Nanoarchitectonics-based electrochemical aptasensors for highly efficient exosome detection.

Exosomes, a type of extracellular vesicles, have attracted considerable attention due to their ability to provide valuable insights into the pathophysiological microenvironment of the cells from which they originate. This characteristic implicates their potential use as diagnostic disease biomarkers clinically, including cancer, infectious diseases, neurodegenerative disorders, and cardiovascular diseases. Aptasensors, which are electrochemical aptamers based biosensing devices, have emerged as a new class of powerful detection technology to conventional methods like ELISA and Western analysis, primarily because of their capability for high-performance bioanalysis. This review covers the current research landscape on the detection of exosomes utilizing nanoarchitectonics strategy for the development of electrochemical aptasensors. Strategies involving signal amplification and biofouling prevention are discussed, with an emphasis on nanoarchitectonics-based bio-interfaces, showcasing their potential to enhance sensitivity and selectivity through optimal conduction and mass transport properties. The ongoing challenges to broaden the clinical applications of these biosensors are also highlighted.

This review emphasizes the significant impact of integrating nanoarchitectonics into aptamer-based electrochemical biosensors for exosome detection, thereby enhancing early disease detection and monitoring disease progression in clinical settings.

Enhancing Substrate Channeling with Multi-Enzyme Architectures in Hydrogen-Bonded Organic Framework.

Hydrogen-bonded organic frameworks (HOF) represent an emerging category of organic structures with high crystallinity and metal-free, which are not commonly observed in alternative porous organic frameworks. These needle-like porous structure can help in stabilizing enzymes and allow transfer of molecules between enzymes participating in cascade reactions for enhanced substrate channelling. Herein, we systematically synthesized and investigated the stability of HOF at extreme conditions followed by one-pot encapsulation of single and bi-enzyme systems. Firstly, we observed HOF to be stable at pH 1 to 14 and at high temperatures (up to 115°C). Secondly, the encapsulated glucose oxidase enzyme (GOX) showed 80 % and 90% of its original activity at 70 °C and pH 11, respectively. Thirdly, transient time close to 0 seconds was observed for HOF encapsulated bi-enzyme cascade reaction system demonstrating a 4.25-fold improvement in catalytic activity when compared to free enzymes with enhanced substrate channelling. Our findings showcase a facile system synthesized under ambient conditions to encapsulate and stabilize enzymes at extreme conditions.

In Situ Immobilization of Multi-Enzymes for Enhanced Substrate Channeling of Enzyme Cascade Reactions: A Nanoarchitectonics Approach by Directed Metal-Organic Frameworks.

Rationally tailoring a controlled spatial organization of enzymes in a nanoarchitecture for multi-enzyme cascade reactions can enhance the catalytic efficiency via substrate channeling. However, attaining substrate channeling is a grand challenge, requiring sophisticated techniques. Herein, we report facile polymer-directed metal-organic framework (MOF)-based nanoarchitechtonics for realizing a desirable enzyme architecture with significantly enhanced substrate channeling. The new method involves the use of poly(acrylamide-co-diallyldimethylammonium chloride) (PADD) as a modulator in a one-step process for simultaneous MOF synthesis and co-immobilization of enzymes (GOx and HRP). The resultant enzymes-PADD@MOFs constructs showed a closely packed nanoarchitecture with enhanced substrate channeling. A transient time close to 0 s was observed, owing to a short diffusion path for substrates in a 2D spindle-shaped structure and their direct transfer from one enzyme to another. This enzyme cascade reaction system showed a 3.5-fold increase in catalytic activity in comparison to free enzymes. The findings provide a new insight into using polymer-directed MOF-based enzyme nanoarchitectures to improve catalytic efficiency and selectivity.

Nucleic acid isothermal amplification-based soft nanoarchitectonics as an emerging electrochemical biosensing platform.

