RESUMO
BACKGROUND Emergence cough during endotracheal extubation can lead to complications. This cough is triggered by the deviation of the endotracheal tube from the airway anatomy, causing pressure on the airway mucosal wall. Head elevation has the potential to align the airway passage with the tube's configuration. In this study, we aimed to investigate the impact of head elevation using a pillow on the prevalence and severity of emergence cough in male patients. MATERIAL AND METHODS A total of 71 male patients undergoing laparoscopic cholecystectomy were randomly assigned to either the head elevation group (n=35) or the control group (n=36). The head elevation group maintained a position with a neck flexion angle of 35º using a pillow, while the control group remained in a neutral position after anesthetic induction. The severity of cough was assessed before, during, and after extubation using a 4-point scale, with grades 2 and 3 indicating cough and grade 3 indicating severe cough. RESULTS The characteristics and intraoperative data of the patients were similar between the two groups. There was no significant difference in the incidence of cough and severe cough between the groups. However, the severity of cough was significantly lower in the head elevation group compared to the control group before extubation (cough scale: 0/5/8/23 vs 1/2/17/15 in the control group vs the head elevation group, P=0.039). The time to extubation, respiratory complications, nausea, pain, and the number of patients receiving fentanyl were comparable between the groups. CONCLUSIONS Head elevation using a pillow effectively reduced the severity of cough before endotracheal extubation during anesthesia emergence in male patients. However, it did not significantly reduce the incidence of cough. These findings highlight the potential benefits of head elevation in minimizing the discomfort associated with emergence cough.
Assuntos
Extubação , Tosse , Humanos , Masculino , Extubação/efeitos adversos , Tosse/etiologia , Tosse/epidemiologia , Prevalência , Fentanila , Dor/complicações , Intubação Intratraqueal/efeitos adversosRESUMO
The MS2 system, with an MS2 binding site (MBS) and an MS2 coat protein fused to a fluorescent protein (MCP-FP), has been widely used to fluorescently label mRNA in live cells. However, one of its limitations is the constant background fluorescence signal generated from free MCP-FPs. To overcome this obstacle, we used a superfolder GFP (sfGFP) split into two or three nonfluorescent fragments that reassemble and emit fluorescence only when bound to the target mRNA. Using the high-affinity interactions of bacteriophage coat proteins with their corresponding RNA binding motifs, we showed that the nonfluorescent sfGFP fragments were successfully brought close to each other to reconstitute a complete sfGFP. Furthermore, real-time mRNA dynamics inside the nucleus as well as the cytoplasm were observed by using the split sfGFPs with the MS2-PP7 hybrid system. Our results demonstrate that the split sfGFP systems are useful tools for background-free imaging of mRNA with high spatiotemporal resolution.
Assuntos
Proteínas de Fluorescência Verde/ultraestrutura , Imagem Molecular/métodos , RNA Mensageiro/ultraestrutura , Linhagem Celular , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Proteínas de Fluorescência Verde/genética , Humanos , Hibridização in Situ Fluorescente , RNA Mensageiro/genéticaRESUMO
In situ wireless monitoring for cell proliferation and detachment kinetics was conducted using pH-responsive zwitterionic polymer dots (Z-PDs), based on changes in electrochemical signals derived from Z-PD-coated substrates via the interaction of charges transferred between Z-PDs and cells. Z-PD-coated substrates were found to be a potent means to monitor and manipulate cell adhesion and detachment because of their high sensitivity over a wide range of pH conditions, and modification of the coated substrates was confirmed using a wireless system. At neutral pH, Z-PD-coated wireless sensors exhibited π-π stacking involving aromatic rings with hydrophobic interactions, thereby promoting cell proliferation; consequently, an increase in the measured resistance was observed. In contrast, Z-PD-coated substrates triggered by acidic and basic conditions promoted cell detachment, which induced an increase in the resistance compared with Z-PD substrates at pH 6.8, as a result of charges transferred to support Z-PD internalization through cell membranes after detachment. Therefore, as a wireless biosensor with excellent pH responsiveness that facilitates cell proliferation and detachment and whose electrochemical signals could be additionally acquired via a smartphone, Z-PD biosensors demonstrated a more favorable approach for monitoring cell-surface interactions than conventional optically based methods.
