RESUMO
Near-infrared (NIR) persistent luminescence nanoparticles (PLNPs) with high brightness, small sizes, good hydro-dispersivity, and intrinsic surface-functional groups are desirable in biological applications. In this work, Cr3+-doped zinc gallogermanates Zn1+xGa2-2xGexO4:Cr (ZGGC) PLNPs were hydrothermally synthesized via 3-aminopropyltriethoxysilane (APTES) as an additive, or APTES and cetyltrimethylammonium bromide (CTAB) as two co-additives. Addition of APTES not only dramatically enhances the 696 nm NIR luminescence intensity, but also obviously decreases the particle size and introduces amino groups. In particular, thex= 0.1 series ZGGC (ZGGC0.1) with the addition of n moles equivalent APTES (ZGGC0.1-nA) had smaller particle sizes than thex= 0.2 counterpart (ZGGC0.2-nA). The NIR afterglow intensities increased with the APTES introduction. The ZGGC0.2-2.5A sample (also named as ZGGC, Si, -NH2) exhibited maximum luminescence intensities both in solid and aqueous states. With APTES, Si atom is doped and -NH2groups are modified, the trap depth and density become larger, and the afterglow intensities and decay time are significantly enhanced. More notably, co-addition of CTAB (ZGGC0.2-2.5A-C) (also named as ZGGC, Si, -NH2') further enhances hydro-dispersivity and luminescence intensity, decreases particle sizes, and results in more prominent amino groups. The trap density is drastically higher than that without CTAB (i.e. ZGGC0.2-2.5A). Change of Cr3+microenvironment in the crystal and more defects introduction contribute to the enhanced brightness. As expected, the ZGGC,Si,-NH2' PLNPs possess excellent biocompatibility, deep tissue penetration and distinguished bioimaging properties, and rechargeability with orange LED light. The ZGGC,Si,-NH2' PLNPs should provide to be an excellent nanomaterial for various functionalization and bioimaging applications.
Assuntos
Luminescência , Nanopartículas , Cetrimônio , Nanopartículas/química , Tamanho da PartículaRESUMO
The Systematic Evolution of Ligands by EXponential enrichment (SELEX) is conventionally an effective method to identify aptamers, which are oligonucleotide sequences with desired properties to recognize targets specifically and sensitively. However, there are some inherent limitations, e.g., the loss of potential high-affinity sequences during biased iterative PCR enrichment processes and the limited structural diversity of the initial library, which seriously restrict their real-world applications. To overcome these limitations, the in silico base mutagenesis post-SELEX strategy based on the low Gibbs free energy (ΔG) and genetic algorithm was developed for the optimization of the interferon-gamma aptamer (B1-4). In the process of evolution, new sequences were created and the aptamer candidates with low ΔG values and advanced structures were produced. After five rounds of selection, systematic studies revealed that the affinity of the newly developed evolutionary aptamer (M5-5) was roughly 10-fold higher than that of the parent aptamer (B1-4), and an aptasensor detection system with a limit-of-detection (LOD) value of 3.17 nM was established based on the evolutionary aptamer. The proposed approach provided an efficient strategy to improve the aptamer with low energy and a high binding ability, and the good analytical utility thereof.
Assuntos
Aptâmeros de Nucleotídeos , Técnica de Seleção de Aptâmeros , Aptâmeros de Nucleotídeos/química , Interferon gama/genética , Ligantes , Mutagênese , Técnica de Seleção de Aptâmeros/métodosRESUMO
A sensitive and turn-on fluorescence nanoprobe based on core-shell Ag@Au nanoparticles (Ag@AuNPs) as a fluorescence receptor and red emissive graphene quantum dots (GQDs) as a donor was fabricated. They were conjugated together through π-π stacking between the GQDs and single-strand DNA modified at the Ag@AuNPs surface. The absorption spectrum of the receptor significantly overlapped with the donor emission spectrum, leading to a strong Förster resonance energy transfer (FRET) and thus a dramatic quenching. The sensing mechanism relies on fluorescence recovery following DNA cleavage by â¢OH produced from Fenton-like reaction between the peroxidase-like Ag nanocore and H2O2. The red emissive feature (Ex/Em, 520 nm/560 nm) provides low background in physiological samples. The â¢OH production, great spectrum overlapping, and red emission together contributes to good sensitivity and living cell imaging capability. The fluorescence assay (intensity at 560 nm) achieves a low detection limit of 0.49 µM H2O2 and a wide linear range from 5 to 200 µM, superior to most of the reported fluorescent probes. The RSD value for 100 µM H2O2 was 1.4%. The nanoprobe exhibits excellent anti-interferences and shows low cytotoxicity. The recovery of 100 µM standard H2O2 in a cancer cell lysate was 85.8%. Most satisfactorily, it can realize monitoring and imaging H2O2 in living cells. This study not only presents a sensitive H2O2 probe but also provides a platform for detecting other types of reactive oxygen species.