The emergence of nucleic acid isothermal amplification strategies based on soft nanoarchitectonics offers a new dimension to the traditional electrochemical technique, particularly because of its flexibility, high efficiency, and increased sensitivity for analytical applications. Various DNA/RNA isothermal amplification strategies have been developed for the design and fabrication of new electrochemical biosensors for efficient and important biomolecular detection. Herein, we provide an overview of recent efforts in this research field and the strategies for signal-amplified sensing systems, with their biological applications, current challenges and prospects in this promising new area.

2D Active Nanobots Based on Soft Nanoarchitectonics Powered by an Ultralow Fuel Concentration.

Enzyme catalysis to power micro/nanomotors has received tremendous attention because of the vast potential in applications ranging from biomedicine to environmental remediation. However, the current design is mainly based on a complex three-dimensional (3D) architecture, with limited accessible surface areas for the catalytic sites, and thus requires a higher fuel concentration to achieve active motion. Herein we report for the first time an enzyme-powered 2D nanobot, which was designed by a facile strategy based on soft nanoarchitectonics for active motion at an ultralow fuel concentration (0.003 % H2 O2 ). The 2D nanobots exhibited efficient positive chemotactic behavior and the ability to swim against gravity by virtue of solutal buoyancy. As a proof-of-concept, the 2D nanobots showed an excellent capability for "on-the-fly" removal of methylene blue (MB) dye with an efficiency of 85 %.

Mechanochemistry: A force in disguise and conditional effects towards chemical reactions.

Mechanochemistry refers to unusual chemical reactions induced by mechanical energy at room temperatures. It has attracted increased attention because of advantages, such as being a solution-free, energy saving, high-productivity and low-temperature process. However, there is limited understanding of the mechanochemical process because mechanochemistry is often conducted using closed milling devices, which are often regarded as a black box. This feature article shows that mechanochemical reactions can be controlled by varying milling parameters, such as the mechanical force, milling intensity, time and atmosphere. New nanomaterials with doped and functionalized structures can be produced under controlled conditions, which provide a critical insight for understanding mechanochemistry. A fundamental mechanism investigation using force microscopy is discussed.

Supramolecular nanomotors with "pH taxis" for active drug delivery in the tumor microenvironment.

Self-propelled nanomotors demonstrating autonomous motion in biologically relevant fuel are currently being studied to overcome the use of external physical or chemical stimuli as precise delivery agents. In this context, the tumor microenvironment (TME) with slightly acidic pH is used for developing cargo-releasing artificial systems triggered by such conditions. However, there is still a need for fabrication of smart nanomotors that can sense the acidic pH prevalent in the TME rather than using an external fuel source for selective activation and thereafter migrating towards tumors for active drug delivery. Herein, supramolecular assembly-based nanomotors are fabricated by in-situ grown CaCO3 nanoparticles and studied for their motility behaviour in endogenously generated acidic pH by HeLa cells and further exploited as an active delivery vehicle for DOX molecules to the cells for their anticancer efficacy. The nanomotors are activated in slightly acidic pH showcasing "pH taxis" towards tumor cells without the need for any sophisticated/complicated technologies or an external fuel source for active and targeted delivery of drugs.

Solvent Effect on Supramolecular Self-Assembly of Chlorophylls a on Chemically Reduced Graphene Oxide.

Solvent plays an important role in the surface interaction of molecules. In this study, we use "chlorophyll a", an archetypical molecule, to investigate its supramolecular self-assembly with chemically reduced graphene oxide in three different types of solvents: polar protic, polar aprotic, and non-polar. It was observed that only a polar protic solvent that can donate protons facilitates the hydrogen bonding between chlorophyll a and chemically reduced graphene oxide nanosheets in a hybrid system. The formation of hydrogen bonds further initiates the other non-covalent interactions such as π-π stacking and hydrophobic interaction, which altogether play a key driving force for supramolecular self-assembly of chlorophylls on chemically reduced graphene oxides. The experimental results are strongly supported by density functional theory calculations, which show robust electron coupling between chlorophylls and chemically reduced graphene oxide.

Enzyme catalysis powered micro/nanomotors for biomedical applications.

