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In this study, we developed a microfluidic device that is able to monitor cell biology under continuous PM2.5 treatment. The effects of PM2.5 on human alveolar basal epithelial cells, A549 cells, and uncovered several significant findings were investigated. The results showed that PM2.5 exposure did not lead to a notable decrease in cell viability, indicating that PM2.5 did not cause cellular injury or death. However, the study found that PM2.5 exposure increased the invasion and migration abilities of A549 cells, suggesting that PM2.5 might promote cell invasiveness. Results of RNA sequencing revealed 423 genes that displayed significant differential expression in response to PM2.5 exposure, with a particular focus on pathways associated with the generation of reactive oxygen species (ROS) and mitochondrial dysfunction. Real-time detection demonstrated an increase in ROS production in A549 cells after exposure to PM2.5. JC1 assay, which indicated a loss of mitochondrial membrane potential (ΔΨm) in A549 cells exposed to PM2.5. The disruption of mitochondrial membrane potential further supports the detrimental effects of PM2.5 on A549 cells. These findings highlight several adverse effects of PM2.5 on A549 cells, including enhanced invasion and migration capabilities, altered gene expression related to ROS pathways, increased ROS production and disruption of mitochondrial membrane potential. These findings contribute to our understanding of the potential mechanisms through which PM2.5 can impact cellular function and health.
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Movimiento Celular , Supervivencia Celular , Neoplasias Pulmonares , Potencial de la Membrana Mitocondrial , Material Particulado , Especies Reactivas de Oxígeno , Humanos , Material Particulado/efectos adversos , Especies Reactivas de Oxígeno/metabolismo , Células A549 , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/genética , Movimiento Celular/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Dispositivos Laboratorio en un Chip , Mitocondrias/metabolismo , Mitocondrias/efectos de los fármacos , Invasividad Neoplásica/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Microfluídica/métodosRESUMEN
Nanomaterials, specifically metal nanoclusters (NCs), are gaining attention as a promising class of antibacterial agents. Metal NCs exhibit antibacterial properties due to their ultrasmall size, extensive surface area, and well-controlled surface ligands. The antibacterial mechanisms of metal NCs are influenced by two primary factors: size and surface charge. In this review, we summarize the impacts of size and surface charge of metal NCs on the antibacterial mechanisms, their interactions with bacteria, and the factors that influence their antibacterial effects against both gram-negative and gram-positive bacteria. Additionally, we highlight the mechanisms that occur when NCs are negatively or positively charged, and provide examples of their applications as antibacterial agents. A better understanding of relationships between antibacterial activity and the properties of metal NCs will aid in the design and synthesis of nanomaterials for the development of effective antibacterial agents against bacterial infections. Based on the remarkable achievements in the design of metal NCs, this review also presents conclusions on current challenges and future perspectives of metal NCs for both fundamental investigations and practical antibacterial applications.
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Nanopartículas del Metal , Antibacterianos/farmacología , OroRESUMEN
Surface plasmon resonance (SPR) nanowire array chips with a microfluidic system are an effective detection method for a rapid test device. This study investigated a capped gold nanowire array and a microfluidic test platform to provide a fundamental understanding of the kinetic binding of SPR nanowires and the surface gold refractive index. The device sensitivity of the SPR nanowire array was 485 nm RIU-1 and the detection limit was 4.1 × 10-5 RIU. Moreover, a kinetic binding analysis also indicated that a peak shift resulted from a specific hybridization of the target molecule with the immobilized probe on the gold nanostructures. The peak shift (red-shift) value of latent membrane protein 1 (LMP1) DNA was 2.21 nm. The results demonstrated that this new method had high sensitivity to detect amplified DNA products without labeling or complex sample treatment. The SPR nanowire chip can detect the PCR products at lower cycle numbers compared to gel electrophoresis due to probe and DNA specificity. Furthermore, the mechanisms of SPR nanowire array fabrication and the detection of the LMP1 gene were studied. The findings can assist in improving the biosensing of DNA-amplified products and in developing rapid detection devices with a small-footprint nanostructured SPR chip.
