Aptamer-Based Smart Targeting and Spatial Trigger-Response Drug-Delivery Systems for Anticancer Therapy.

In recent years, the field of drug delivery has witnessed remarkable progress, driven by the quest for more effective and precise therapeutic interventions. Among the myriad strategies employed, the integration of aptamers as targeting moieties and stimuli-responsive systems has emerged as a promising avenue, particularly in the context of anticancer therapy. This review explores cutting-edge advancements in targeted drug-delivery systems, focusing on the integration of aptamers and stimuli-responsive platforms for enhanced spatial anticancer therapy. In the aptamer-based drug-delivery systems, we delve into the versatile applications of aptamers, examining their conjugation with gold, silica, and carbon materials. The synergistic interplay between aptamers and these materials is discussed, emphasizing their potential in achieving precise and targeted drug delivery. Additionally, we explore stimuli-responsive drug-delivery systems with an emphasis on spatial anticancer therapy. Tumor microenvironment-responsive nanoparticles are elucidated, and their capacity to exploit the dynamic conditions within cancerous tissues for controlled drug release is detailed. External stimuli-responsive strategies, including ultrasound-mediated, photo-responsive, and magnetic-guided drug-delivery systems, are examined for their role in achieving synergistic anticancer effects. This review integrates diverse approaches in the quest for precision medicine, showcasing the potential of aptamers and stimuli-responsive systems to revolutionize drug-delivery strategies for enhanced anticancer therapy.

Therapeutic Agent-Loaded Fibrous Scaffolds for Biomedical Applications.

Tissue engineering is a sophisticated field that involves the integration of various disciplines, such as clinical medicine, material science, and life science, to repair or regenerate damaged tissues and organs. To achieve the successful regeneration of damaged or diseased tissues, it is necessary to fabricate biomimetic scaffolds that provide structural support to the surrounding cells and tissues. Fibrous scaffolds loaded with therapeutic agents have shown considerable potential in tissue engineering. In this comprehensive review, we examine various methods for fabricating bioactive molecule-loaded fibrous scaffolds, including preparation methods for fibrous scaffolds and drug-loading techniques. Additionally, we delved into the recent biomedical applications of these scaffolds, such as tissue regeneration, inhibition of tumor recurrence, and immunomodulation. The aim of this review is to discuss the latest research trends in fibrous scaffold manufacturing methods, materials, drug-loading methods with parameter information, and therapeutic applications with the goal of contributing to the development of new technologies or improvements to existing ones.

Design and Prediction of Aptamers Assisted by In Silico Methods.

An aptamer is a single-stranded DNA or RNA that binds to a specific target with high binding affinity. Aptamers are developed through the process of systematic evolution of ligands by exponential enrichment (SELEX), which is repeated to increase the binding power and specificity. However, the SELEX process is time-consuming, and the characterization of aptamer candidates selected through it requires additional effort. Here, we describe in silico methods in order to suggest the most efficient way to develop aptamers and minimize the laborious effort required to screen and optimise aptamers. We investigated several methods for the estimation of aptamer-target molecule binding through conformational structure prediction, molecular docking, and molecular dynamic simulation. In addition, examples of machine learning and deep learning technologies used to predict the binding of targets and ligands in the development of new drugs are introduced. This review will be helpful in the development and application of in silico aptamer screening and characterization.

Principles and Applications of Loop-Mediated Isothermal Amplification to Point-of-Care Tests.

For the identification of nucleic acids, which are important biomarkers of pathogen-mediated diseases and viruses, the gold standard for NA-based diagnostic applications is polymerase chain reaction (PCR). However, the requirements of PCR limit its application as a rapid point-of-care diagnostic technique. To address the challenges associated with regular PCR, many isothermal amplification methods have been developed to accurately detect NAs. Isothermal amplification methods enable NA amplification without changes in temperature with simple devices, as well as faster amplification times compared with regular PCR. Of the isothermal amplifications, loop-mediated isothermal amplification (LAMP) is the most studied because it amplifies NAs rapidly and specifically. This review describes the principles of LAMP, the methods used to monitor the process of LAMP, and examples of biosensors that detect the amplicons of LAMP. In addition, current trends in the application of LAMP to smartphones and self-diagnosis systems for point-of-care tests are also discussed.

Wearable Sensing Devices: Towards the Development of a Personalized System for Construction Safety and Health Risk Mitigation.

