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1.
Graphene oxide (GO) has been extensively explored in nanomedicine for its excellent physiochemical, electrical, and optical properties. Here, polyethylene glycol (PEG) and polyethylenimine (PEI) are covalently conjugated to GO via amide bonds, obtaining a physiologically stable dual‐polymer‐functionalized nano‐GO conjugate (NGO‐PEG‐PEI) with ultra‐small size. Compared with free PEI and the GO‐PEI conjugate without PEGylation, NGO‐PEG‐PEI shows superior gene transfection efficiency without serum interference, as well as reduced cytotoxicity. Utilizing the NIR optical absorbance of NGO, the cellular uptake of NGO‐PEG‐PEI is shown to be enhanced under a low power NIR laser irradiation, owing to the mild photothermal heating that increases the cell membrane permeability without significantly damaging cells. As the results, remarkably enhanced plasmid DNA transfection efficiencies induced by the NIR laser are achieved using NGO‐PEG‐PEI as the light‐responsive gene carrier. More importantly, it is shown that our NGO‐PEG‐PEI is able to deliver small interfering RNA (siRNA) into cells under the control of NIR light, resulting in obvious down‐regulation of the target gene, Polo‐like kinase 1 (Plk1), in the presence of laser irradiation. This study is the first to use photothermally enhanced intracellular trafficking of nanocarriers for light‐controllable gene delivery. This work also encourages further explorations of functionalized nano‐GO as a photocontrollable nanovector for combined photothermal and gene therapies.  相似文献   

2.
Polyethylenimine (PEI) is an efficient cationic polymer for gene delivery, but defective in biocompatibility. In this study, we developed two different strategies to shield the positively charged PEI/DNA complexes: PEGylation and lipid coating. The physicochemical properties, cytotoxicity and transfection efficiency of the two gene delivery systems were investigated. Both PEGylation and lipid coating succeeded in reducing the zeta-potential of the complexes. Lipid-coated PEI/DNA complexes (LPD complexes) and PEI/DNA complexes exhibited similar cytotoxicity, whereas PEG-PEI/DNA complexes showed lower cytotoxicity, especially at high N/P ratios. LPD complexes were less efficient in transfection compared to PEG-PEI/DNA complexes. The transfection efficiency was influenced remarkably by cytotoxicity and surface charge of the complexes. Intracellular processes studies revealed that endosomal release might be one of the rate-limiting steps in cell transfection with PEI as a gene delivery carrier.  相似文献   

3.
Wang X  Pei X  Du Y  Li Y 《Nanotechnology》2008,19(37):375102
Non-viral vectors have gained increasing attention in gene therapy because of their safety, but with the shortcoming of low transfection efficiency. We have developed a hybrid material as a novel non-viral vector, which combines the advantages of both biopolymer and clay in a gene delivery system. Quaternized chitosan was intercalated into the interlayers of rectorite to obtain a new polymer/layered silicate nanocomposite. In vitro and in vivo toxicity studies revealed that the nanocomposites were biocompatible and non-toxic. At the nanocomposite:pDNA mass ratio of 8:1, they achieved 100% pDNA adsorption capacity. In vitro cell transfection revealed a transfection efficiency of 32.1% at 96?h as shown by a flow-cytometric study, and the intensive green fluorescence protein (GFP) expression could last for up to 120?h. Furthermore, an in vivo transfection study showed that the most prominent GFP expression was observed in the gastric and duodenum mucosa, and good transfection efficiency was also obtained when injected into the muscle. All the results suggest that quaternized chitosan/rectorite nanocomposite is a novel and potential non-viral gene carrier.  相似文献   