Assuntos
Corantes Fluorescentes/uso terapêutico , Ouro/química , Grafite/química , Peróxido de Hidrogênio/química , Nanopartículas Metálicas/química , Pontos Quânticos/química , Prata/química , HumanosRESUMO
An ultrasensitive enzyme-free electrochemical sandwich DNA biosensor is described for the detection of ssDNA oligonucleotides. A DNA sequence derived from the genom of Helicobacter pylori was selected as a model target DNA. The DNA assay was realized through catching target DNA on capture DNA immobilized gold electrode; then labeling the target DNA with reporter DNA (rpDNA) and initiator DNA (iDNA) co-modified gold nanoparticles (AuNPs). The high density of iDNAs serves as one of the amplification strategies. The iDNA triggers hybridization chain reaction (HCR) between two hairpins. This leads to the formation of a long dsDNA concatamer strand and represents one amplification strategy. The electrochemical probe [Ru(NH3)5L]2+, where L stands for 3-(2-phenanthren-9-ylvinyl)pyridine, intercalated into dsDNA chain. Multiple probe molecules intercalate into one dsDNA chain, serving as one amplification strategy. The electrode was subjected to differential pulse voltammetry for signal acquisition, and the oxidation peak current at -0.28 V was recorded. On each AuNP, 240 iDNA and 25 rpDNA molecules were immobilized. Successful execution of HCR at the DNA-modified AuNPs was confirmed by gel electrophoresis and hydrodynamic diameter measurements. Introduction of HCR significantly enhances the DNA detection signal intensity. The assay has two linear ranges of different slopes, one from 0.01 fM to 0.5 fM; and one from 1 fM to 100 fM. The detection limit is as low as 0.68 aM. Single mismatch DNA can be differentiated from the fully complementary DNA. Conceivably, this highly sensitive and selective assay provides a general method for detection of various kinds of DNA. Graphical abstractSchematic representation of the detection and the amplification principles of the electrochemical sandwich DNA assay. Purple curl: Captured DNA; Green curl: Reporter DNA; Orange curl: HCR initiator DNA; Yellow solid-circle: Gold nanoparticle; H1 and H2: Two hairpin DNA; [Ru(NH3)5L]2+: Signal probe.
Assuntos
Técnicas Biossensoriais , DNA Bacteriano/análise , Técnicas Eletroquímicas , Ouro/química , Helicobacter pylori/química , Nanopartículas Metálicas/química , Hibridização de Ácido Nucleico , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
Hairpin DNA (hpDNA) as a novel biobarcode was conjugated with gold nanoparticles (AuNPs) and a reporter DNA (rpDNA) to form hpDNA/AuNP/rpDNA nanoparticles for the detection of an oligonucleotide sequence associated with Helicobacter pylori as a model target. The rpDNA is complementary to about a half-portion of the target DNA sequence (tDNA). A capture DNA probe (cpDNA), complementary to the other half of the tDNA, was immobilized on the surface of a gold electrode. In the presence of tDNA, a sandwich structure of (hpDNA/AuNP/rpDNA)/tDNA/cpDNA was formed on the electrode surface. The differential pulse voltammetry (DPV) detection was based on [Ru(NH3)5(3-(2-phenanthren-9-yl-vinyl)-pyridine)](2+), an electroactive complex that binds to the sandwich structure by its intercalation with the hpDNA and the double-stranded DNA (dsDNA) of the sandwich structure. The several factors--high density of biobarcode hpDNA on the surface of AuNPs, multiple electroactive complex molecules intercalated with each hpDNA and dsDNA molecule, and the intercalation binding mode of the electroactive complex with the DNA sandwich structure--contribute to the DNA sensor with highly selective and sensitive sensing properties. The DNA sensor exhibited a detection limit of 1 × 10(-15) M (i.e., 1 fM), the DNA levels in physiological samples, with linearity down to 2 × 10(-15) M. It can differentiate even one single mismatched DNA from the complementary tDNA. This novel biobarcode-based DNA sensing approach should provide a general platform for development of direct, simple, repetitive, sensitive, and selective DNA sensors for various important applications in analytical, environmental, and clinical chemistry.