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ADN Viral/análisis , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas/métodos , Nanocables/química , Resonancia por Plasmón de Superficie/métodos , Proteínas de la Matriz Viral/genética , Secuencia de Bases , Técnicas Biosensibles/métodos , Sondas de ADN/química , Sondas de ADN/genética , ADN Viral/genética , Oro/química , Herpesvirus Humano 4/química , Ácidos Nucleicos Inmovilizados/química , Ácidos Nucleicos Inmovilizados/genética , Límite de Detección , Técnicas Analíticas Microfluídicas/instrumentación , Hibridación de Ácido NucleicoRESUMEN
In recent years, the development of biodegradable hydrogels as an alternative over the traditional wound dressing has become increasingly significant. These specific hydrogels are able to offer suitable microenvironments to further aid the process of tissue or organ regeneration. However, application of biodegradable hydrogels in clinical medicine remains uncommon due to most biodegradable hydrogels struggle with achieving satisfactory adhesiveness property, high mechanical support and cell compatibility simultaneously. In order to overcome these constraints and enhance the applicability of biodegradable hydrogels, methods have been employed in this study. By reacting gellan gum with methacrylic anhydride and incorporating a biodegradable protein, keratin, we endowed the hydrogels with high pliability via photo-polymerization chain extension, thereby obtaining a biodegradable hydrogel with exceptional properties. Through a series of in vitro tests, GGMA/keratin hydrogels exhibited great cell compatibility via providing an appropriate environment for cell proliferation. Furthermore, this hydrogel not only exhibits extraordinary adhesive ability on visceral tissues but also extends to scenarios involving skin or organ damage, offering valuable assistance in wound healing. Our design provides a suitable platform for cell proliferation and tissue regeneration, which shows prospects for future medical research and clinical applications.
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Thrombosis, characterized by blood clot formation within vessels, poses a significant medical challenge. Despite extensive research, the development of effective thrombosis therapies is hindered by substantial costs, lengthy development times, and high failure rates in medication commercialization. Conventional pre-clinical models often oversimplify cardiovascular disease, leading to a disparity between experimental results and human physiological responses. In response, we have engineered a photothrombosis-on-a-chip system. This microfluidic model integrates human endothelium, human whole blood, and blood flow dynamics and employs the photothrombotic method. It enables precise, site-specific thrombus induction through controlled laser irradiation, effectively mimicking both normal and thrombotic physiological conditions on a single chip. Additionally, the system allows for the fine-tuning of thrombus occlusion levels via laser parameter adjustments, offering a flexible thrombus model with varying degrees of obstruction. Additionally, the formation and progression of thrombosis noted on the chip closely resemble the thrombotic conditions observed in mice in previous studies. In the experiments, we perfused recalcified whole blood with Rose Bengal into an endothelialized microchannel and initiated photothrombosis using green laser irradiation. Various imaging methods verified the model's ability to precisely control thrombus formation and occlusion levels. The effectiveness of clinical drugs, including heparin and rt-PA, was assessed, confirming the chip's potential in drug screening applications. In summary, the photothrombosis-on-a-chip system significantly advances human thrombosis modeling. Its precise control over thrombus formation, flexibility in the thrombus severity levels, and capability to simulate dual physiological states on a single platform make it an invaluable tool for targeted drug testing, furthering the development of organ-on-a-chip drug screening techniques.
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Dispositivos Laboratorio en un Chip , Trombosis , Humanos , Rayos Láser , Técnicas Analíticas Microfluídicas/instrumentación , Animales , Rosa BengalaRESUMEN
The objective of this study is to develop a device to mimic a microfluidic system of human arterial blood vessels. The device combines fluid shear stress (FSS) and cyclic stretch (CS), which are resulting from blood flow and blood pressure, respectively. The device can reveal real-time observation of dynamic morphological change of cells in different flow fields (continuous flow, reciprocating flow and pulsatile flow) and stretch. We observe the effects of FSS and CS on endothelial cells (ECs), including ECs align their cytoskeleton proteins with the fluid flow direction and paxillin redistribution to the cell periphery or the end of stress fibers. Thus, understanding the morphological and functional changes of endothelial cells on physical stimuli can help us to prevent and improve the treatment of cardiovascular diseases.