With recent developments in the field of autonomous motion for artificial systems, many researchers are focusing on their biomedical application for active and targeted delivery. In this context, enzyme powered motors are at the forefront since they can utilize physiologically relevant fuels as their substrate and carry out catalytic reactions to power motion under in vivo conditions. This review focuses on the design and fabrication of enzyme powered motors together with their propulsion mechanism by using fuels present in biological environments. In addition, the recent advances in the field of enzyme powered motors for biomedical applications have been discussed together with the parameters that need to be considered for designing such systems. We believe that this review will provide insights and better understanding for the development of next generation biomedical technologies based on enzyme powered motors.

Enzyme-Powered Nanomotors with Controlled Size for Biomedical Applications.

Self-propelled motors have been developed with promising potential for medical applications. However, most of them have a size range at the microscale, which limits their further research for in vivo experiments. Previously, our group developed nanoscaled motors with a size of around 400 nm with several merits, for example, delivering both hydrophobic and hydrophilic drugs/proteins, using biocompatible fuels while being able to control their motion, and showing adaptive changes of their speed and navigation to changes in the environment. It is also well-known that nanoparticles that are around 20-200 nm in size have advantages in overcoming cellular barriers and being internalized into cells. Therefore, lowering the size range of this stomatocyte nanomotor is crucial. However, the strict control of the size of vesicles in such a low regime as well as their shape transformation into folded stomatocyte structures is not trivial. In this study, we fabricated ultrasmall stomatocyte polymersomes with the size of around 150 nm, which could be a promising carrier for biomedical purposes. We demonstrated that the addition of PEG additive allows for both shape transformation of small polymersomes into stomatocytes and encapsulation of biologics. Biocatalyst catalase was encapsulated in the inner compartment of the nanomotor, protecting the enzyme while providing enough thrust to propel the motors. The ultrasmall stomatocyte motor system allowed propelled motion by converting H2O2 into O2 in the presence of only 2 mM H2O2, and the velocity of motors correlated to the O2 production. Compared to small stomatocyte nanomotors, ultrasmall stomatocyte motors demonstrate enhanced penetration across the vasculature model and increased uptake by HeLa cells in the presence of fuel.

Improved Corrosion Resistance of Steel in Ethanol Fuel Blend by Titania Nanoparticles and Aganonerion polymorphum Leaf Extract.

A porous and low-density protective film on a steel surface in the corrosive environment can undergo deterioration even in the presence of organic inhibitors due to infiltration of aggressive ions into the pinholes and/or pores. This phenomenon is related to the localized corrosion that takes place even in the presence of an optimal concentration of organic corrosion inhibitors in the given medium. To overcome this issue, we have designed an organic protective film on a steel surface with the help of titania nanoparticles (TNPs) combined with an organic corrosion inhibitor derived from Aganonerion polymorphum leaf extract (APLE), all to be studied in a simulated ethanol fuel blend (SEFB). The TNPs with varied diameters and concentrations have been studied for examining their effect on the inhibition capacity of 1000 ppm APLE on the steel surface in SEFB medium using electrochemical and surface analysis techniques. Enhanced corrosion inhibition of the surficial film was observed in the presence of both the APLE inhibitor and small amounts of TNPs. A direct agreement was observed between the experimental and molecular dynamics theoretical investigations showcasing high binding energy between inhibitor molecules and steel substrates, resulting in a much higher adhesion of the protective film, good thermal stability of the adsorbent film, and electron abundance for the supply of steel substrate of inhibitor species.

A Supramolecular Approach to Nanoscale Motion: Polymersome-Based Self-Propelled Nanomotors.