Assuntos
Sondas de DNA/química , DNA Bacteriano/análise , Técnicas Eletroquímicas/instrumentação , Ouro/química , Helicobacter pylori/isolamento & purificação , Nanopartículas Metálicas/química , Complexos de Coordenação/química , DNA Bacteriano/isolamento & purificação , Técnicas Eletroquímicas/métodos , Eletrodos , Desenho de Equipamento , Infecções por Helicobacter/diagnóstico , Infecções por Helicobacter/microbiologia , Humanos , Ácidos Nucleicos Imobilizados/química , Limite de Detecção , Rutênio/químicaRESUMO
OBJECTIVE: To establish an HPLC method for characteristic chromatograms analysis and simultaneous determination of six marker components in Xinnaojian Capsules from different manufactories. METHODS: Using HPLC, an Agilent C18 column (100 mm x 4.6 mm, 2.7 µm) was adopted with acetonitrile-0.05% phosphoric acid as the mobile phase in a gradient elution mode, the flow ratewas 0.4 mL/min, the detection wavelength was 280 nm, and the column temperature was 40 degrees C. RESULTS: Totally eleven common peaks were recognized with epigallocatechin-3-gallate as the reference peak. There were good similarities between the standard characteristic chromatogram and each characteristic chromatogram of the 26 samples with their similarities over 0.99. The six marker components (gallic acid, gallocatechin, caffeine, epigallocatechin, epigallocatechin-3-gallate and epicatechin gallate) were separated well. Good correlation coefficients were found (r > 0.9990) and the average recovery rates ranged from 95.29% to 102.3%. CONCLUSION: The established method has high sensitivity and specificity, and can be used for the quality control of Xinnaojian Capsules.
Assuntos
Medicamentos de Ervas Chinesas/química , Controle de Qualidade , Cafeína/análise , Cápsulas , Catequina/análogos & derivados , Catequina/análise , Cromatografia Líquida de Alta Pressão , Ácido Gálico/análiseRESUMO
A planar boron-doped diamond (BDD) electrode was treated with KOH and functionalized with 3-aminopropyltriethoxysilane (APTES) to serve as a biosensing platform for biomolecule immobilization with glucose oxidase (GOx) as a test model. The free amino groups of GOx and APTES were cross-linked by glutaraldehyde (X), a bifunctional chemical to form a stable enzyme layer (GOx-X-APTES) on BDD. Micrographs obtained by scanning electron microscopy revealed that a mesoporous structure uniformly covered the BDD surface. Cyclic voltammetry of GOx immobilized showed a pair of well-defined redox peaks in neutral phosphate buffer solution, corresponding to the direct electron transfer of GOx. The apparent heterogeneous electron transfer rate constant of the immobilized GOx was estimated to be 8.85 ± 0.47 s(-1), considerably higher than the literature reported values. The determination of glucose was carried out by amperometry at -0.40 V, and the developed biosensor showed good reproducibility and stability with a detection limit of 20 µM. Both ascorbic and uric acids at normal physiological conditions did not provoke any signals. The dynamic range of glucose detection was further extended by covering the enzyme electrode with a thin Nafion layer. The Nafion/GOx-X-APTES/BDD biosensor showed excellent stability, a detection limit of 30 µM, a linear range between 35 µM and 8 mM, and a dynamic range up to 14 mM. Such analytical performances were compared favorably with other complicated sensing schemes using nanomaterials, redox polymers, and nanowires. The APTES-functionalized BDD could be easily extended to immobilize other redox enzymes or proteins of interests.