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Iliac vein compression syndrome (IVCS, or May-Thurner syndrome) occurs due to the compression of the left common iliac vein between the lumbar spine and right common iliac artery. Because most patients with compression are asymptomatic, the syndrome is difficult to diagnose based on the degree of anatomical compression. In this study, we investigated how the tilt angle of the left common iliac vein affects the flow patterns in the compressed blood vessel using three-dimensional computational fluid dynamic (CFD) simulations to determine the flow fields generated after compression sites. A patient-specific iliac venous CFD model was created to verify the boundary conditions and hemodynamic parameter set in this study. Thirty-one patient-specific CFD models with various iliac venous angles were developed using computed tomography (CT) angiograms. The angles between the right or left common iliac vein and inferior vena cava at the confluence level of the common iliac vein were defined as α1 and α2. Flow fields and vortex locations after compression were calculated and compared according to the tilt angle of the veins. Our results showed that α2 affected the incidence of flow field disturbance. At α2 angles greater than 60 degrees, the incidence rate of blood flow disturbance was 90%. In addition, when α2 and α1 + α2 angles were used as indicators, significant differences in tilt angle were found between veins with laminar, transitional, and turbulent flow (p < 0.05). Using this mathematical simulation, we concluded that the tilt angle of the left common iliac vein can be used as an auxiliary indicator to determine IVCS and its severity, and as a reference for clinical decision making.
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Due to the mortality associated with thrombosis and its high recurrence rate, there is a need to investigate antithrombotic approaches. Noninvasive site-specific thrombolysis is a current approach being used; however, its usage is characterized by the following limitations: low targeting efficiency, poor ability to penetrate clots, rapid half-life, lack of vascular restoration mechanisms, and risk of thrombus recurrence that is comparable to that of traditional pharmacological thrombolysis agents. Therefore, it is vital to develop an alternative technique that can overcome the aforementioned limitations. To this end, a cotton-ball-shaped platelet (PLT)-mimetic self-assembly framework engineered with a phototherapeutic poly(3,4-ethylenedioxythiophene) (PEDOT) platform has been developed. This platform is capable of delivering a synthetic peptide derived from hirudin P6 (P6) to thrombus lesions, forming P6@PEDOT@PLT nanomotors for noninvasive site-specific thrombolysis, effective anticoagulation, and vascular restoration. Regulated by P-selectin mediation, the P6@PEDOT@PLT nanomotors target the thrombus site and subsequently rupture under near-infrared (NIR) irradiation, achieving desirable sequential drug delivery. Furthermore, the movement ability of the P6@PEDOT@PLT nanomotors under NIR irradiation enables effective penetration deep into thrombus lesions, enhancing bioavailability. Biodistribution analyses have shown that the administered P6@PEDOT@PLT nanomotors exhibit extended circulation time and metabolic capabilities. In addition, the photothermal therapy/photoelectric therapy combination can significantly augment the effectiveness (ca. 72%) of thrombolysis. Consequently, the precisely delivered drug and the resultant phototherapeutic-driven heat-shock protein, immunomodulatory, anti-inflammatory, and inhibitory plasminogen activator inhibitor-1 (PAI-1) activities can restore vessels and effectively prevent rethrombosis. The described biomimetic P6@PEDOT@PLT nanomotors represent a promising option for improving the efficacy of antithrombotic therapy in thrombus-related illnesses.
