Applications In Biological Environments Research Articles

A near-infrared emitting “off-on” fluorescent probe for bioimaging of Pd(Ⅱ) ions in living cells and mice

BackgroundThe surging consumption of palladium in modern industry has given rise to its accumulation in the ecosystem, posing conspicuous toxicity to aquatic organisms and human health. The investigation of palladium in biological systems is highly demanded for the in-depth understanding of its dynamics and behaviors. Fluorescence imaging serves as a powerful approach to assess palladium species in biological systems, and currently most of the sensing probes are applicable to living cells. Effective tracking of palladium species in living organisms is challenging, which requires sufficient hydrophilicity and imaging depth of the probes. ResultsBased on an intramolecular charge transfer (ICT) mechanism, a distyryl boron dipyrromethene (BODIPY) derivative (DISBDP-Pd) has been prepared for the near-infrared (NIR) fluorescence imaging of Pd2+ ions. Two additional methoxy triethylene glycol (TEG) chains could serve as flexible and hydrophilic moieties to enhance the aqueous solubility and cell permeability of the extended conjugate. Solution studies revealed that DISBDP-Pd exhibited a NIR fluorescence enhancement signal exclusively to Pd2+ ions (detection limit as low as 0.85 ppb) with negligible interference from Pd0 species and other closely related metal ions. Computational calculations have been performed to rationalize the binding mode and the mechanism of action. Fluorescence imaging assays have been conducted on A549 human non-small cell lung carcinoma cells and mouse models. Exhibiting negligible cytotoxicity, DISBDP-Pd demonstrated concentration-related fluorescence enhancement signals in response to Pd2+ ions in living cells and mice. SignificanceDISBDP-Pd exhibits advantages over many small molecule palladium probes in terms of satisfactory aqueous solubility, high sensitivity and selectivity, and biocompatible NIR emission property, which are particularly favorable for the sensing application in biological environments. The design strategy of this probe can potentially be adopted for the functionalization of other BODIPY probes implemented for NIR fluorescence bioimaging.

Synthesis of a novel photochemical and thermoresponsive diblock biomaterial with end-functionalized zinc porphyrin.

Porphyrin compound-based photochemical molecules and biomaterials have been synthesized for photosensitivity and bioimaging experiments. However, most porphyrin photosensitizers have limited application in biological environments owing to severe aggregation in aqueous solutions. In the present study, we prepared amphipathic and photosensitive copolymers using zinc porphyrin via consecutive atom transfer-free radical polymerizations (ATRPs) comprising photoresponsive and thermosensitive chain segments. Furthermore, we evaluated the photocatalytic activity of the copolymer for methylene blue (MB) in water. Methods: First, we synthesized a photoresponsive ain segment of poly (6-[4-(4-methoxyphenylazo)phenoxy]hexyl methacrylate) (ZnPor-PAzo); then, ZnPor-PAzo was used as a macroinitiator and was polymerized with N-isopropylacrylamide (NIPAM) via ATRPs to obtain a novel photochemical and thermoresponsive diblock biomaterial with end-functionalized zinc porphyrin [(ZnPor-PAzo)-PNIPAMs]. Results: The polydispersity index (M w/M n) of (ZnPor-PAzo)-PNIPAMs was 1.19-1.32. Furthermore, its photoresponsive and thermosensitive characteristics were comprehensively studied. Discussion: The end-functionalized diblock copolymer (ZnPor-PAzo)-PNIPAM exhibits obvious fluorescence and efficient photocatalytic activity for aqueous MB under visible light.

Review of: "nanobiosensors (biological nanosensors) due to their nanometer size and application in biological environments"

Review of: "nanobiosensors (biological nanosensors) due to their nanometer size and application in biological environments"

Biomembrane‐Based Memcapacitive Reservoir Computing System for Energy‐Efficient Temporal Data Processing

Reservoir computing is a highly efficient machine learning framework for processing temporal data by extracting input features and mapping them into higher dimensional spaces. Physical reservoirs have been realized using spintronic oscillators, atomic switch networks, volatile memristors, etc. However, these devices are intrinsically energy‐dissipative due to their resistive nature, increasing their power consumption. Therefore, memcapacitive devices can provide a more energy‐efficient approach. Herein, volatile biomembrane‐based memcapacitors are leveraged as reservoirs to solve classification tasks and process time series in simulation and experimentally. This system achieves a 99.6% accuracy for spoken‐digit classification and a normalized mean square error of in a second‐order nonlinear regression task. Furthermore, to showcase the device's real‐time temporal data processing capability, a 100% accuracy for an epilepsy detection problem is achieved. Most importantly, it is demonstrated that each memcapacitor consumes an average of 41.5 fJ of energy per spike, regardless of the selected input voltage pulse width, while maintaining an average power of 415 fW for a pulse width of 100 ms, orders of magnitude lower than those achieved by state‐of‐the‐art devices. Lastly, it is believed that the biocompatible, soft nature of our memcapacitor renders it highly suitable for computing applications in biological environments.