Assuntos
Boro/química , Diamante/química , Transporte de Elétrons , Enzimas Imobilizadas/química , Glucose Oxidase/química , Técnicas Biossensoriais , Eletrodos , Indicadores e Reagentes , Oxigênio/químicaRESUMO
A thin highly reduced graphene oxide (rGO) film was self-assembled at the dimethyl formamide (DMF)-air interface through evaporation-induced water-assisted thin film formation at the pentane-DMF interface, followed by complete evaporation of pentane. The thin film was transferred onto various solid substrates for film characterization and electrochemical sensing. UV-visible spectrometry, scanning electron microscopy (SEM), atomic force microscopy (AFM) and electrochemistry techniques were used to characterize the film. An rGO film showing 82.8% of the transmittance at 550 nm corresponds to a few layers of rGO nanosheets. The rGO nanosheets cross-stack with each other, lying approximately in the plane of the film. An rGO film collected on a glassy carbon (GC) electrode exhibited improved electrical conductivity compared to GC, with the electrode charge-transfer resistance (Rct) reduced from 31 Ω to 22 Ω. The as-formed rGO/GC electrode was mechanically very stable, exhibiting significantly enhanced electrocatalytic activity to H(2)O(2) and dopamine. Multiple layers of the rGO films on the GC electrode showed even stronger electrocatalytic activity to dopamine than that of the single rGO film layer. The controllable formation of a stable rGO film on various solid substrates has potential applications for nanoelectronics and sensors/biosensors.
Assuntos
Grafite/química , Óxidos/química , Técnicas Biossensoriais/métodos , Catálise , Eletroquímica/métodos , Eletrodos , Peróxido de Hidrogênio/química , Microscopia de Força Atômica/métodos , Microscopia Eletrônica de Varredura/métodos , Oxirredução , Água/químicaRESUMO
Clustered regularly interspaced short palindromic repeat (CRISPR) system has been explored as an efficient tool for nucleic acid diagnostics. However, it normally needs instrumentation or produces turn-off signals. Herein, a bulged Y-shape DNA (Y-DNA) nanoassembly was designed and synthesized as a novel turn-on probe. A CRISPR/Cas12a and Y-DNA probe mediated colorimetric assay (named as CYMCOA) strategy was developed for visual detection of pathogen DNA. Upon activating Cas12a with pathogen DNA, the Y-DNA bulge is catalytically trans-cleaved, releasing the G-quadruplex sequence embedded in the Y-DNA nanoassembly as a peroxidase-like DNAzyme. Visible signals with chromogen substrates are thus produced. The CYMCOA strategy was combined with recombinase polymerase amplification (RPA), an isothermal amplification technique, in detecting Helicobacter pylori (Hp) bacteria and SARS-CoV-2 N plasmids as two model pathogens. The bioassay has very excellent detection sensitivity and specificity, owing to the triple cascade amplification reactions and the very low mismatch tolerance. The lower limit of detection values were 0.16 cfuâ mL-1, 1.5 copiesâ µL-1, and 0.17 copiesâ µL-1 for Hp bacteria, Hp plasmids, and SARS-CoV-2 N plasmids respectively. The detection is fast and accurate. The colorimetric bioassay strategy provides to be a simple, accurate, fast and instrumentation-free platform for nucleic acids detections in various settings, including crude and emergent situations.
Assuntos
Sistemas CRISPR-Cas , Colorimetria , Técnicas de Amplificação de Ácido Nucleico , SARS-CoV-2 , Colorimetria/métodos , Sistemas CRISPR-Cas/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Helicobacter pylori/genética , Helicobacter pylori/isolamento & purificação , DNA Bacteriano/genética , DNA Bacteriano/análise , DNA Viral/genética , DNA Viral/análise , Limite de Detecção , Humanos , Técnicas Biossensoriais/métodos , Nanoestruturas/química , Sondas de DNA/química , Sondas de DNA/genética , Proteínas Associadas a CRISPR/genética , Proteínas de Bactérias/genética , EndodesoxirribonucleasesRESUMO
Immobilization of proteolytic enzymes onto nanocarriers is effective to improve drug diffusion in tumors through degrading the dense extracellular matrix (ECM). Herein, immobilization and release behaviors of hyaluronidase, bromelain, and collagenase (Coll) on mesoporous silica nanoparticles (MSNs) were explored. A series of cationic MSNs (CMSNs) with large and adjustable pore sizes were synthesized, and investigated together with two anionic MSNs of different pore sizes. CMSNs4.0 exhibited the highest enzyme loading capacity for hyaluronidase and bromelain, and CMSNs4.5 was the best for Coll. High electrostatic interaction, matched pore size, and large pore volume and surface area favor the immobilization. Changes of the enzyme conformations and surface charges with pH, existence of a space around the immobilized enzymes, and the depth of the pore structures, affect the release ratio and tunability. The optimal CMSNs-enzyme complexes exhibited deep and homogeneous penetration into pancreatic tumors, a tumor model with the densest ECM, with CMSNs4.5-Coll as the best. Upon loading with doxorubicin (DOX), the CMSNs-enzyme complexes induced high anti-tumor efficiencies. Conceivably, the DOX/CMSNs4.5-NH2-Coll nanodrug exhibited the most effective tumor therapy, with a tumor growth inhibition ratio of 86.1 %. The study provides excellent nanocarrier-enzyme complexes, and offers instructive theories for enhanced tumor penetration and therapy.