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Trombosis , Activador de Tejido Plasminógeno , Humanos , Activador de Tejido Plasminógeno/farmacología , Biomimética , Distribución Tisular , Trombosis/tratamiento farmacológico , Terapia Trombolítica/métodosRESUMEN
Recently, studies have revealed that human herpesvirus 4 (HHV-4), also known as the Epstein-Barr virus, might be associated with the severity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Compared to SARS-CoV-2 infection alone, patients coinfected with SARS-CoV-2 and HHV-4 had higher risks of fever, inflammation, and even death, thus, confirming that HHV-4/SARS-CoV-2 coinfection in patients could benefit from clinical investigation. Although several intelligent devices can simultaneously discern multiple genes related to SARS-CoV-2, most operate via label-based detection, which restricts them from directly measuring the product. In this study, we developed a device that can replicate and detect SARS-CoV-2 and HHV-4 DNA. This device can conduct a duplex polymerase chain reaction (PCR) in a microfluidic channel and detect replicates in a non-labeled manner through a plasmonic-based sensor. Compared to traditional instruments, this device can reduce the required PCR time by 55% while yielding a similar amount of amplicon. Moreover, our device's limit of detection (LOD) reached 100 fg/mL, while prior non-labeled sensors for SARS-CoV-2 detection were in the range of ng/mL to pg/mL. Furthermore, the device can detect desired genes by extracting cells artificially infected with HHV-4/SARS-CoV-2. We expect that this device will be able to help verify HHV-4/SARS-CoV-2 coinfected patients and assist in the evaluation of practical treatment approaches.
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Thrombolytic and antithrombotic therapies are limited by short circulation time and the risk of off-target hemorrhage. Integrating a thrombus-homing strategy with photothermal therapy are proposed to address these limitations. Using glycol chitosan, polypyrrole, iron oxide and heparin, biomimicking GCPIH nanoparticles are developed for targeted thrombus delivery and thrombolysis. The nanoassembly achieves precise delivery of polypyrrole, exhibiting biocompatibility, selective accumulation at multiple thrombus sites, and enhanced thrombolysis through photothermal activation. To simulate targeted thrombolysis, a microfluidic model predicting thrombolysis dynamics in realistic pathological scenarios is designed. Human blood assessments validate the precise homing of GCPIH nanoparticles to activated thrombus microenvironments. Efficient near-infrared phototherapeutic effects are demonstrated at thrombus lesions under physiological flow conditions ex vivo. The combined investigations provide compelling evidence supporting the potential of GCPIH nanoparticles for effective thrombus therapy. The microfluidic model also offers a platform for advanced thrombolytic nanomedicine development.
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Nanopartículas , Trombosis , Humanos , Polímeros/uso terapéutico , Microfluídica , Pirroles , Fibrinolíticos/farmacología , Fibrinolíticos/uso terapéutico , Trombosis/tratamiento farmacológico , Trombosis/patología , Nanopartículas/uso terapéutico , Terapia TrombolíticaRESUMEN
A new approach to treating vascular blockages has been developed to overcome the limitations of current thrombolytic therapies. This approach involves biosafety and multimodal plasma-derived theranostic platelet vesicle incorporating iron oxide constructed nano-propellers platformed technology that possesses fluorescent and magnetic features and manifold thrombus targeting modes. The platform is capable of being guided and visualized remotely to specifically target thrombi, and it can be activated using near-infrared phototherapy along with an actuated magnet for magnetotherapy. In a murine model of thrombus lesion, this proposed multimodal approach showed an approximately 80 % reduction in thrombus residues. Moreover, the new strategy not only improves thrombolysis but also boosts the rate of lysis, making it a promising candidate for time-sensitive thrombolytic therapy.