4.
A high‐sensitivity and low‐power theranostic nanosystem that combines with synergistic photothermal therapy and surface‐enhanced Raman scattering (SERS) mapping is constructed by mesoporous silica self‐assembly on the reduced graphene oxide (rGO) nanosheets with nanogap‐aligned gold nanoparticles (AuNPs) encapsulated and arranged inside the nanochannels of the mesoporous silica layer. Rhodamine 6G (R6G) as a Raman reporter is then encapsulated into the nanochannels and anti‐epidermal growth factor receptor (EGFR) is conjugated on the nanocomposite surface, defined as anti‐EGFR‐PEG‐rGO@CPSS‐Au‐R6G, where PEG is polyethylene glycol and CPSS is carbon porous silica nanosheets. SERS spectra results show that rGO@CPSS‐Au‐R6G enhances 5 × 106 magnification of the Raman signals and thus can be applied in the noninvasive cell tracking. Furthermore, it displays high sensitivity (detection limits: 10?8m R6G solution) due to the “hot spots” effects by the arrangements of AuNPs in the nanochannels of mesoporous silica. The highly selective targeting of overexpressing EGFR lung cancer cells (A549) is observed in the anti‐EGFR‐PEG‐rGO@CPSS‐Au‐R6G, in contrast to normal cells (MRC‐5). High photothermal therapy efficiency with a low power density (0.5 W cm?2) of near‐infrared laser can be achieved because of the synergistic effect by conjugated AuNPs and rGO nanosheets. These results demonstrate that the anti‐EGFR‐PEG‐rGO@CPSS‐Au‐R6G is an excellent new theranostic nanosystem with cell targeting, cell tracking, and photothermal therapy capabilities.  相似文献   

5.
In the context of diligent efforts to improve the tumor targeting efficiency of drug carriers, a shape‐persistent polymersome which possess a pH‐tunable membrane as well as folate targeting antennae is reported. The membrane of such polymersomes behaves as gate which undergoes “on” and “off” switches in response to pH stimuli. Thus, polymersomes can effectively prohibit the premature release of chemotherapeutic agents such as doxorubicin in physiological conditions, but promote drug release once they are triggered in the acidified endosomal compartment. Importantly, the folate moieties are installed on the surface of polymersomes as protruding antennae by doping the polymersomes with folate‐terminated block copolymers designed to have longer PEG segments. Thereby, the folate moieties are freed from concealment and steric effects exerted by the dense PEG corona. The cellular uptake of the FA‐antennae polymersomes by tumor cells is significantly enhanced facilitated by the freely accessible folate antennae; however, the normal cells record a low level of cellular uptake due to the stealth property of the PEG corona. Overall, the excellent biocompatibility, controlled permeability, targeted internalization, as well as selective cytotoxicity of such polymersomes set up the basis for properly smart carrier for targeted drug delivery.  相似文献   

6.
While lipoplex (cationic lipid‐nucleic acid complex)‐mediated intracellular delivery is widely adopted in mammalian cell transfection, its transfection efficiency for suspension cells, e.g., lymphatic and hematopoietic cells, is reported at only ≈5% or even lower. Here, efficient and consistent lipoplex‐mediated transfection is demonstrated for hard‐to‐transfect suspension cells via a single‐cell, droplet‐microfluidics approach. In these microdroplets, monodisperse lipoplexes for effective gene delivery are generated via chaotic mixing induced by the serpentine microchannel and co‐confined with single cells. Moreover, the cell membrane permeability increases due to the shear stress exerted on the single cells when they pass through the droplet pinch‐off junction. The transfection efficiency, examined by the delivery of the pcDNA3‐EGFP plasmid, improves from ≈5% to ≈50% for all three tested suspension cell lines, i.e., K562, THP‐1, Jurkat, and with significantly reduced cell‐to‐cell variation, compared to the bulk method. Efficient targeted knockout of the TP53BP1 gene for K562 cells via the CRISPR (clustered regularly interspaced short palindromic repeats)–CAS9 (CRISPR‐associated nuclease 9) mechanism is also achieved using this platform. Lipoplex‐mediated single‐cell transfection via droplet microfluidics is expected to have broad applications in gene therapy and regenerative medicine by providing high transfection efficiency and low cell‐to‐cell variation for hard‐to‐transfect suspension cells.  相似文献   

