CMC) revealed that these lipopolymers could form micelles, which are suited for drug delivery. All results suggest that the three materials may serve as hopeful candidates for gene and drug delivery in future in vivo applications.Keywords:gene vectors;Michael addition;lipopolymers;steroids
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TALANTA
·
2018
·
IF 4.244
Junqing He,Junyan Wang,Nan Zhang,Luyao Shen,Linlin Wang,Xiao Xiao,Yan Wang,Tao Bing,Xiangjun Liu,Songqing Li,Dihua Shangguan
Ovarian cancer is regarded as the most lethal gynecologic malignancy with poor prognosis and high mortality rate. Drug-resistance was thought to be the main obstacle to improving overall survival rate of ovarian cancer. New ligands for drug-resistant ovarian cancer cells have potential for the development of diagnosis and therapy of ovarian cancer. In present work, we reported two aptamers , HF3–58 and HA5–68 generated by cell-SELEX, against a paclitaxel-resistant ovarian cancer cell line (A2780T). Both two aptamers exhibited high selectivity and strong affinity to target cells with low nanomolar dissociation constants . The binding of aptamers to target cells was independent of divalent ions, and was tolerant of incubation temperature and nucleases in serum. Molecular targets of the two aptamers were preliminarily demonstrated to be two different glycoproteins on cell surface of A2780T cells. Owing to the structure stability and high resistance to nuclease, these two aptamers had good performance in the detection of drug-resistant ovarian cancer cells in human serum.
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BIOMATERIALS
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2012
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IF 7.404
Yiyan He,Gang Cheng,Li Xie,Yu Nie,Bin He,Zhongwei Gu
The natural anionic polysaccharide hyaluronic acid (HA) was modified by introducing reduction-sensitive disulfide bond between the carboxyl groups and the backbone of HA (HA-SS-COOH). HA-SS-COOH and its corresponding unmodified stable analog HA were used to shield DNA/PEI polyplexes (DP) to form ternary complexes (DPS and DPH complexes). The shielding/deshielding effect was tested along with size, zeta potential , cell viability and transfection. Both DPS and DPH complexes showed increase in size, decrease in zeta potential and low cytotoxicity in physiological conditions due to the anionic shielding. In the reductive environment, only HA-SS-COOH coated ternary complexes (DPS) demonstrated the size increase and recovered high positive zeta potential. DPS complexes showed an up to 14-fold higher transfection than the stable coated one, indicating the efficiency of the reduction-responsive deshielding design. Moreover, the presence of extra free HA inhibited the transfection of DPS on HepG2 and B16F10 cells with HA receptor expression, while displaying no effect on non-targeted NIH3T3 cells. More rapid cellular association of DPS with HepG2 was observed, thus confirming the targeting reservation of disulfide bond modified HA. Intratumoral injection of DPS complexes resulted in much higher accumulation and luciferase expression in the tumor bearing C57BL/6 mice. Both in vitro and in vivo results demonstrated the successful combination of deshielding and target functions in HA derivatives for gene delivery.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2014
·
IF 3.84
Qionghua Zhu,Feng Gao,Yizhen Yang,Bin Zhang,Wei Wang,Zhengshui Hu,Qingxiang Wang
A highly sensitive impedimetric DNA biosensor based on rod-like bismuth sulfide nano (rBi 2 S 3 ) and polyaniline (PANI) nanocomposite film modified ionic liquid-carbon paste electrode (IL-CPE) has been developed. The rBi 2 S 3 particles were first dropt on IL-CPE that was constituted with 1-butyl-3-methylimidazolium hexafluorophosphate (BMIMPF 6 ) and graphite powder, and then the PANI polymer was deposited on the surface of rBi 2 S 3 through potentiodynamic polymerization using aniline as the source, forming the PANI capped Bi 2 S 3 nanocomposite. The morphologies and electrocatalytic property of the composite were characterized by scanning electron microscopy (SEM) and electrochemical techniques including electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV), respectively. The integrated advantages such as the large surface area, excellent electronic conductivity, easy to fabrication, and good biocompatibility make the electrode favorable for DNA immobilization and hybridization detection. It was found that remarkable increase of electrochemical impedance was observed after the probe DNA modified electrode was hybridized with the target DNA. The dynamic detection range was determined to be from 1.0 × 10 −15 to 1.0 × 10 −11 M with a detection limit of 4.37 × 10 −16 M. The DNA biosensor also displayed good regeneration ability, reproducibility and stability.