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Within the heterogeneous tissue architecture, a comprehensive understanding of how cell shapes regulate cytoskeletal mechanics by adjusting focal adhesions (FAs) signals to correlate with the lineage commitment of mesenchymal stromal cells (MSCs) remains obscure. Here, via engineered extracellular matrices, we observed that the development of mature FAs, coupled with a symmetrical pattern of radial fiber bundles, appeared at the right-angle vertices in cells with square shape. While circular cells aligned the transverse fibers parallel to the cell edge, and moved them centripetally in a counter-clockwise direction, symmetrical bundles of radial fibers at the vertices of square cells disrupted the counter-clockwise swirling and bridged the transverse fibers to move centripetally. In square cells, the contractile force, generated by the myosin IIA-enriched transverse fibers, were concentrated and transmitted outwards along the symmetrical bundles of radial fibers, to the extracellular matrix through FAs, and thereby driving FA organization and maturation. The symmetrical radial fiber bundles concentrated the transverse fibers contractility inward to the linkage between the actin cytoskeleton and the nuclear envelope. The tauter cytoskeletal network adjusted the nuclear-actomyosin force balance to cause nuclear deformability and to increase nuclear translocation of the transcription co-activator YAP, which in turn modulated the switch in MSC commitment. Thus, FAs dynamically respond to geometric cues and remodel actin cytoskeletal network to re-distribute intracelluar tension towards the cell nucleus, and thereby controlling YAP mechanotransduction signaling in regulating MSC fate decision. STATEMENT OF SIGNIFICANCE: We decipher how cellular mechanics is self-organized depending on extracellular geometric features to correlate with mesenchymal stromal cell lineage commitment. In response to geometry constrains on cell morphology, symmetrical radial fiber bundles are assembled and clustered depending on the maturation state of focal adhesions and bridge with the transverse fibers, and thereby establishing the dynamic cytoskeletal network. Contractile force, generated by the myosin-IIA-enriched transverse fibers, is transmitted and dynamically drives the retrograde movement of the actin cytoskeletal network, which appropriately adjusts the nuclear-actomyosin force balance and deforms the cell nucleus for YAP mechano-transduction signaling in regulating mesenchymal stromal cell fate decision.
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Actinas , Células Madre Mesenquimatosas , Actinas/metabolismo , Actomiosina/metabolismo , Mecanotransducción Celular , Forma de la Célula , Osteogénesis , Diferenciación Celular , Factores de Transcripción/metabolismoRESUMEN
Traditional thrombolytic therapeutics for vascular blockage are affected by their limited penetration into thrombi, associated off-target side effects, and low bioavailability, leading to insufficient thrombolytic efficacy. It is hypothesized that these limitations can be overcome by the precisely controlled and targeted delivery of thrombolytic therapeutics. A theranostic platform is developed that is biocompatible, fluorescent, magnetic, and well-characterized, with multiple targeting modes. This multimodal theranostic system can be remotely visualized and magnetically guided toward thrombi, noninvasively irradiated by near-infrared (NIR) phototherapies, and remotely activated by actuated magnets for additional mechanical therapy. Magnetic guidance can also improve the penetration of nanomedicines into thrombi. In a mouse model of thrombosis, the thrombosis residues are reduced by ≈80% and with no risk of side effects or of secondary embolization. This strategy not only enables the progression of thrombolysis but also accelerates the lysis rate, thereby facilitating its prospective use in time-critical thrombolytic treatment.
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Terapia Trombolítica , Trombosis , Ratones , Animales , Medicina de Precisión , Fibrinolíticos/química , Fibrinolíticos/uso terapéutico , Trombosis/diagnóstico por imagen , Trombosis/tratamiento farmacológico , Fenómenos MagnéticosRESUMEN
In the fields of biomedicine and tissue engineering, natural polymer-based tissue-engineered scaffolds are used in multiple applications. As a plant-derived polymer, soy protein, containing multiple amino acids, is structurally similar to components of the extra-cellular matrix (ECM) of tissues. It is biological safety provided a good potential to be material for pure natural scaffolds. Moreover, as a protein, the properties of soy protein can be easily adjusted by modifying the functional groups on it. In addition, by blending soy protein with other synthetic and natural polymers, the mechanical characteristics and bioactive behavior of scaffolds can be facilitated for a variety of bio-applications. In this research, soy protein and polysaccharides tapioca starch are used, and gellan gum to develop a protein-based composite scaffold for cell engineering. The morphology and surface chemical composition are characterized via micro-computed tomography (micro-CT), scanning electron microscope (SEM), and fourier-transform infrared (FTIR) spectroscopy. The soy/tapioca/gellan gum (STG) composite scaffolds selectively help the adhesion and proliferation of L929 fibroblast cells while improving the migration of L929 fibroblast cells in STG composite scaffolds as the increase of soy protein proportion of the scaffold. In addition, STG composite scaffolds show great potential in the wound healing model to enhance rapid epithelialization and tissue granulation.