7.
A majority of the photo‐responsive drug‐delivery systems that are currently being studied require a complicated synthesis method. Here, we prepare a near‐infrared responsive, photothermally controllable, drug‐delivery carrier by a simple mixing and extraction process without the incorporation of toxic chemicals. A blend of doxorubicin (DOX), an anticancer drug, and a phase‐change material (PCM) are loaded onto the mesoporous structure of silica‐coated graphene oxide (GO@MS) to form a waffle‐like structure, which is confirmed by various physicochemical analyses. The cytotoxicity of DOX/PCM‐loaded GO@MS (DOX/PCM‐GO@MS) against HeLa cells is 50 times higher than that of free DOX, and this improved activity can be attributed to the photothermal effectiveness of GO@MS. Additionally, the cytotoxicity and uptake mechanism of the PCM‐based material are analyzed by flow cytometry. Taken together, our results suggest an enormous potential for spatio‐temporal control in photothermally responsive drug‐delivery systems.  相似文献   

8.
Cross‐linked rather than non‐covalently bonded graphitic carbon nitride (g‐C3N4)/reduced graphene oxide (rGO) nanocomposites with tunable band structures have been successfully fabricated by thermal treatment of a mixture of cyanamide and graphene oxide with different weight ratios. The experimental results indicate that compared to pure g‐C3N4, the fabricated CN/rGO nanocomposites show narrowed bandgaps with an increased in the rGO ratio. Furthermore, the band structure of the CN/rGO nanocomposites can be readily tuned by simply controlling the weight ratio of the rGO. It is found that an appropriate rGO ratio in nanocomposite leads to a noticeable positively shifted valence band edge potential, meaning an increased oxidation power. The tunable band structure of the CN/rGO nanocomposites can be ascribed to the formation of C?O?C covalent bonding between the rGO and g‐C3N4 layers, which is experimentally confirmed by Fourier transform infrared (FT‐IR) and X‐ray photoelectron (XPS) data. The resulting nanocomposites are evaluated as photocatalysts by photocatalytic degradation of rhodamine B (RhB) and 4‐nitrophenol under visible light irradiation (λ > 400 nm). The results demonstrate that the photocatalytic activities of the CN/rGO nanocomposites are strongly influenced by rGO ratio. With a rGO ratio of 2.5%, the CN/rGO‐2.5% nanocomposite exhibits the highest photocatalytic efficiency, which is almost 3.0 and 2.7 times that of pure g‐C3N4 toward photocatalytic degradation of RhB and 4‐nitrophenol, respectively. This improved photocatalytic activity could be attributed to the improved visible light utilization, oxidation power, and electron transport property, due to the significantly narrowed bandgap, positively shifted valence band‐edge potential, and enhanced electronic conductivity.  相似文献   

9.
Sensitive and selective detection of nitric oxide (NO) in the human body is crucial since it has the vital roles in the physiological and pathological processes. This study reports a new type of electrochemical NO biosensor based on zinc‐dithiooxamide framework derived porous ZnO nanoparticles and polyterthiophene‐rGO composite. By taking advantage of the synergetic effect between ZnO and poly(TTBA‐rGO) (TTBA = 3′‐(p‐benzoic acid)‐2,2′:5′,2″‐terthiophene, rGO = reduced graphene oxide) nanocomposite layer, the poly(TTBA‐rGO)/ZnO sensor probe displays excellent electrocatalytic activity and explores to detect NO released from normal and cancer cell lines. The ZnO is immobilized on a composite layer of poly(TTBA‐rGO). The highly porous ZnO offers a high electrolyte accessible surface area and high ion–electron transport rates that efficiently catalyze the NO reduction reaction. Amperometry with the modified electrode displays highly sensitive response and wide dynamic range of 0.019–76 × 10?6m with the detection limit of 7.7 ± 0.43 × 10?9m . The sensor probe is demonstrated to detect NO released from living cells by drug stimulation. The proposed sensor provides a powerful platform for the low detection limit that is feasible for real‐time analysis of NO in a biological system.  相似文献   