Assuntos
Doxorrubicina , Enzimas Imobilizadas , Nanopartículas , Dióxido de Silício , Dióxido de Silício/química , Enzimas Imobilizadas/química , Nanopartículas/química , Porosidade , Doxorrubicina/química , Doxorrubicina/farmacologia , Animais , Humanos , Camundongos , Portadores de Fármacos/química , Linhagem Celular Tumoral , Hialuronoglucosaminidase/química , Hialuronoglucosaminidase/metabolismo , Liberação Controlada de Fármacos , Colagenases/metabolismo , Colagenases/química , Bromelaínas/química , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/patologiaRESUMO
Gastric cancer is one of the most common causes of cancer death worldwide. This cancer exhibits high molecular and phenotype heterogeneity. The overall survival rate for gastric cancer is very low because it is always diagnosed in the advanced stages. Therefore, early detection and treatment are of great significance. Currently, biomedical studies have tapped the potential clinical applicability of aptamer-based technology for gastric cancer diagnosis and targeted therapy. Herein, we summarize the enrichment and evolution of relevant aptamers, followed by documentation of the recent developments in aptamer-based techniques for early diagnosis and precision therapy for gastric cancers.
Assuntos
Aptâmeros de Nucleotídeos , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/terapia , Medicina de Precisão , Aptâmeros de Nucleotídeos/uso terapêutico , Aptâmeros de Nucleotídeos/genética , TecnologiaRESUMO
Loading hyaluronidase (Hyal) in a nanocarrier is a potent strategy to degrade the tumor extracellular matrix for tumor deep penetration and enhanced tumor therapy. Herein, a pH-sensitive biomimicking nanosystem with high Hyal loading, effective tumor targeting, and controllable release is constructed. Specifically, cationic mesoporous silica nanoparticles (CMSNs) with large pores 13.52 nm in diameter were synthesized in a one-pot manner by adding N-[3-trimethoxysilylpropyl]-N,N,N-trimethylammonium to a reversed microemulsion reaction system. The Hyal loading rate was as high as 19.47% owing to matched pore size and the cationic surface charge. Subsequently, a pH-sensitive biomimetic hybrid membrane (pHH) composed of pH-sensitive liposome (pHL), red blood cell membrane, and pancreatic cancer cell membrane was camouflaged on the pHL-coated and doxorubicin/Hyal-loaded CMSNs (shortened as DHCM). The DHCM@pHL@pHH is stable at neutral pH while it releases the payloads smoothly in the tumor acidic microenvironment. Consequently, it can escape from macrophage clearance, be specifically taken up by pancreatic cancer cells, and efficiently accumulate at the tumor site. More importantly, it can penetrate deeply in pancreatic tumors with a tumor growth inhibition ratio of 80.46%. The nanosystem is biocompatible and has potential for clinical transformation, and the nanocarrier is promisingly applicable as a platform for encapsulation of various macromolecules for smart and tumor-targeted delivery.