Designing Covalent Organic Framework-based Light-driven Microswimmers towards Therapeutic Applications.

While micromachines with tailored functionalities enable therapeutic applications in biological environments, their controlled motion and targeted drug delivery in biological media require sophisticated designs for practical applications. Covalent organic frameworks (COFs), a new generation of crystalline and nanoporous polymers, offer new perspectives for light-driven microswimmers in heterogeneous biological environments including intraocular fluids, thus setting the stage for biomedical applications such as retinal drug delivery. We describe and compare two different types of COFs, uniformly spherical TABP-PDA-COF sub-micron particles and texturally nanoporous, micron-sized TpAzo-COF particles as light-driven microrobots. They can be used as highly efficient visible-light-driven drug carriers in aqueous ionic and cellular media. Their absorption ranging down to red light enables phototaxis even in deeper and viscous biological media, while the organic nature of COFs ensures their biocompatibility. Their inherently porous structures with ∼2.6 and ∼3.4nm pores, and large surface areas allow for targeted and efficient drug loading even for insoluble drugs, which can be released on demand. Additionally, indocyanine green (ICG) dye loading in the pores enables photoacoustic imaging, optical coherence tomography, and hyperthermia in operando conditions. This real-time visualization of the drug-loaded COF microswimmers enables unique insights into the action of photoactive porous drug carriers for therapeutic applications. This article is protected by copyright. All rights reserved.

Nonbiodegradable Spiegelmer-Driven Colorimetric Biosensor for Bisphenol A Detection.

Spiegelmers are enantiomers of natural D-oligonucleotides that bind to targets with distinct structures such as aptamers. The high susceptibility of natural D-form aptamers to nucleases greatly hinders their application in biological environments. Here, a nonbiodegradable spiegelmer-based platform for the sensitive detection of bisphenol A (BPA) was developed. Due to the symmetric molecule of BPA, the D-form aptamer can be directly converted into mirror forms via chemical synthesis. Aptamer–target interactions that involve chemically synthesized spiegelmers were characterized by biolayer interferometry, and their stabilities were tested in various biological fluids by exposure to nucleases. We demonstrate for the first time the use of a nuclease-resistant spiegelmer in a simple, label-free gold nanoparticle-based colorimetric assay to detect BPA in a highly sensitive and selective manner. The aptasensor exhibits an LOD of 0.057 ng/mL and dynamic range of 105 (100 pg/mL to 10 mg/mL). With sensing capacity and biological stability, the developed aptasensor shows great potential to utilize in in-field applications such as water quality monitoring.

A Learning Automaton-Based Algorithm for Maximizing the Transfer Data Rate in a Biological Nanonetwork

Biological nanonetworks have been envisaged to be the most appropriate alternatives to classical electromagnetic nanonetworks for applications in biological environments. Due to the diffusional method of the message exchange process, transfer data rates are not proportional to their electromagnetic counterparts. In addition, the molecular channel has memory affecting the reception of a message, as the molecules from previously transmitted messages remain in the channel, affecting the number of information molecules that are required from a node to perceive a transmitted message. As a result, the ability of a node to receive a message is directly connected to the transmission rate from the transmitter. In this work, a learning automaton approach has been followed as a way to provide the receiver nodes with an algorithm that could firstly enhance their reception capability and secondly boost the performance of the transfer data rate between the biological communication parties. To this end, a complete set of simulation scenarios has been devised, simulating different distances between nodes and various input signal distributions. Most of the operational parameters, such as the speed of convergence for different numbers of ascension and descension steps and the number of information molecules per message, have been tested pertaining to the performance characteristics of the biological nanonetwork. The applied analysis revealed that the proposed protocol manages to adapt to the communication channel changes, such as the number of remaining information molecules, and can be successfully employed at nanoscale dimensions as a tool for pursuing an increased transfer data rate, even with time-variant channel characteristics.