10.
The intracellular delivery and functionalization of genetic molecules play critical roles in gene‐based theranostics. In particular, the delivery of plasmid DNA (pDNA) with safe nonviral vectors for efficient intracellular gene expression has received increasing attention; however, it still has some limitations. A facile one‐pot method is employed to encapsulate pDNA into zeolitic imidazole framework‐8 (ZIF‐8) and ZIF‐8‐polymer vectors via biomimetic mineralization and coprecipitation. The pDNA molecules are found to be well distributed inside both nanostructures and benefit from their protection against enzymatic degradation. Moreover, through the use of a polyethyleneimine (PEI) 25 kD capping agent, the nanostructures exhibit enhanced loading capacity, better pH responsive release, and stronger binding affinity to pDNA. From in vitro experiments, the cellular uptake and endosomal escape of the protected pDNA are greatly improved with the superior ZIF‐8‐PEI 25 kD vector, leading to successful gene expression with high transfection efficacy, comparable to expensive commercial agents. New cost‐effective avenues to develop metal–organic‐framework‐based nonviral vectors for efficient gene delivery and expression are provided.  相似文献   

11.
生物活性玻璃由于含有钙、磷成分, 因此在基因转染方面具有应用潜能。本研究通过溶胶-凝胶结合静电纺丝技术制备了具有连续介孔或分级纳米孔的微纳米生物活性玻璃纤维(BGF), 并将其用于基因转染。实验结果显示, BGF在无血清培养基中可大量释放Ca2+和PO43-, 这些释放的离子在沉积的同时可装载质粒DNA, 起到基因转染载体的作用, 转染效率与BGF之间具有剂量依赖性。当质粒浓度为1 μg/mL、BGF浓度为1000 μg/mL时, 细胞的转染效率可达对照组(脂质体转染试剂)的50%以上。其转染机理与传统的磷酸钙基因转染类似, 而其较好的离子溶出保障了其使用的稳定性, 有望替代纳米磷酸钙转染体系用于基因传输。  相似文献   

12.
A facile method for PEGylated bioreducible poly(amido amine)s is described by a one-pot Michael-type addition polymerization of N, N′-cystaminebisacrylamide (CBA) with a mixture of 4-amino-1-butanol (ABOL) and mono-tert-butoxycarbonyl (Boc) PEG diamine. By this approach, two Boc-amino-PEGylated p(CBA-ABOL) copolymers were obtained with the PEG/ABOL composition ratio of 1/10 (1a) and 1/6 (2a), respectively. These copolymers were characterized by 1H NMR and gel permeation chromatography. The PEGylated copolymers 1a, and its deprotected analog 1b with a terminal amino group at the PEG chain, were further evaluated as gene delivery vectors. The copolymers 1a and 1b condense DNA into nano-scaled PEGylated polyplexes (< 250 nm) with near neutral (2–5 mV, 1a) or slightly positive (9–13 mV, 1b) surface charge which remain stable in 150 mM buffer solution over 24 h. UnPEGylated polyplexes from p(CBA-ABOL), however, are relatively less stable and increase in size to more than 1 μm. The PEGylated polyplexes showed very low cytotoxicity in MCF-7 and NIH 3T3 cells and induced appreciable transfection efficiencies in the presence of 10% serum, although that are lower than those of p(CBA-ABOL) lacking PEG. The lower transfection efficiency of the PEGylated p(CBA-ABOL) polyplexes is discussed regarding the effect of PEGylation on endosomal escape of the PEGylated polyplexes.  相似文献   

13.
GO and Co(NO3)2 were respectively used as rGO and Co3O4 precursors for preparing magnetically separable Co3O4NPs attached Co3O4NPs@rGO nanocomposites by a straightforward sol–gel technique. To characterize the nanocomposite materials, FESEM, EDX, elemental mapping, XRD, FTIR, Raman spectroscopy, UV–vis, VSM and BET were employed. When exposed to UV rays, the nanocomposite showed extraordinary photocatalytic degradation of MO dye. According to the measurements of photocatalytic activity, the highly efficient photocatalytic efficiency of the nanocomposite could be attributed to preventing electron-hole recombination by highly effective electron transfer between rGO and semiconductor NPs. The nanocomposite succeeded in the efficient degradation of MO dye, even after five photocatalytic cycles.  相似文献   