Assuntos
Nanopartículas , Neoplasias Pancreáticas , Humanos , Dióxido de Silício/química , Hialuronoglucosaminidase , Sistemas de Liberação de Medicamentos , Biomimética , Nanopartículas/química , Doxorrubicina/química , Neoplasias Pancreáticas/tratamento farmacológico , Concentração de Íons de Hidrogênio , Portadores de Fármacos/química , Porosidade , Microambiente TumoralRESUMO
Cancer starvation/photothermal combined tumor therapy (CST/PTT) has attracted great interest attributed to their mutual compensation and synergistically enhanced effect. However, the very low O2 supply in the tumor microenvironment (TME) greatly limits the CST efficiency of glucose oxidase (GOx). Additionally, the easy degradation in blood circulation and significant off-target effects are big challenges for clinical applications of the GOx-based CST. In this study, a drug delivery system (DDS) with specific tumor-targeted GOx delivery, near-infrared (NIR) light and TME responsive O2 generation, NIR-responsive glucose consumption, high GOx loading, and efficient NIR photothermia was developed. Positively charged AuNRs@MnO2@SiO2 nanoparticles (named AMS+ NPs) were synthesized. GOx was covalently loaded with a high loading ratio of 36.0%. Finally, a thermosensitive biomimetic hybrid membrane composed of a thermosensitive lipid (TSL) membrane, red blood cell membrane (RBCM), and 4T1 cancer cell membrane (CCM) was coated on the NPs through a double-layer strategy. The AMS+-G@TSL@[RBC-CC-TSL]M NPs consumed 32.7 times glucose at 50 °C as that at 37 °C and generated 4.9 times O2 upon NIR laser irradiation. The thermosensitive biomimetic NPs showed an efficient targeting capability to the homotypic 4T1 cancer cells/tumors accompanied by good biocompatibility, macrophage evading capability, high cancer cell cytotoxicity, and excellent antitumor efficacy. The tumor growth inhibition ratio with NIR laser irradiation reached 92.8%. The AMS+-GOx@TSL@[RBC-CC-TSL]M NPs provide a smart, efficient, safe, PTT/CST combined DDS for highly efficient tumor therapy.
Assuntos
Biomimética , Neoplasias , Humanos , Compostos de Manganês , Óxidos , Dióxido de Silício , Glucose , Glucose Oxidase , Microambiente TumoralRESUMO
As an appealing biomimetic strategy for various medical applications, cell membrane coating lacks sensitive on-demand breaking capability. Herein, we incorporated thermosensitive lipid (TSL) membrane into red blood cell (RBC) and MCF-7 cancer cell (MC) hybrid membrane ([RBC-MC]M) vesicles. The [RBC-MC-TSL]M was coated onto doxorubicin (Dox)-loaded hollow gold nanoparticles to enhance chemo-/photothermal combined tumor therapy at a mild hyperthermia temperature (≤49 °C). Double-layer coating with TSL and [RBC-MC-TSL]M as the inner and outer layer, respectively, presented better antileakage and higher NIR-responsivity than single-layer coating. The Dox release ratio upon NIR laser irradiation (≤49 °C) was 74.6%, much higher than that (33.5%) without NIR laser. The nanodrug can be efficiently and specifically taken up by MCF-7 cells. In addition, the nanodrug exhibited excellent tumor-targeting property, with 4.08- and 1.12-times Dox accumulation in MCF-7 tumors compared to free Dox and [RBC-MC]M-coated counterpart, respectively. Most importantly, TSL incorporation significantly enhanced NIR-responsive antitumor efficiency, with tumor growth inhibition ratio increased from 35.1% to 48.6% after a single dose administration. Besides, the nanodrug exhibited very good biocompatibility. Camouflaging nanoparticles with the thermosensitive biomimetic hybrid membrane provides a painless and promisingly clinical-applicable approach for effective chemo-/photothermal combined mild-hyperthermia tumor therapy.