Tuning the photoreactivity of photocycloaddition by halochromism

Harnessing the power of light for chemical transformation is a long-standing goal in organic synthesis, materials fabrication and engineering. Amongst all photochemical reactions, [2 + 2] photocycloadditions are inarguably the most important and most frequently used. These photoreactions have green characteristics by enabling new bond formation in a single step procedure under light irradiation, without the need for heat or chemical catalysis. More recently, substantial progress has been made in red-shifting the activation wavelength of photocycloadditions in response to research trends moving towards green and sustainable processes, and advanced applications in biological environments. In the past 5 years, our team has further expanded the toolbox of photocycloaddition reactions that can be triggered by visible light. In our exploration of photochemical reactivity, we found that reactivity is often red-shifted compared to the substrate’s absorption spectrum. Our efforts have resulted in red-shifted photochemical reactions, providing some of the lowest energy – and catalyst-free – photo-activated [2 + 2] cycloadditions (up to 550 nm). More recently, we introduced an additional level of control over such finely wavelength gated reactions by altering the pH of the reaction environment, thus exploiting halochromic effects to enhance or impede the photoreactivity of red-shifted [2 + 2] photocycloaddition reactions. In this account, we discuss the current state of halochromically regulated photochemical reactions and their potential in soft matter materials on selected examples.

Graphene oxide induced enhancement of light-driven micromotor with biocompatible fuels

A highly efficient light-driven cuprous oxide @ graphene oxide (Cu2O@GO) micromotor which can be propelled with various biocompatible fuels is described. The Cu2O@GO micromotors, which achieve enhanced photocatalytic performance by doping Cu2O with GO, can be efficiently propelled by visible light and even near infrared light. Compared with conventional plain Cu2O micromotors, which can only be driven by visible light, the Cu2O@GO micromotors show more than 3 times faster speeds under the full range of visible light with biocompatible glucose fuel. In addition, due to the GO-induced enhanced photocatalytic performance and local photoinduced thermal effects, such motors can be powered by NIR with a series of biocompatible fuels, such as glucose, leucine, and urea solutions, with speeds reaching up to 11.39 µm s−1 (in glucose), which extends the excitation wavelength of photocatalytic micromotors to near-infrared light. This novel strategy of improving the performance of light-driven micromotors via GO doping has opened up a new and attractive path for the preparation of micromotors with potential applications in biological environments.

A Perspective on Application of Carbon Quantum Dots in Luminescence Immunoassays.

Quantum dots (QDs) have been established in our daily life, for instance, in display screens and light-harvesting technologies, mainly owing to their peculiar opto-electronic features. However, toxicity of inorganic QDs, such as CdSe, CdTe, and perovskites, limits their applications in biological environments for medical diagnosis and bio-imaging purposes. A new generation of QDs called carbon quantum dots (CQDs) have been progressing rapidly over the past few years. CQDs have become as popular as other carbon-based nanomaterials such as carbon nanotubes (CNTs), due to their ease of preparation, ultra-small size, biocompatibility, and bright luminescence.

Cooperation of ESIPT and ICT Processes in the Designed 2-(2'-Hydroxyphenyl)benzothiazole Derivative: A Near-Infrared Two-Photon Fluorescent Probe with a Large Stokes Shift for the Detection of Cysteine and Its Application in Biological Environments.

A rationally designed near-infrared two-photon fluorescent probe (SDP-A) for selectively detecting cysteine (Cys) has been developed based on a newly designed conjugation-enhanced 2-(2'-hydroxyphenyl)benzothiazole derivative as the fluorophore, an acrylate moiety as the Cys reaction site, and an N-methylpyridinium scaffold as both the unit of organelle targeting and improving water solubility. The probe SDP-A alone essentially emitted no fluorescence, whereas it achieved a superb near-infrared fluorescence emission (713 nm) enhancement within 15 min with a significant Stokes shift (302 nm) in the presence of Cys. The photoluminescence mechanism of the probe SDP-A toward Cys was modulated by excited-state intramolecular proton transfer (ESIPT) and intramolecular charge transfer (ICT) processes. It exhibited high selectivity and sensitivity (LOD = 102 nM) for monitoring Cys over other analytes such as Hcy/GSH/H2S owing to a specific conjugate addition-cyclization reaction between Cys and the acrylate moiety. More importantly, the released fluorophore SDP exhibits elevated quantum yields (1.52-18.17%) in different solvents and strong two-photon excited fluorescence with a sizeable two-photon action cross-section (Φ) of 213.5 GM at 820 nm in acetonitrile-PBS medium, which is highly desirable for two-photon fluorescence imaging of the living samples. Therefore, SDP-A was successfully applied to the imaging of Cys in live cells, zebrafish, mouse brain, and abdominal cavity down to a depth of more than 200 μm using a one/two-photon fluorescence microscope.