Assuntos
Polímeros , Proliferação de Células , Condutividade Elétrica , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e HidrofílicasRESUMO
Previous work has demonstrated that fusion of a luciferase to an opsin, to create a luminescent opsin or luminopsin, provides a genetically encoded means of manipulating neuronal activity via both chemogenetic and optogenetic approaches. Here we have expanded and refined the versatility of luminopsin tools by fusing an alternative luciferase variant with high light emission, Gaussia luciferase mutant GLucM23, to depolarizing and hyperpolarizing channelrhodopsins with increased light sensitivity. The combination of GLucM23 with Volvox channelrhodopsin-1 produced LMO4, while combining GLucM23 with the anion channelrhodopsin iChloC yielded iLMO4. We found efficient activation of these channelrhodopsins in the presence of the luciferase substrate, as indicated by responses measured in both single neurons and in neuronal populations of mice and rats, as well as by changes in male rat behavior during amphetamine-induced rotations. We conclude that these new luminopsins will be useful for bimodal opto- and chemogenetic analyses of brain function.
Assuntos
Channelrhodopsins , Luciferases , Neurônios/fisiologia , Optogenética/métodos , Potenciais de Ação , Adenoviridae/fisiologia , Animais , Channelrhodopsins/genética , Channelrhodopsins/fisiologia , Feminino , Vetores Genéticos , Células HEK293 , Hipocampo/fisiologia , Humanos , Luciferases/genética , Luciferases/fisiologia , Masculino , Camundongos , Cultura Primária de Células , Ratos Sprague-Dawley , Volvox/genéticaRESUMO
Luminopsins are fusion proteins of luciferase and opsin that allow interrogation of neuronal circuits at different temporal and spatial resolutions by choosing either extrinsic physical or intrinsic biological light for its activation. Building on previous development of fusions of wild-type Gaussia luciferase with channelrhodopsin, here we expanded the utility of luminopsins by fusing bright Gaussia luciferase variants with either channelrhodopsin to excite neurons (luminescent opsin, LMO) or a proton pump to inhibit neurons (inhibitory LMO, iLMO). These improved LMOs could reliably activate or silence neurons in vitro and in vivo. Expression of the improved LMO in hippocampal circuits not only enabled mapping of synaptic activation of CA1 neurons with fine spatiotemporal resolution but also could drive rhythmic circuit excitation over a large spatiotemporal scale. Furthermore, virus-mediated expression of either LMO or iLMO in the substantia nigra in vivo produced not only the expected bidirectional control of single unit activity but also opposing effects on circling behavior in response to systemic injection of a luciferase substrate. Thus, although preserving the ability to be activated by external light sources, LMOs expand the use of optogenetics by making the same opsins accessible to noninvasive, chemogenetic control, thereby allowing the same probe to manipulate neuronal activity over a range of spatial and temporal scales.
Assuntos
Luz , Opsinas/metabolismo , Optogenética , Potenciais de Ação/efeitos da radiação , Animais , Comportamento Animal , Feminino , Células HEK293 , Humanos , Luciferases/metabolismo , Medições Luminescentes , Camundongos Endogâmicos C57BL , Movimento , Neurônios/metabolismo , Neurônios/efeitos da radiação , Ratos Sprague-Dawley , Rodopsina/metabolismo , Substância Negra/fisiologia , Substância Negra/efeitos da radiação , Sinapses/metabolismo , Sinapses/efeitos da radiação , Volvox/metabolismo , Volvox/efeitos da radiaçãoRESUMO
Herein, we describe the fabrication and characterization of carbonized disulfide core-crosslinked polymer dots with pH-cleavable colorimetric nanosensors, based on diol dye-conjugated fluorescent polymer dots (L-PD), for reduction-triggered paclitaxel (PTX) release during fluorescence imaging-guided chemotherapy of tumors. L-PD were loaded with PTX (PTX loaded L-PD), via π-π stackings or hydrophobic interactions, for selective theragnosis by enhanced release of PTX after the cleavage of disulfide bonds by high concentration of glutathione (GSH) in a tumor. The nano-hybrid system showed fluorescence quenching behavior with less than 2% of PTX released under physiological conditions. However, in a tumor microenvironment, the fluorescence recovered at an acidic-pH, and PTX (approximately 100% of the drug release) was released efficiently out of the matrix by reduction caused by the GSH level in the tumor cells, which improved the effectiveness of the cancer treatment. Therefore, the colorimetric nanosensor showed promising potential in distinguishing between normal and cancerous tissues depending on the surrounding pH and GSH concentrations so that PTX can be selectively delivered into cancer cells for improved cancer diagnosis and chemotherapy.