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ACS Applied Materials & Interfaces
·
2015
·
IF 6.723
Yanling Hu,Lei Zhang,Ying Zhang,Bin Wang,Yawei Wang,Quli Fan,Wei Huang,Lianhui Wang
MicroRNAs (miRNAs), a class of small, endogenous, noncoding RNA molecules, can serve as biomarkers for potential applications in cancer diagnosis, prognosis, and prediction due to its abnormal expression. As a result, a novel label-free biosensor with nanometer scale was prepared and employed in the detection of trace oligonucleotides based on the localized surface plasmon resonance (LSPR). The dielectric constant on the surface of DNA modified gold nanoparticle would change when probe single-strand DNA hybridized with target oligonucleotides, which resulted in the notable red shift of scattering peak position. The biosensor with excellent selectivity can be used in a real-time monitoring hybridization process. Notably, this method provided label-free detection of DNA and miRNA at single nanoparticle level with limit of detection up to 3 nM. Due to the advantages of LSPR scattering spectra, single nanoparticle biosensor can be designed for trace cancer-relevant miRNAs detection in the future.
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ANALYTICA CHIMICA ACTA
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2014
·
IF 4.517
Lichun Zheng,Xiuhui Liu,Min Zhou,Yongjun Ma,Guofan Wu,Xiaoquan Lu
We presented a new strategy for ultrasensitive detection of DNA sequences based on the novel detection probe which was labeled with Ag + using metallothionein (MT) as a bridge. The assay relied on a sandwich-type DNA hybridization in which the DNA targets were first hybridized to the captured oligonucleotide probes immobilized on Fe 3 O 4 @Au composite magnetic nanoparticles (MNPs), and then the Ag + -modified detection probes were used to monitor the presence of the specific DNA targets. After being anchored on the hybrids, Ag + was released down through acidic treatment and sensitively determined by a coupling flow injection–chemiluminescent reaction system (Ag + –Mn 2+ –K 2 S 2 O 8 –H 3 PO 4 –luminol) (FI–CL). The experiment results showed that the CL intensities increased linearly with the concentrations of DNA targets in the range from 10 to 500 pmol L −1 with a detection limit of 3.3 pmol L −1 . The high sensitivity in this work may be ascribed to the high molar ratio of Ag + –MT, the sensitive determination of Ag + by the coupling FI–CL reaction system and the perfect magnetic separation based on Fe 3 O 4 @Au composite MNPs. Moreover, the proposed strategy exhibited excellent selectivity against the mismatched DNA sequences and could be applied to real samples analysis.
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MICROCHIMICA ACTA
·
2013
·Vol. 181
·期 1
·155-162
·
IF 3.434
Wang Liheng,Liao Xiaolei,Ding Yingtao,Gao Fei,Wang Qingxiang
Abstract We report on an electrochemical DNA biosensor consisting of a glassy carbon electrode modified with a film of electropolymerized Eriochrome Black T (pEBT) that serves as a functional platform for the immobilization of probe DNA. pEBT was deposited via cyclic voltammetry, and the amino-modified DNA capture probe was covalently linked to the surface via a sulfanilamide coupling reaction. The single step of the assembly process was monitored by atomic force microscopy and electrochemistry. The surface density of DNA probe on the biosensor interface was calculated to be 1.7 × 10 −10 mol cm −2 using methylene blue as an electroactive probe. Hybridization experiments showed the peak currents of methylene blue to decrease with increasing concentration of complementary sequence in the range from 5.0 f. to 5.0 pM. The detection limit is as low as 0.11 fM. Selectivity studies showed that the biosensor can discriminate a fully complementary sequence from a single-base mismatch, three-base mismatch, and a fully non-complementary sequence. The biosensor displays good stability and can be regenerated due to the beneficial effects of electropolymerization and covalent immobilization of probe DNA.
A novel DNA biosensor was fabricated through a facile sulfamide coupling reaction between probe DNA and eriochrome black T that electropolymerized on a glassy carbon electrode
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BIOSENSORS & BIOELECTRONICS
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2013
·
IF 5.437
Dan Zhao,Jiaotian Li,Tianming Yang,Zhike He
A “turn off–on” mode has been established by using the interaction between platinum anticancer drugs and DNA as input signal and the fluorescence reversible change of quantum dots (QDs) as output signal. The QDs fluorescence can be quenched by platinum anticancer drugs via photo-induced electron transfer process, rendering the system into “turn off” status, and the system can then be “turned on” when fluorescence is restored due to covalent conjugation between DNA and platinum anticancer drugs. This dual-directional fluorescence change realized the detection of cisplatin and DNA, overcoming the selectivity problem commonly existed in the traditional mono-directional fluorescence detection mode. The reversible fluorescent “turn off-on” mode has been further employed to study the interactions between DNA and different platinum anticancer drugs (cisplatin, oxaliplatin and carboplatin). Furthermore, the impacts of different types of DNAs (different in base sequence , chain length and ssDNA/dsDNA) on the mode are also explored. This simple, fast and convenient spectroscopic method owns promising applications in the study on interaction between medical molecules and DNA, and in biochemical detections.