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Manihot , Proteínas de Soja , Proteínas de Soja/farmacología , Proteínas de Soja/química , Microtomografía por Rayos X , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Vendajes , Polímeros/química , Almidón/farmacologíaRESUMEN
Noninvasive photothermal therapy (PTT) represents a promising direction for more modern and precise medical applications. However, PTT efficacy is still not satisfactory due to the existence of heat shock proteins (HSPs) and poorly targeted delivery. Herein, the design of a nanosystem with improved delivery efficacy for anticancer treatment employing the synergetic effects of reactive oxygen species (ROS)-driven chemodynamic therapy (CDT) to inactivated HSPs with photothermal-hyperthermia was therefore achieved through the development of pH-targeting glycol chitosan/iron oxide enclosed core polypyrrole nanoclusters (GCPI NCs). The designed NCs effectively accumulated toward cancer cells due to their acidic microenvironment, initiating ROS generation via Fenton reaction at the outset and performing site-specific near infrared (NIR)-photothermal effect. A comprehensive analysis of both surface and bulk material properties of the CDT/PTT NCs as well as biointerface properties were ascertained via numerous surface specific analytical techniques by bringing together heightened accumulation of CDT/PTT NCs, which can significantly eradicate cancer cells thus minimizing the side effects of conventional chemotherapies. All of these attributes act in synergy over the cancer cells succeeding in fashioning NC's able to act as competent agents in the MRI-monitored enhanced CDT/PTT synergistic therapy. Findings in this study evoke attention in future oncological therapeutic strategies.
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Terapia Fototérmica , Polímeros , Línea Celular Tumoral , Quitosano , Compuestos Férricos , Pirroles/farmacologíaRESUMEN
This paper reports a novel micro/nanostructure co-hot embossing technique. Gold-capped nanostructures were used as localized surface plasmon resonance (SPR) sensors and were integrated into a microfluidic channel. The advantage of the co-hot embossing technique is that the SPR sensors do not need to be aligned with the microfluidic channel while bonding to it. The integrated SPR sensor and microfluidic channel were first characterized, and the sensitivity of the SPR sensor to the refractive index was found using different concentrations of glycerol solutions. The SPR sensor was also used to quantify latent membrane protein (LMP-1) when modifying anti-LMP-1 at the surface of the SPR sensor. Different concentrations of LMP-1 samples were used to build a calibration curve.
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Técnicas Biosensibles , Resonancia por Plasmón de Superficie , Técnicas Biosensibles/métodos , Oro/química , Dispositivos Laboratorio en un Chip , Proteínas de la Membrana , Resonancia por Plasmón de Superficie/métodosRESUMEN
Surface topography-induced lineage commitment of human bone marrow stem cells (hBMSCs) has been reported. However, this effect on hBMSC differentiation toward retinal pigment epithelium (RPE)-like cells has not been explored. Herein, a family of cell culture substrates called binary colloidal crystals (BCCs) was used to stimulate hBMSCs into RPE-like cells without induction factors. Two BCCs, named SiPS (silica (Si)/polystyrene (PS)) and SiPSC (Si/carboxylated PS), having similar surface topographies but different surface chemistry was used for cell culture. The result showed that cell proliferation was no difference between the two BCCs and tissue culture polystyrene (TCPS) control. However, the cell attachment, spreading area, and aspect ratio between surfaces were significantly changed. For example, cells displayed more elongated on SiPS (aspect ratio ~7.0) than those on SiPSC and TCPS (~2.0). The size of focal adhesions on SiPSC (~1.6 µm2) was smaller than that on the TCPS (~2.5 µm2). qPCR results showed that hBMSCs expressed higher RPE progenitor genes (i.e., MITF and PAX6) on day 15, and mature RPE genes (i.e., CRALBP and RPE65) on day 30 on SiPS than TCPS. On the other hand, the expression of optical vesicle or neuroretina genes (i.e., MITF and VSX2) was upregulated on day 15 on SiPSC compared to the TCPS. This study reveals that hBMSCs could be modulated into different cell subtypes depending on the BCC combinations. This study shows the potential of BCCs in controlling stem cell differentiation.