Autonomous micro- and nanoscale systems have revolutionized the way scientists look into the future, opening up new frontiers to approach and solve problems via a more bioinspired route. However, to achieve systems with higher complexity, superior output control, and multifunctionality, an in-depth study of the different factors that affect micro- and nanomotor behavior is crucial. From a fundamental perspective, the mechanical response of micro- and nanomotors still requires further study in order to have a better understanding of how exactly these systems operate and the different mechanisms of motion that can be combined into one system to achieve an optimal response. From a design engineering point of view, compatibility, degradability, specificity, sensitivity, responsiveness, and efficiency of the active systems fabricated to this point have to be addressed, with respect to the potential of these devices for biomedical applications. Nonetheless, optimizing the system with regards to all these areas is a challenging task with the micro- and nanomotors studied to date, as most of them consist of materials or designs that are unfavorable for further chemical or physical manipulation. As this new field of self-powered systems moves forward, the need for motor prototypes with different sizes, shapes, chemical functionalities, and architectures becomes increasingly important and will define not only the way active systems are powered, but also the methods for motor fabrication. Bottom-up supramolecular approaches have recently emerged as great candidates for the development of active structures that allow for chemical or physical functionalization, shape transformation, and compartmentalization, in a structure that provides a soft interface to improve molecular recognition and cell uptake. Our group pioneers the use of supramolecular structures as catalytically propelled systems via the fabrication of stomatocyte or tubular-shaped motors capable of displaying active motion in a substrate concentration-dependent fashion. This behavior demonstrates the potential of bottom-up assemblies for powering motion at the micro- or nanoscale, with a system that can be readily tuned and controlled at the molecular level. In this Account, we highlight the steps we have taken in order to understand and optimize the design of catalytically powered polymersome-based motors. Our research has been focused on addressing the importance of motor architecture, motion activation, direction control, and biological integration. While our work supports the feasibility of supramolecular structures for the design of active systems, we strongly believe that we are still in the initial stages of unveiling the full potential of supramolecular chemistry in the micro- and nanomotor field. We look forward to using this approach for the development of multifunctional and stimuli-responsive systems in the near future.

Transforming doxorubicin into a cancer stem cell killer via EpCAM aptamer-mediated delivery.

Chemotherapy-resistant cancer stem cells (CSCs) are a major obstacle to the effective treatment of many forms of cancer. To overcome CSC chemo-resistance, we developed a novel system by conjugating a CSC-targeting EpCAM aptamer with doxorubicin (Apt-DOX) to eliminate CSCs. Incubation of Apt-DOX with colorectal cancer cells resulted in high concentration and prolonged retention of DOX in the nuclei. Treatment of tumour-bearing xenograft mice with Apt-DOX resulted in at least 3-fold more inhibition of tumour growth and longer survival as well as a 30-fold lower frequency of CSC and a prolonged longer tumourigenic latency compared with those receiving the same dose of free DOX. Our data demonstrate that a CSC-targeting aptamer is able to transform a conventional chemotherapeutic agent into a CSC-killer to overcome drug resistance in solid tumours.

Graphene-Oxide-Based Enzyme Nanoarchitectonics for Substrate Channeling.

The controlled spatial organization or compartmentalization of multi-enzyme cascade reactions to transfer a substrate from one enzyme to another for substrate channeling on scaffolds has sparked increasing interest in recent years. Here, we use graphene oxides to study the dependence of the activity of cascade reactions in a closely packed, randomly immobilized enzyme system on a 2 D scaffold. We first observe that the hydrophobicity of graphene oxides and various enzyme architectures for co-immobilized systems are important attributes for achieving high product-conversion rates. A transient time close to 0 s can be achieved if enzymes are randomly immobilized close to one another, owing to direct molecular channeling. This contributes to overcoming complications regarding control of the spatial arrangement of the enzymes. Furthermore, a fabricated bienzyme paper can be used for glucose detection with high stability, reusability, and enhanced substrate channeling. Our findings provide new guidance for enzyme orientation on 2 D scaffolds, which may be extrapolated to other multienzyme cascade systems.

Tunnelling current recognition through core-satellite gold nanoparticles for ultrasensitive detection of copper ions.

We report a new method for ultrasensitive detection of Cu(2+), which is based on changes in the tunnelling recognition current across self-assembled core-satellite gold nanoparticles (GNPs) networks functionalised with amino acids (l-cysteine). The addition of copper ions induces the formation of GNP/l-cysteine/Cu(2+)/l-cysteine/GNP molecular junctions and generates a significant decrease in the resistance through the networks. The networks are ultrasensitive to over ten orders range of copper ion concentrations.