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BIOSENSORS & BIOELECTRONICS
·
2013
·
IF 5.437
Shuang Dai,Qingwang Xue,Jing Zhu,Yongshun Ding,Wei Jiang,Lei Wang
An ultrasensitive fluorescence method for determination of protein is developed based on hybridization chain reaction (HCR). In this assay, the streptavidin-magnetic nanobeads were conjugated to biotinylated initiators and biotinylated anti-IgG. In the presence of human IgG, the magnetic nanobeads were fixed on the substrate and the carried initiators propagated the chain reaction of hybridization to form the nicked polymers. Because the nanobead probe carries with a large number of oligonucleotides per protein binding event, there is obvious amplification in the nicked polymers. Then, numerous SYBR Green I molecules were intercalated into the grooves of the long dsDNA polymers, generating a substantially apparent increase in the corresponding fluorescence intensity . With HCR amplification and magenetic nanobead to preamplify the fluorescence signal and reduce the background signal, the detection limit of this assay was 14 aM. Compared with the reported protein detection methods, our method exhibited ultrahigh sensitivity. In addition, the proposed method possessed excellent selectivity and low matrix effect . What is more, the assay was also studied for clinical application in human serum with a satisfactory and reliable result.
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MACROMOLECULAR CHEMISTRY AND PHYSICS
·
2013
·Vol. 214
·期 20
·2266-2275
·
IF 2.386
Jixian Liu,Zhen Liu,Xiong Luo,Xidan Zong,Jingquan Liu
The preparation of biodegradable and positively charged graphene–polymer nanocomposite for DNA uptake and release is presented. A pyrene-terminated poly(2-(dimethylamino) ethyl acrylate) is firstly synthesized via reversible addition–fragmentation chain transfer (RAFT) polymerization, followed by the attachment onto the graphene surface via the π–π stacking interaction. The positively charged polymer brushes benefit the uptake of the negatively charged DNA fragment via static interaction and the built-in disulfide linkage between the polymer chain and graphene surface allows the biodegradable release of DNA in the presence of reducing agents. X-ray photoelectron spectroscopy manifests the binding of DNA molecules to the polymer chains and agarose gel electrophoresis confirms the release of DNA.
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MACROMOLECULAR CHEMISTRY AND PHYSICS
·
2015
·Vol. 216
·期 14
·1507-1515
·
IF 2.616
Yunna Kong,Wenyu Li,Qianying Mao,Youxiang Wang
Supramolecular assembly provides a new strategy for designing various gene carriers for deoxyribonucleic acid (DNA) delivery. In this work, supramolecular pseudo-block conjugates based on star-shaped polycation are developed as novel nonviral gene vector. First, cyclodextrin (CD) is selected as core and grafted with low molecular weight polyethylenimine (PEI 1800 ) as arms to obtain high molecular weight star-shaped polycation CD-PEI 8 (SCP). Then, azobenzene (Az) modified poly(ethylene glycol) is synthesized and named as azobenzene modified poly(ethylene glycol) (Az-PEG). Subsequently, the supramolecular pseudo-block conjugates are prepared via host–guest interaction between CD and Az. The result suggests that SCP/Az-PEG condenses DNA into spherical nanoparticles with sizes of around 150 nm. The in vitro experiment shows that the cell viability of DNA/SCP/Az-PEG polyplexes is significantly improved compared with that of PEI 25k /DNA polyplexes. The PEGylation does not decrease the cellular uptake efficiency and transfection efficiency. Furthermore, the cellular trafficking shows that PEGylation does not significantly influence nuclear entry. The star-shaped architecture of polycation plays an important role in gene delivery.
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JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY
·
2015
·
IF 2.96
Jie-Hua Shi,Ting-Ting Liu,Min Jiang,Jun Chen,Qi Wang
The binding interaction of gefitinib with calf thymus DNA (ct-DNA) under the simulated physiological pH condition was studied employing UV absorption, fluorescence, circular dichroism (CD), viscosity measurement and molecular docking methods. The experimental results revealed that gefitinib preferred to bind to the minor groove of ct-DNA with the binding constant ( K b ) of 1.29 × 10 4 L mol −1 at 298 K. Base on the signs and magnitudes of the enthalpy change (Δ H 0 = −60.4 kJ mol −1 ) and entropy change (Δ S 0 = −124.7 J mol −1 K −1 ) in the binding process and the results of molecular docking, it can be concluded that the main interaction forces between gefitinib and ct-DNA in the binding process were van der Waals force and hydrogen bonding interaction. The results of CD experiments revealed that gefitinib did not disturb native B-conformation of ct-DNA. And, the significant change in the conformation of gefitinib in gefitinib–ct-DNA complex was observed from the molecular docking results and the change was close relation with the structure of B-DNA fragments, indicating that the flexibility of gefitinib molecule also plays an important role in the formation of the stable gefitinib–ct-DNA complex.
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