Assuntos
Hipertermia Induzida , Nanopartículas Metálicas , Ouro/farmacologia , Biomimética , Nanopartículas Metálicas/uso terapêutico , Doxorrubicina/farmacologiaRESUMO
Carcinoembryonic antigen (CEA) is an important malign tumor marker. In this study, a simple, label-free and antibody-free aptasensor was fabricated based on a multifunctional dendrimer-like DNA nanoassembly. The DNA nanoassembly was embedded with multiple G-quadruplex DNAzyme motifs and a hanging CEA aptamer motif. It was prepared from short DNA sequences by autonomous-assembly. The aptasensor was prepared simply by self-assembly of a capture DNA (cpDNA) on a gold electrode, followed by hybridization with a CEA aptamer (AptGAC-P). CEA as a model target was detected through competitive binding of CEA with AptGAC-P, exposing cpDNA to bind with the DNA nanoassembly. The detection process only contains 2 incubation steps. The high load of G-quadruplex DNAzyme motifs and their catalytic activity resulted in an amplified and label-free differential pulse voltammetry (DPV) electrochemical signal. The peak current correlated linearly with the CEA concentration, with a linear range of 2-45 ng mL-1, and an LOD value of 0.24 ng mL-1. The aptasensor showed high specificity and reproducibility, and retained 96.5% of detection signal intensities after 31 days of storage. The recovery rates for spiked CEA in human serum were within 100 ± 5%, and the coincidence rates for clinical human serum samples with ELISA kits were 80.7-111%. Conceivably, possessing simplicity, sensitivity, reproducibility, storage stability, and accuracy, the aptasensor should be a very prominent and applicable tool for clinical CEA detection and cancer diagnosis, and is promisingly applicable as a platform for detecting other targets of interests.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , DNA Catalítico , Dendrímeros , Antígeno Carcinoembrionário , Catálise , DNA , Humanos , Reprodutibilidade dos TestesRESUMO
The unique structures and properties of carbon nanotubes (CNTs) have attracted extensive investigations for many applications, such as those in the field of biomedical materials and devices, biosensors, drug delivery, and tissue engineering. Anticipated large-scale productions for numerous diversified applications of CNTs might adversely affect the environment and human health. For successful applications in the biomedical field, the issue of interfacing between CNTs and mammalian cells in vitro needs to be addressed before in vivo studies can be carried out systematically. We review the important studies pertaining to the internalization of CNTs into the cells and the culturing of cells on the CNT-based scaffold or support materials. The review will focus on the description of a variety of factors affecting CNT cytotoxicity: type of CNTs, impurities, lengths of CNTs, aspect ratios, dispersion, chemical modification, and assaying methods of cytotoxicity.
Assuntos
Nanotubos de Carbono/toxicidade , Técnicas Biossensoriais , Sistemas de Liberação de Medicamentos , Humanos , Nanotubos de Carbono/química , Engenharia TecidualRESUMO
In this study, carbon nanotubes (CNTs) were modified to further improve their performance in electrochemical sensing of dopamine (DA) levels. After a redox polymer, poly(vinylimidazole) complexed with Os(4, 4'-dimethyl- 2, 2-bipyridine)(2)Cl (termed PVI-dmeOs) was electrodeposited on multi-wall CNTs (MWCNTs), Nafion and PVI-dmeOs films were successfully layer-by-layer (LBL) assembled on the hydrophilic surface of the as-prepared PVI-dmeOs/CNTs nanocomposites through electrostatic interactions. The LBL assembly was proved by scanning electron microscopy (SEM), electrochemistry and UV-vis spectroscopy measurements. LBL assembly of Nafion/PVI-dmeOs films on CNTs significantly enhanced their linear sweep voltammetry (LSV) response sensitivity to DA, with a maximum enhancement for three Nafion/PVI-dmeOs film-modified MWCNTs. The LSV peak current density of (Nafion/PV I-dmeOs)(3)/CNT electrodes in response to 10 and 50 microM DA solutions was about 7.3 and 3.9 times those for bare CNTs. At the (Nafion/PV I-dmeOs)(3)/CNT electrodes, the limit of detection (LOD) (signal-to-noise ratio: 3) was 0.05 microM DA, the linear range was 0.1-10 microM DA (with a linear regression coefficient of 0.97) and the DA-sensing sensitivity was 8.15 microA cm( - 2) microM( - 1). The newly fabricated (Nafion/PV I-dmeOs)(3)/CNT electrodes may be developed as an ideal biosensor for direct and in situ measurement of DA levels.