Assuntos
Antineoplásicos/administração & dosagem , Nanopartículas/química , Paclitaxel/administração & dosagem , Polímeros/química , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Técnicas Biossensoriais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Colorimetria , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Fluorescência , Glutationa/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Nanopartículas/ultraestrutura , Oxirredução , Paclitaxel/síntese química , Paclitaxel/química , Paclitaxel/uso terapêuticoRESUMO
A novel mechanochemical method was firstly developed to synthesize carbon nanodots (CNDs) or carbon nano-onions (CNOs) through high-pressure homogenization of cellulose powders as naturally abundant resource depending on the treatment times. While CNDs (less than 5 nm in size) showed spherical and amorphous morphology, CNOs (10-50 nm in size) presented polyhedral shape, and onion-like outer lattice structure, graphene-like interlattice spacing of 0.36 nm. CNOs showed blue emissions, moderate dispersibility in aqueous media, and high cell viability, which enables efficient fluorescence imaging of cellular media.
RESUMO
This work derived biocompatible and stable probes based on fluorescent nanoparticles (FNPs) from a natural source, Curcuma longa. The multi-color fluorescence emissions from carbonized Curcuma longa (C-FNPs) obtained through defined dehydration conditions are soluble in water and have a small particle size (~17 nm). The surface passivation with polyethylene glycol (PEG) capped with amine groups in FNPs (P-FNPs) generated a probe with a higher quantum yield and longer fluorescence lifetime than obtained with C-FNPs. The X-ray photoelectron spectroscopy and X-ray diffraction spectra confirmed the associated chemical moieties of C-FNPs and P-FNPs. Furthermore, the prepared material showed non-toxic effects with almost 100% cell viability, even at high concentrations. In conclusion, fluorescence sensors from natural sources may be useful for numerous biomedical research applications.
Assuntos
Materiais Biocompatíveis/química , Carbono/química , Curcuma/química , Corantes Fluorescentes/química , Nanopartículas/química , Pontos Quânticos , Animais , Sobrevivência Celular , Cães , Humanos , Células KB , Células Madin Darby de Rim Canino , Tamanho da Partícula , Espectrometria de Fluorescência , Propriedades de SuperfícieRESUMO
The tumor-specific sensitive fluorescence sensing of cellular alkaline phosphatase (ALP) activity on the basis of host-guest specific and pH sensitivity was conducted on coated surfaces and aqueous states. Cross-linked fluorescent nanoparticles (C-FNP) consisting of ß-cyclodextrin (ß-CD)/boronic acid (BA) and fluorescent hyaluronic acid [FNP(HA)] were conjugated to fluorescent polydopamine [FNP(pDA)]. To determine the quenching effect of this system, hydrolysis of 4-nitrophenyl phosphate (NPP) to 4-nitrophenol (NP) was performed in the cavity of ß-CD in the presence of ALP activated photoinduced electron transfer (PET) between NP and C-FNP. At an ALP level of 30-1000 U/L, NP caused off-emission of C-FNP because of their specific host-guest recognition. Fluorescence can be recovered under pH shock due to cleavage of the diol bond between ß-CD and BA, resulting in release of NP from the fluorescent system. Sensitivity of the assays was assessed by confocal imaging not only in aqueous states, but also for the first time on coated surfaces in MDAMB-231 and MDCK cells. This novel system demonstrated high sensitivity to ALP through generation of good electron donor/acceptor pair during the PET process. Therefore, this fluorescence sensor system can be used to enhance ALP monitoring and cancer diagnosis on both coated surfaces and in aqueous states in clinical settings.