Wearable sensing devices (WSDs) are increasingly helping workers stay safe and healthy in several industries. However, workers, especially in the construction industry, have shown some aversion towards the use of WSDs due to their ability to capture specific information that may be considered personal and private. However, this revered information may provide some critical insight needed by management to plan and optimize worksite safety and support technology adoption in decision making. Therefore, there is a need to develop personalized WSD systems that are mutually beneficial to workers and management to ensure successful WSD integration. The present study aims to contribute to knowledge and practice by filling this critical gap using insight from 330 construction workers with experience using WSDs. The results from this study indicate that all 11 WSD functions identified through this study play a vital role in improving worker safety and health and that approximately two out of three workers are open to sharing the physiological and environmental information captured using these WSDs with their management. However, functions for detecting workers' proximity to workplace hazards, specifically energized electrical materials, toxic gas, and fire/smoke, were the most critical functions that had mutual value to workers and management. Finally, the present study proposed and evaluated a phased personalized WSD system that should encourage successful WSD integration.

Multi-Level-Phase Deep Learning Using Divide-and-Conquer for Scaffolding Safety.

A traditional structural analysis of scaffolding structures requires loading conditions that are only possible during design, but not in operation. Thus, this study proposes a method that can be used during operation to make an automated safety prediction for scaffolds. It implements a divide-and-conquer technique with deep learning. As a test scaffolding, a four-bay, three-story scaffold model was used. Analysis of the model led to 1411 unique safety cases for the model. To apply deep learning, a test simulation generated 1,540,000 datasets for pre-training, and an additional 141,100 datasets for testing purposes. The cases were then sub-divided into 18 categories based on failure modes at both global and local levels, along with a combination of member failures. Accordingly, the divide-and-conquer technique was applied to the 18 categories, each of which were pre-trained by a neural network. For the test datasets, the overall accuracy was 99%. The prediction model showed that 82.78% of the 1411 safety cases showed 100% accuracy for the test datasets, which contributed to the high accuracy. In addition, the higher values of precision, recall, and F1 score for the majority of the safety cases indicate good performance of the model, and a significant improvement compared with past research conducted on simpler cases. Specifically, the method demonstrated improved performance with respect to accuracy and the number of classifications. Thus, the results suggest that the methodology could be reliably applied for the safety assessment of scaffolding systems that are more complex than systems tested in past studies. Furthermore, the implemented methodology can easily be replicated for other classification problems.

High-throughput label-free molecular fingerprinting flow cytometry.

Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm-1) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics of Haematococcus lacustris.

Microfluidic single-particle chemical analyzer with dual-comb coherent Raman spectroscopy.

Label-free particle analysis is a powerful tool in chemical, pharmaceutical, and cosmetic industries as well as in basic sciences, but its throughput is significantly lower than that of fluorescence-based counterparts. Here we present a label-free single-particle analyzer based on broadband dual-comb coherent Raman scattering spectroscopy operating at a spectroscopic scan rate of 10 kHz. As a proof-of-concept demonstration, we perform broadband coherent anti-Stokes Raman scattering measurements of polystyrene microparticles flowing on an acoustofluidic chip at a high throughput of >1000 particles per second. This high-throughput label-free particle analyzer has the potential for high-precision statistical analysis of a large number of microparticles including biological cells.

Acoustofluidic harvesting of microalgae on a single chip.

We present an on-chip acoustofluidic platform for harvesting a target microalgal species from a heterogeneous population of cells and particles based on their size, density, and compressibility in a rapid, non-invasive, energy-efficient, continuously running, and automated manner. For our proof-of-principle demonstration, we use Euglena gracilis as a target species. Specifically, we show the simultaneous separation and enrichment of E. gracilis from a mixed population of E. gracilis in pond water (consisting of other microalgae and various kinds of particles as contaminants) on a single acoustofluidic chip with a recovery ratio of 92.6%, a target separation ratio of 90.1%, a concentration factor of 3.43, an enrichment factor of 12.76, and a cell viability rate of 98.3% at a high volume rate of 500 µl/min. Our results indicate that the on-chip acoustofluidic platform is an effective tool for harvesting target microalgae from mixed populations of microalgae and other contaminants.

Acousto-microfluidics for screening of ssDNA aptamer.