14.
Sodium carboxymethyl cellulose/reduced graphene oxide (NaCMC/rGO) nanocomposite films were prepared by a simple solution mixing-evaporation method. The NaCMC/rGO nanocomposite films were characterized and compared with sodium carboxymethyl cellulose/graphene oxide (NaCMC/GO) nanocomposite films. The stability of the rGO dispersion, and the structural and mechanical properties of the composite films were investigated by UV–Vis spectrophotometry, X-ray diffraction (XRD), scanning electron microscopy (SEM), Raman spectroscopy, and using a universal testing machine (UTM). The results revealed that CMC and rGO were able to form a homogenous mixture. Compared with pure CMC, the tensile strength and Young's modulus of the CMC/rGO nanocomposite films were considerably enhanced (by 72.52% and 131.79%, respectively) upon incorporation of 2 wt% rGO.  相似文献   

15.
RGD peptides have been incorporated into several gene delivery vehicles to enhance specific interactions of nonviral vehicles with the cell surface. However, there are contradictory results regarding the effect of linear RGD peptides on specific cell surface binding of polyethylene glycol (PEG)-conjugated gene delivery vehicles. This study sought to understand how coupling RGD peptides to PEG vehicles affects cell binding and internalization using a novel four arm PEG backbone. Coupling multiple RGD peptides to the PEG backbone increased the affinity of the vehicle for the cell surface, and that the PEG backbone did not reduce the affinity of RGD peptides for integrin receptors in both kinetic and equilibrium studies. Kinetic studies suggest that cellular internalization of PEG-based vehicles is not regulated by the RGD peptides on the vehicle, but rather by nonspecific interactions with heparan sulfate proteoglycans either alone or in combination with integrins. These results suggest that while increasing the number of RGD peptides per vehicle increases cell binding, but it does not contribute to increased internalization or transfection efficiency.  相似文献   

16.
Novel magnetite-silica nanocomposite particles were prepared using SBA-15 nanoporous silica as template. Magnetite nanoparticles were impregnated into the nanopore array of the silica template through thermal decomposition of iron(III) acetylacetonate, Fe(AcAc)3 at 200 degrees C. These composite particles were characterized using TEM, XRD and SQUID magnetometry. The TEM images showed that the size of composite particles was around 500 nm and the particles retained the nanoporous array of SBA-15. The formation of magnetite nanoparticles was confirmed by the powder XRD study. These composite particles also exhibited ferrimagnetic properties. By coating with short chain polyethyleneimine (PEI), these particles are capable of binding DNA molecules for gene delivery and transfection. With an external magnetic field, the transfection efficiency was shown to have an increase of around 15%. The results indicated that these composite nanoparticles may be further developed as a new tool for nanomagnetic gene transfection.  相似文献   

17.
Co‐delivery of both chemotherapy drugs and siRNA from a single delivery vehicle can have a significant impact on cancer therapy due to the potential for overcoming issues such as drug resistance. However, the inherent chemical differences between charged nucleic acids and hydrophobic drugs have hindered entrapment of both components within a single carrier. While poly(ethylene glycol)‐block‐poly(lactic‐co‐glycolic acid) (PEG–PLGA) copolymers have been used successfully for targeted delivery of chemotherapy drugs, loading of DNA or RNA has been poor. It is demonstrated that significant amounts of DNA can be encapsulated within PLGA‐containing nanoparticles through the use of a new synthetic DNA analog, click nucleic acids (CNAs). First, triblock copolymers of PEG‐CNA‐PLGA are synthesized and then formulated into polymer nanoparticles from oil‐in‐water emulsions. The CNA‐containing particles show high encapsulation of DNA complementary to the CNA sequence, whereas PEG‐PLGA alone shows minimal DNA loading, and non‐complementary DNA strands do not get encapsulated within the PEG‐CNA‐PLGA nanoparticles. Furthermore, the dye pyrene can be successfully co‐loaded with DNA and lastly, a complex, larger DNA sequence that contains an overhang complementary to the CNA can also be encapsulated, demonstrating the potential utility of the CNA‐containing particles as carriers for chemotherapy agents and gene silencers.  相似文献   