Assuntos
Fosfatase Alcalina/metabolismo , Carbono/química , Reagentes de Ligações Cruzadas/química , Corantes Fluorescentes/química , Indóis/química , Nanopartículas/química , Polímeros/química , Animais , Linhagem Celular Tumoral , Cães , Fluorescência , Humanos , Concentração de Íons de Hidrogênio , Células Madin Darby de Rim Canino , Nitrofenóis/química , Nitrofenóis/metabolismo , Compostos Organofosforados/química , Compostos Organofosforados/metabolismo , Tamanho da Partícula , Soluções , Propriedades de Superfície , Água/química , beta-Ciclodextrinas/química , beta-Ciclodextrinas/metabolismoRESUMO
In cancer therapy, optimizing tumor-specific delivery, tumor distribution, and cellular uptake of a drug is important for ensuring minimal toxicity and maximum therapeutic efficacy. This study characterized the therapeutic efficacy of a stimulus responsive and dual targeting nanocarrier for a bioimaging-guided photothermal and chemotherapeutic platform. Hyaluronic acid (HA) conjugated with triphenylphosphonium (TPP) and boronic acid (BA) diol-linked ß-cyclodextrin (ß-CD) forms an inclusion complex with paclitaxel (PTX), creating a shell-like composite on a core of carbonized fluorescent polydopamine nanoparticles (FNPs-pDA) applicable for photothermal therapy as well as bioimaging. The successful diol cross-linking between core@shells generates nanocarriers [FNPs-pDA@HA-TPP-CD-PTX] that can be used as an extracellular HA- and intracellular TPP-mediated dual targeting system. The carbonized FNPs-pDA was cross-linked with the boronic acid groups of HA-TPP-CD-PTX to promote the formation of boronate esters for pH-mediated photothermal activity, which have shown time dependent complete PTX release along with a photothermal mediated response. The in vitro dual bioimaging and photothermal-chemotherapeutic activities were compared between cancer and normal cells. Lysosomal escape and live/dead cells staining confocal images highlight the promise of this system, which might open up a new approach, a simple and versatile method for site-specific synergetic drug delivery.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Portadores de Fármacos , Ácido Hialurônico/química , Indóis/química , Nanopartículas/química , Paclitaxel/farmacologia , Polímeros/química , Animais , Ácidos Borônicos/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cães , Composição de Medicamentos/métodos , Humanos , Receptores de Hialuronatos/metabolismo , Concentração de Íons de Hidrogênio , Raios Infravermelhos , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Células Madin Darby de Rim Canino , Nanopartículas/metabolismo , Nanopartículas/ultraestrutura , Oniocompostos/química , Processos Fotoquímicos , Compostos de Tritil/química , beta-Ciclodextrinas/químicaRESUMO
Polymer dots (PDs) showing concentration-mediated multicolor fluorescence were first prepared from sulfuric acid-treated dehydration of Pluronic® F-127 in a single step. Pluronic-based PDs (P-PDs) showed high dispersion stability in solvent media and exhibited a fluorescence emission that was widely tunable from red to blue by adjusting both the excitation wavelengths and the P-PD concentration in an aqueous solution. This unique fluorescence behavior of P-PDs might be a result of cross-talk in the fluorophores of the poly(propylene glycol)-rich core inside the P-PD through either energy transfer or charge transfer. Reconstruction of the surface energy traps of the P-PDs mediated through aggregation may lead to a new generation of carbon-based nanomaterials possessing a fluorescence emission and tunable by adjusting the concentration. These structures may be useful in the design of multifunctional carbon nanomaterials with tunable emission properties according to a variety of internal or external stimuli.
Assuntos
Carbono/química , Fluorescência , Corantes Fluorescentes/química , Poloxâmero/química , Polímeros/química , Propilenoglicóis/química , Corantes Fluorescentes/síntese química , Poloxâmero/síntese química , Polímeros/síntese química , Propilenoglicóis/síntese químicaRESUMO
Despite the tremendous progress that photothermal therapy (PTT) has recently achieved, it still has a long way to go to gain the effective targeted photothermal ablation of tumor cells. Driven by this need, we describe a new class of targeted photothermal therapeutic agents for cancer cells with pH responsive bioimaging using near-infrared dye (NIR) IR825, conjugated poly(ethylene glycol)-g-poly(dimethylaminoethyl methacrylate) (PEG-g-PDMA, PgP), and hyaluronic acid (HA) anchored reduced graphene oxide (rGO) hybrid nanoparticles. The obtained rGO nanoparticles (PgP/HA-rGO) showed pH-dependent fluorescence emission and excellent near-infrared (NIR) irradiation of cancer cells targeted in vitro to provide cytotoxicity. Using intravenously administered PTT agents, the time-dependent in vivo tumor target accumulation was exactly defined, presenting eminent photothermal conversion at 4 and 8 h post-injection, which was demonstrated from the ex vivo biodistribution of tumors. These tumor environment responsive hybrid nanoparticles generated photothermal heat, which caused dominant suppression of tumor growth. The histopathological studies obtained by H&E staining demonstrated complete healing from malignant tumor. In an area of limited successes in cancer therapy, our translation will pave the road to design stimulus environment responsive targeted PTT agents for the safe eradication of devastating cancer.