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Poliestirenos , Retina , Humanos , Diferenciación Celular , Expresión Génica , Poliestirenos/farmacología , Dióxido de Silicio/farmacologíaRESUMEN
In this study, microneedle-integrated light sheet microscopy (LSM) was developed for trapping and continuously imaging embryos of Caenorhabditis elegans with subcellular resolution. To reduce aberrations when the light sheet was propagated into the device, a microneedle was fabricated using a transparent, water refractive index-matched polymer. It was proven that when the light sheet emerged from the water-immersed objective and penetrated through the microneedle with a circular surface, even with a non-perpendicular incident angle, fewer aberrations were found. An embryo was injected into and trapped at the tip of the microneedle, which was positioned at the interrogation window of the LSM apparatus with the image plane perpendicular to the light sheet, and this setup was used to sequentially acquire embryo images. By applying the light sheet, higher-resolution, higher-contrast images were obtained. The system also showed low photobleaching and low phototoxicity to embryos of C. elegans. Furthermore, three-dimensional embryo images with a whole field of view of the microneedle could be achieved by stitching together images and reconstructing sequential two-dimensional embryo images.
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Microscopía , Refractometría , Animales , Caenorhabditis elegans , Microscopía/métodos , Fotoblanqueo , AguaRESUMEN
We present the first combination of a microfluidic polymerase chain reaction (PCR) with a gold nanoslit-based surface plasmon resonance (SPR) sensor for detecting the DNA sequence of latent membrane protein 1 (LMP1). The PCR microchannel was produced through a laser scribing technique, and the SPR nanoslit chip was manufactured via hot-embossing nanoimprinting lithography. Afterward, the LMP1 DNA probe was adsorbed onto the SPR chip of the integrated device through electrostatic interactions for further detection. The device can complete the analytical procedure in around 36 min, while the traditional machine requires 105 min to achieve similar signals under the same PCR thermal cycles. The calibration curve with serially diluted LMP1 DNA exhibited the accuracy (R2 > 0.99) and sensitivity (limit of detection: â¼10-11 g/mL) of the device. Moreover, extracted DNA from Epstein-Barr virus (EBV)-positive cells were directly detected through the integrated chip. In brief, this all-in-one chip can amplify gene fragments at the front-end and detect them at the back-end, decreasing the time required for the analysis without compromising accuracy or sensitivity. We believe this label-free, real-time, low-cost device has enormous potential for rapid detection of various viruses, such as EBV and COVID-19.
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Técnicas Biosensibles , COVID-19 , Infecciones por Virus de Epstein-Barr , Oro , Herpesvirus Humano 4/genética , Humanos , Microfluídica , Reacción en Cadena de la Polimerasa , SARS-CoV-2 , Proteínas de la Matriz Viral/genéticaRESUMEN
Rheumatoid arthritis (RA), an autoimmune and chronic inflammatory disorder, is an incurable disease. We developed a peptide-based electrochemical sensor using electrochemical impedance spectroscopy that can be used to detect autoantibodies for RA diagnostics. We first validated that the developed peptide showed high sensitivity and could compliment the current gold standard method of an anti-cyclic citrullinated peptide antibody (anti-CCP) ELISA. The developed peptide can be modified on the nanogold surface of the working electrode of sensing chips through the method of a self-assembling monolayer. The sensing process was first optimized using a positive control cohort and a healthy control cohort. Subsequently, 10 clinically confirmed samples from RA patients and five healthy control samples were used to find the threshold value of the impedance between RA and healthy subjects. Furthermore, 10 clinically confirmed samples but with low values of anti-CCP autoantibodies were used to evaluate the sensitivity of the present method compared to the conventional method. The proposed method showed better sensitivity than the current conventional anti-CCP ELISA method.