18.
Reduced graphene oxide nanomesh (rGONM), as one of the recent structures of graphene with a surprisingly strong near‐infrared (NIR) absorption, is used for achieving ultraefficient photothermal therapy. First, by using TiO2 nanoparticles, graphene oxide nanoplatelets (GONPs) are transformed into GONMs through photocatalytic degradation. Then rGONMs functionalized by polyethylene glycol (PEG), arginine–glycine–aspartic acid (RGD)‐based peptide, and cyanine 7 (Cy7) are utilized for in vivo tumor targeting and fluorescence imaging of human glioblastoma U87MG tumors having ανβ3 integrin receptors, in mouse models. The rGONM‐PEG suspension (1 μg mL?1) exhibits about 4.2‐ and 22.4‐fold higher NIR absorption at 808 nm than rGONP‐PEG and graphene oxide (GO) with lateral dimensions of ≈60 nm and ≈2 μm. In vivo fluorescence imaging demonstrates high selective tumor uptake of rGONM‐PEG‐Cy7‐RGD in mice bearing U87MG cells. The excellent NIR absorbance and tumor targeting of rGONM‐PEG‐Cy7‐RGD results in an ultraefficient photothermal therapy (100% tumor elimination 48 h after intravenous injection of an ultralow concentration (10 μg mL?1) of rGONM‐PEG‐Cy7‐RGD followed by irradiation with an ultralow laser power (0.1 W cm?2) for 7 min), whereas the corresponding rGO‐ and rGONP‐based composites do not present remarkable treatments under the same conditions. All the mice treated by rGONM‐PEG‐Cy7‐RGD survived over 100 days, whereas the mice treated by other usual rGO‐based composites were dead before 38 days. The results introduce rGONM as one of the most promising nanomaterials in developing highly desired ultraefficient photothermal therapy.  相似文献   

19.
Nano‐sized in vivo active targeting drug delivery systems have been developed to a high anti‐tumor efficacy strategy against certain cancer‐cells‐specific. Graphene based nanocarriers with unique physical and chemical properties have shown significant potentials in this aspect. Here, octreotide (OCT), an efficient biotarget molecule, is conjugated to PEGylated nanographene oxide (NGO) drug carriers for the first time. The obtained NGO‐PEG‐OCT complex shows low toxicity and excellent stability in vivo and is able to achieve somatostatin receptor‐mediated tumor‐specific targeting delivery. Owing to the high loading efficiency and accurate targeting delivery of anti‐cancer drug doxorubicin (DOX), our DOX loaded NGO‐PEG‐OCT complex offers a remarkably improved cancer‐cell‐specific cellular uptake, chemo‐cytotoxicity, and decreased systemic toxicity compared to free DOX or NGO‐PEG. More importantly, due to its strong near‐infrared absorption, the NGO‐PEG‐OCT complex further enhances efficient photothermal ablation of tumors, delivering combined chemo and photothermal therapeutic effect against cancer cells.  相似文献   

20.
Gene therapy has been considered a strategy for delivery of therapeutic nucleic acids to a specific site. Calcium phosphates are one gene delivery vector group of interest. However, low transfection efficiency has limited the use of calcium phosphate in gene delivery applications. Present work aims at studying the fabrication of strontium substituted calcium phosphate nanoparticles with improved gene delivery related properties. Strontium substituted calcium phosphate was prepared using a simple sol gel method. X-ray diffraction analysis, Fourier transform infrared spectroscopy, transmission electron microscopy, specific surface area analysis, zeta potential measurement and ion release evaluation were used to characterize the samples. This characterization showed strontium and carbonate co-substituted calcium phosphate which resulted in nano size particles with low crystallinity, high specific surface area, positive surface charge, and a high dissolution rate. These improved properties could increase the DNA concentration on the vector as well as the endosomal escape of the complex that leads to higher transfection efficiency of this novel gene delivery vector.  相似文献   

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