Assuntos
Grafite/química , Nanopartículas/química , Neoplasias/terapia , Fototerapia/métodos , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Linhagem Celular Tumoral , Cães , Grafite/farmacocinética , Humanos , Ácido Hialurônico/química , Ácido Hialurônico/farmacocinética , Concentração de Íons de Hidrogênio , Células Madin Darby de Rim Canino , Metacrilatos/química , Metacrilatos/farmacocinética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Nanocompostos/química , Polietilenoglicóis/química , Polietilenoglicóis/farmacocinética , Espectroscopia de Luz Próxima ao Infravermelho , Distribuição TecidualRESUMO
Gold nanoparticle (AuNP)-decorated boron nitride nanosheet (BNNS) was successfully prepared through the simultaneous reduction of Au(3+) ions and the growth of AuNPs on polydopamine (PDA)-grafted BNNS. Both BNNS-AuNP and PDA-BNNS are successfully synthesized in an aqueous buffer solution (pH 8.5) in the absence of any chemical reducing agent and organic reaction, which is therefore environmentally friendly and highly beneficial for the mass production of green catalysts from 2D nanomaterials. BNNS-AuNP showed remarkable dispersion stability in aqueous media and revealed high catalytic efficiency for the reduction of nitrophenol as (4-NP) into 4-aminophenol (4-AP) within 8 min in water. The 2D structural feature of BNNS-AuNP also enables isolation and recycling of catalyst from 4-AP through the ultracentrifugation, which shows the retention of more than 60% of catalytic activity of BNNS-AuNP after five repetitions of the of recycling steps.
RESUMO
Boron nitride nanosheet (BNNS) decorated with silver nanoparticles (AgNPs) was successfully synthesized via mussel-inspired chemistry of dopamine. Poly(dopamine)-functionalized BNNS (PDA-BNNS) was prepared by adding dopamine into the aqueous dispersion of hydroxylated BNNS (OH-BNNS) at alkaline condition. AgNPs were decorated on PDA-BNNS through spontaneous reduction of silver cations by catechol moieties of a PDA layer on BNNS, resulting in AgNP-BNNS with good dispersion stability. Incorporation of PDA on BNNS not only played a role as a surface functionalization method of BNNS, but also provided a molecular platform for creating very sophisticated two-dimensional (2D) BNNS-based hybrid nanomaterials such as metal nanoparticle-decorated BNNS.
Assuntos
Compostos de Boro/química , Dopamina/química , Nanopartículas/química , Prata/química , Antibacterianos/química , Antibacterianos/uso terapêutico , Humanos , Hidroxilação , Microscopia de Força Atômica , Microscopia Eletrônica de Transmissão , Nanopartículas/uso terapêutico , Nanopartículas/ultraestruturaRESUMO
A coenzyme A (CoA-SH)-responsive dual electrochemical and fluorescence-based sensor was designed utilizing an MnO2-immobilized-polymer-dot (MnO2@D-PD)-coated electrode for the sensitive detection of osteoarthritis (OA) in a peroxisomal ß-oxidation knockout model. The CoA-SH-responsive MnO2@D-PD-coated electrode interacted sensitively with CoA-SH in OA chondrocytes, triggering electroconductivity and fluorescence changes due to cleavage of the MnO2 nanosheet on the electrode. The MnO2@D-PD-coated electrode can detect CoA-SH in immature articular chondrocyte primary cells, as indicated by the significant increase in resistance in the control medium (R24h = 2.17 MΩ). This sensor also sensitively monitored the increase in resistance in chondrocyte cells in the presence of acetyl-CoA inducers, such as phytol (Phy) and sodium acetate (SA), in the medium (R24h = 2.67, 3.08 MΩ, respectively), compared to that in the control medium, demonstrating the detection efficiency of the sensor towards the increase in the CoA-SH concentration. Furthermore, fluorescence recovery was observed owing to MnO2 cleavage, particularly in the Phy- and SA-supplemented media. The transcription levels of OA-related anabolic (Acan) and catabolic factors (Adamts5) in chondrocytes also confirmed the interaction between CoA-SH and the MnO2@D-PD-coated electrode. Additionally, electrode integration with a wireless sensing system provides inline monitoring via a smartphone, which can potentially be used for rapid and sensitive OA diagnosis.