We demonstrate a new screening method for obtaining a prostate-specific antigen (PSA) binding aptamer based on an acoustofluidic separation (acoustophoreis) technique. Since acoustophoresis provides simultaneous washing and separation in a continuous flow mode, we efficiently obtained a PSA binding aptamer that shows high affinity without any additional washing step, which is necessary in other screening methods. In addition, next-generation sequencing (NGS) was applied to accelerate the identification of the screened ssDNA pool, improving the selecting process of the aptamer candidate based on the frequency ranking of the sequences. After the 8(th) round of the acoustophoretic systematic evolution of ligands by exponential enrichment (SELEX) and following sequence analysis with NGS, 7 PSA binding ssDNA aptamer-candidates were obtained and characterized with surface plasmon resonance (SPR) for affinity and specificity. As a result of the new SELEX method with PSA as the model target protein, the best PSA binding aptamer showed specific binding to PSA with a dissociation constant (Kd) of 0.7 nM.

Aptamer/ISET-MS: a new affinity-based MALDI technique for improved detection of biomarkers.

With the rapid progress in the development of new clinical biomarkers there is an unmet need of fast and sensitive multiplex analysis methods for disease specific protein monitoring. Immunoaffinity extraction integrated with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis offers a route to rapid and sensitive protein analysis and potentially multiplex biomarker analysis. In this study, the previously reported integrated selective enrichment target (ISET)-MALDI-MS analysis was implemented with ssDNA aptamer functionalized microbeads to address the specific capturing of thrombin in complex samples. The main objective for using an aptamer as the capturing ligand was to avoid the inherently high background components, which are produced during the digestion step following the target extraction when antibodies are used. By applying a thrombin specific aptamer linked to ISET-MALDI-MS detection, a proof of concept of antibody fragment background reduction in the ISET-MALDI-MS readout is presented. Detection sensitivity was significantly increased compared to the corresponding system based on antibody-specific binding as the aptamer ligand does not induce any interfering background residues from the antibodies. The limit of detection for thrombin was 10 fmol in buffer using the aptamer/ISET-MALDI-MS configuration as confirmed by MS/MS fragmentation. The aptamer/ISET-MALDI-MS platform also displayed a limit of detection of 10 fmol for thrombin in five different human serum samples (1/10 diluted), demonstrating the applicability of the aptamer/ISET-MALDI-MS analysis in clinical samples.

An ultra-sensitive detection of a whole virus using dual aptamers developed by immobilization-free screening.

In this study, we successfully developed a ssDNA aptamer pairs by using an advanced immobilization-free SELEX method with affinity-based selection and counter-screening process at every round. By implementing this method, two different aptamers specifically binding to bovine viral diarrhea virus type 1(BVDV type 1) with high affinity were successfully screened. This aptamer pair was applied to ultrasensitive detection platform for BVDV type 1 in a sandwich manner. The ultrasensitive detection of BVDV type 1 using one of aptamers conjugated with gold nanoparticles was obtained in aptamer-aptamer sandwich type sensing format, with the limit of detection of 800 copies/ml, which is comparable to a real-time PCR method.

Rapid and sensitive detection of Nampt (PBEF/visfatin) in human serum using an ssDNA aptamer-based capacitive biosensor.

A single-stranded DNA (ssDNA) aptamer was successfully developed to specifically bind to nicotinamide phosphoribosyl transferase (Nampt) through systematic evolution of ligands by exponential enrichment (SELEX) and successfully implemented in a gold-interdigitated (GID) capacitor-based biosensor. Surface plasmon resonance (SPR) analysis of the aptamer revealed high specificity and affinity (K(d)=72.52 nM). Changes in surface capacitance/charge distribution or dielectric properties in the response of the GID capacitor surface covalently coupled to the aptamers in response to changes in applied AC frequency were measured as a sensing signal based on a specific interaction between the aptamers and Nampt. The limit of detection for Nampt was 1 ng/ml with a dynamic serum detection range of up to 50 ng/ml; this range includes the clinical requirement for both normal Nampt level, which is 15.8 ng/ml, and Nampt level in type 2 diabetes mellitus (T2DM) patients, which is 31.9 ng/ml. Additionally, the binding kinetics of aptamer-Nampt interactions on the capacitor surface showed that strong binding occurred with increasing frequency (range, 700 MHz-1 GHz) and that the dissociation constant of the aptamer under the applied frequency was improved 120-240 times (K(d)=0.3-0.6 nM) independent on frequency. This assay system is an alternative approach for clinical detection of Nampt with improved specificity and affinity.

Immobilization-free screening of aptamers assisted by graphene oxide.

Graphene oxide (GO) has the ability to separate free short ssDNA in heterogeneous solution. This feature is applied as a label free platform for screening of aptamers that bind to their target with high affinity and specificity. Herein, we report an aptamer selection strategy for Nampt protein based on GO.