Assuntos
Dopamina/análise , Polímeros de Fluorcarboneto/química , Nanotecnologia/métodos , Nanotubos de Carbono/química , 2,2'-Dipiridil/química , Técnicas Biossensoriais , Impedância Elétrica , Imidazóis/química , Modelos Lineares , Microscopia Eletrônica de Varredura , Compostos Organometálicos/química , Osmio/química , Polivinil/química , Sensibilidade e Especificidade , Espectrofotometria UltravioletaRESUMO
To clarify the endogenous pollution and release characteristics of the bottom sediment of Hengshan Reservoir in Yixing City, a typical section of the reservoir was sampled and analyzed. The research results show that the average concentrations of total nitrogen, total phosphorus, and organic matter in the surface sediments of Hengshan Reservoir are 2778 mg·kg-1, 899 mg·kg-1, and 3.1%, respectively. The endogenous pollution is serious, and the downstream sediments are highly polluted upstream of the reservoir. Phosphorus spectroscopic analysis results show that iron-bound phosphorus (Fe-P) and aluminum-bound phosphorus (Al-P) are the main bound phosphorus forms in the sediment, accounting for 28% and 39% of the total phosphorus, respectively. The average concentration of activated phosphorus in the sediment (combination of weakly adsorbed phosphorus, organic phosphorus, and iron phosphorus) is 255 mg·kg-1, accounting for 38% of the total phosphorus. The average release rates of nitrogen and phosphorus in sediments were 18.0 mg·(m2·d)-1 and 0.60 mg·(m2·d)-1. The correlation analysis results show that the organic matter content of the sediment is significantly correlated with the diffusion flux of phosphate, ammonia nitrogen, and ferrous iron (P<0.05), indicating that the mineralization of organic matter in the sediment may be the main release source of nitrogen and phosphorus in the sediment influencing factors.
RESUMO
DNA aptamers against carcinoembryonic antigen (CEA) have been identified through the systematic evolution of ligands by exponential enrichment (SELEX) technique, but their affinity needs to be improved. In this study, an in silico approach was firstly used to screen the mutation sequences of a reported DNA aptamer (the parent aptamer, denoted as P) against CEA. The affinities of several high-score DNA mutants were determined by the biolayer interferometry technique. Finally, the newly obtained aptamers were verified in an aptasensor application. For the in silico approach, Mfold and RNA Composer were combined to generate the 3D RNA structures of the DNA mutants. The RNA structures were then modified to 3D DNA structures with the Write program. The docking model and binding ability of the 3D DNA structures with CEA were simulated and predicted with the ZDOCK program. Two mutation sequences (P-ATG and GAC-P) exhibited significantly higher ZDOCK scores than P. The dissociation constant of P-ATG and GAC-P to CEA was determined to be 4.62 and 3.93 nM respectively, obviously superior to that of P (6.95 nM). The detection limit of the P-ATG and GAC-P based aptasensors was 1.5 and 1.2 ng mL-1, respectively, markedly better than that based on P (3.4 ng mL-1). The consistency between the in silico and the experimental results indicates that the developed in silico post-SELEX screening approach is feasible for improving DNA aptamers. The P-ATG and GAC-P aptamers found in this study could be used for future CEA aptasensor design and fabrication, promisingly applicable for highly sensitive CEA detection and early cancer diagnosis.
RESUMO
A stable and sensitive electrochemical acetylcholinesterase (AChE) biosensor for detection of organophosphorus pesticides (OPs) was developed by doping Au nanorods (AuNRs)@mesoporous SiO2 (MS) core-shell nanoparticles into CS/TiO2-CS (CS denotes for chitosan) immobilization matrix. AuNRs@MS core-shell nanoparticles were synthesized and characterized. The doping and the biosensor fabrication process were probed and confirmed by scanning electron microscopy and electrochemistry techniques. The doping conditions were optimized. The matrix both before and after AChE immobilization had a mesoporous nanostructure. The nanoparticles dispersed homogeneously within the matrix. The doping significantly enhanced the electro-conductivity of the TiO2-CS hydrogel, and dramatically improved the bioelectrocatalytic activity and OPs detection sensitivity of the AChE immobilized matrix. The detection linear ranges for both dichlovos (DDVP) and fenthion were from 0.018⯵M (4.0â¯ppb) to 13.6⯵M, and the limit of detection (LOD) was 5.3â¯nM (1.2â¯ppb) and 1.3â¯nM (0.36â¯ppb), respectively. The biosensor exhibited high reproducibility and accuracy in detecting OPs spiked vegetable juice samples. In addition, it exhibited very high detection stability and storage stability. The developed AChE biosensor was provided to be a promisingly applicable tool for OPs detection with high reliability, simplicity, and rapidness.