Assuntos
Técnicas Biossensoriais , Eletrodos , Compostos de Manganês , Osteoartrite , Polímeros , Compostos de Manganês/química , Polímeros/química , Óxidos/química , Técnicas Eletroquímicas , Oxirredução , Condrócitos , Humanos , Fluorescência , Peroxissomos/metabolismo , AnimaisRESUMO
An electrochemical sensor sensitive to coenzyme A (CoA) was designed using a CoA-responsive polyallylamine-manganese oxide-polymer dot nanogel coated on the electrode surface to detect various genetic models of osteoarthritis (OA). The CoA-responsive nanogel sensor responded to the abundance of CoA in OA, causing the breakage of MnO2 in the nanogel, thereby changing the electroconductivity and fluorescence of the sensor. The CoA-responsive nanogel sensor was capable of detecting CoA depending on the treatment time and distinguishing the response towards different OA genetic models that contained different levels of CoA (wild type/WT, NudT7 knockout/N7KO, and Acot12 knockout/A12KO). The WT, N7KO, and A12KO had distinct resistances, which further increased as the incubation time were changed from 12 h (R12h = 2.11, 2.40, and 2.68 MΩ, respectively) to 24 h (R24h = 2.27, 2.59, and 2.92 MΩ, respectively) compared to the sensor without treatment (Rcontrol = 1.63 MΩ). To simplify its application, the nanogel sensor was combined with a wireless monitoring device to allow the sensing data to be directly transmitted to a smartphone. Furthermore, OA-indicated anabolic (Acan) and catabolic (Adamts5) factor transcription levels in chondrocytes provided evidence regarding CoA and nanogel interactions. Thus, this sensor offers potential usage in simple and sensitive OA diagnostics.
RESUMO
Rapid and accurate diagnosis is crucial for managing the global health threat posed by multidrug-resistant bacterial infections; however, current methods have limitations in either being time-consuming, labor-intensive, or requiring instruments with high costs. Addressing these challenges, we introduce a wireless electrochemical sensor integrating the CRISPR/Cas system with electroconductive polymer dot (PD) nanoparticles to rapidly detect bacterial pathogens from human sputum. To enhance the electroconductive properties, we synthesized copper-ion-immobilized PD (PD-Cu), followed by conjugation of the deactivated Cas9 protein (dCas9) onto PD-Cu-coated Si electrodes to generate the dCas9-PD-Cu sensor. The dCas9-PD-Cu sensor integrated with isothermal amplification can specifically detect target nucleic acids of multidrug-resistant bacteria, such as the antibiotic resistance genes kpc-2 and mecA. The dCas9-PD-Cu sensor exhibits high sensitivity, allowing for the detection of â¼54 femtograms of target nucleic acids, based on measuring the changes in resistivity of the Si electrodes through target capture by dCas9. Furthermore, a wireless sensing platform of the dCas9-PD-Cu sensor was established using a Bluetooth module and a microcontroller unit for detection using a smartphone. We demonstrate the feasibility of the platform in diagnosing multidrug-resistant bacterial pneumonia in patients' sputum samples, achieving 92% accuracy. The current study presents a versatile biosensor platform that can overcome the limitations of conventional diagnostics in the clinic.
Assuntos
Ácidos Nucleicos , Pneumonia Bacteriana , Humanos , Polímeros , Cobre/química , Resistência a Múltiplos MedicamentosRESUMO
This work demonstrates the way to achieve efficient and target specific delivery of a graphene quantum dot (GQD) using hyaluronic acid (HA) (GQD-HA) as a targeting agent. HA has been anchored to a GQD that accepts the fascinating adhesive properties of the catechol moiety, dopamine hydrochloride, conjugated to HA, which was confirmed by X-ray photoelectron spectroscopy. Transmission electron microscopy revealed a particle size of â¼20 nm, and the fluorescence spectra revealed significant fluorescence intensity even after the anchoring of HA. The prepared GQD-HA was applied to CD44 receptor overexpressed tumor-bearing balb/c female mice, and the in vivo biodistribution investigation demonstrated more bright fluorescence from the tumor tissue. In vitro cellular imaging, via a confocal laser scanning microscope, exhibited strong fluorescence from CD44 overexpressed A549 cells. Both in vivo and in vitro results showed the effectiveness of using HA as targeting molecule. The loading and release kinetics of the hydrophobic drug doxorubicin from a GQD under mildly acidic conditions showed that a GQD can be considered as a novel drug carrier, while the nontoxic behavior from the MTT assay strongly supports the identification of GQD-HA as a biocompatible material.
Assuntos
Diagnóstico por Imagem/métodos , Grafite/química , Ácido Hialurônico/química , Pontos Quânticos/química , Animais , Materiais Biocompatíveis/química , Linhagem Celular , Cães , Dopamina/química , Portadores de Fármacos/química , Humanos , Receptores de Hialuronatos/metabolismo , Camundongos , Camundongos Endogâmicos BALB CRESUMO
This present article demonstrates the strategy to prepare photoresponsive reduced graphene oxide with mussel inspired adhesive material dopamine (DN) and photochromic dye spiropyran (SP) conjugated to the backbone of the targeting ligand hyaluronic acid (HA; HA-SP). Graphene oxide (GO) was reduced by prepared HA-SP accepting the advantages of catechol chemistry under mildly alkaline condition enabling to achieve functionalized graphene (rGO/HA-SP) as fluorescent nanoparticles. Due to containing HA, rGO/HA-SP can bind to the CD44 cell receptors. The prepared rGO/HA-SP is able to retain its photochromic features and can be converted to merocyanine (MC) form upon irradiation with UV light (wavelength: 365 nm) displaying purple color. Photochromic behavior of rGO/HA-SP was monitored by UV-vis and fluorescence spectroscopy. In vitro fluorescence behavior, examined by confocal laser scanning microscope (CLSM), of rGO/HA-SP in cancerous A549 cell lines assured that efficient delivery of rGO/HA-SP was gained due to HA as targeting ligand. In this work, we have shown that in vivo fluorescence image of spiropyran is possible by administrating MC form solution of rGO/HA-SP using Balb/C mice as in vivo modal. Accumulation of rGO/HA-SP in tumor tissue from biodistribution analysis strongly supports the specific delivery of prepared graphene to the target destination. The well tuned drug release manner from the surface of rGO/HA-SP strongly recommends the developed material not only as fluorescent probe for diagnosis but also as a drug carrier in drug delivery system.
Assuntos
Benzopiranos/química , Sistemas de Liberação de Medicamentos , Fluorescência , Grafite/química , Ácido Hialurônico/química , Indóis/química , Imagem Molecular , Nitrocompostos/química , Óxidos/química , Animais , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/farmacocinética , Benzopiranos/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Dopamina/química , Doxorrubicina/administração & dosagem , Doxorrubicina/farmacocinética , Humanos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/toxicidade , Indóis/toxicidade , Substâncias Luminescentes/química , Camundongos , Camundongos Endogâmicos BALB C , Nanopartículas/química , Nitrocompostos/toxicidade , Oxirredução , Processos Fotoquímicos , Distribuição Tecidual , Raios UltravioletaRESUMO
Herein, a cancer-specific dopamine-conjugated sp2-rich carbonized polymer dot (PD)-encapsulated mesoporous MnO2 (MnO2@PD)-mineralized hydrogel biosensor was developed that offers cancer-induced observable in situ alterations in fluorescence (FL), electrochemical, and mechanophysical properties. Cancer-triggered MnO2 degradation in the hydrogel, prompted by increased levels of glutathione (GSH) and reactive oxygen species (ROS) such as H2O2, leads to PD release and FL restoration, thereby controlling changes in the pore structure and increasing hydrogen bonding, resulting in physiologically visible alterations in mechanical stretchability, viscosity, swelling behavior, and adhesiveness. The pore size of the matrix increased from 21.83 to 36.81 m2/g upon GSH treatment, affecting the viscosity and swellability of the system. The resistance increased from 21.96 ± 1.16 to 30.69 ± 2.01 and 32.21 ± 2.54 kΩ, respectively, confirming the dependence of conductivity changes on H2O2 and GSH treatments. The in vitro treatment with cancer cells (HeLa, PC-3, and B16F10) facilitated a tunable electrochemical sensing performance via redox-mediated MnO2 breakdown by intracellular ROS and GSH, whereas hydrogels treated with normal cells (CHO-K1) showed minimal changes. Cancer-microenvironment-derived water-drop sensing showed three times higher response as compared to the normal cell-treated hydrogel. The sensing capability of the fabricated sensor was validated based on bending-induced relative resistance changes under dry and wet conditions. Moreover, the integration of the developed sensor with a wireless sensor enabled real-time monitoring with a smartphone.