Catalytic DNA Strand Displacement Cascades Applied to Logic Programming

The field of DNA computing is devoted to the creation of devices capable of processing information signals encoded on biological substrates. These signals are intended to propagate in cascades of biochemical reactions in which they naturally undergo a progressive reduction. Preventing signal reduction becomes crucial considering applications in biological environments where molecular cues are scarce. Although catalytic gates have been developed using the toehold-exchange mechanism for logic gate circuits, the matter remains unaddressed for logic reasoning devices. Inspired by the main work in biomolecular logic programming, we present a new encoding scheme for facts, rules, and queries to implement backward/forward chaining inference paths via catalytic DNA strand displacement cascades. In this context, we take advantage of fueling reactions to recover inputs, which preserve their availability to react with different implication gates. Our molecular design is thermodynamically analyzed by providing suitable sequences for the correct formation of structures. With regard to the kinetic performance, data from simulations suggest that the model operates efficiently even with identified crosstalk reactions.

Encapsulation of AgNPs within Zwitterionic Hydrogels for Highly Efficient and Antifouling Catalysis in Biological Environments.

Silver nanoparticles (AgNPs) have been widely used as catalysts in a variety of chemical reactions owing to their unique surface and electronic properties, but their practical applications have been hindered by severe aggregation. The immobilization of AgNPs is crucial to preventing their aggregation or precipitation as well as to improving their reusability. Herein, we developed a facile route for the reductant-free in situ synthesis of AgNPs in zwitterionic hydrogels. Via this method, the embedded AgNPs had a uniform distribution, high activity, and antibiofouling capability. The catalytic reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP) using polycarboxybetaine-AgNPs (PCB-AgNPs) could achieve >95% conversion efficiency within 5 min. Meanwhile, the normalized rate constant knor (10.617 s-1mmol-1) was higher than that of most of the reported immobilized nanocatalysts. More importantly, in a biofouling environment, PCB-AgNPs could still exhibit >97% initial catalytic activity while AgNPs in the PSB or PHEMA hydrogel lost ∼60% activity. This strategy holds great potential for the immobilization of nanoparticle catalysts, especially for applications in biological environments.

Preparation of carbon dots with long-wavelength and photoluminescence‐tunable emission to achieve multicolor imaging in cells

Most carbon dots (CDs) derived from natural biowastes show emission limitedly in the blue range. This work applied urea and sulfuric acid as dopants to assist the carbonization of cellulose-based biowaste willow catkin. Then a facile chemical cutting method was developed for scalable production of CDs. Under excitation at a single-wavelength of 360 nm, the aqueous solutions of CDs obtained at different cutting temperatures (100, 120, and 140 °C) exhibited strong blue (486 nm), green (522 nm) and yellow (545 nm) emission, respectively. The as-prepared CDs also exhibited excitation-independent emission behaviors and high photostability in different pH and ionic strength. Furthermore, these long-wavelength and photoluminescence-tunable CDs were successfully applied to multicolor bioimaging in Hela cells, indicating their potential toward multimodal sensing applications in biological environment.

Electrodeposition of Zwitterionic PEDOT Films for Conducting and Antifouling Surfaces.

Conferring antifouling properties can extend the use of conducting polymers in biosensors and bioelectronics under complex biological conditions. On the basis of the antifouling properties of a series of zwitterionic polymers, we synthesized new thiophene-based compounds bearing a phosphorylcholine, carboxybetaine, or sulfobetaine pendant group. The monomers were synthesized by a facile reaction of thiol-functionalized 3,4-ethylenedioxythiophene with zwitterionic methacrylates. Electrochemical copolymerization was performed to deposit zwitterionic poly(3,4-ethylenedioxythiophene) (PEDOT) films with tunable conducting and antifouling properties on a conducting substrate. Electrochemical impedance spectroscopy showed that the conductivity and capacitance decreased with increasing zwitterionic content in the films. Protein adsorption and cell adhesion studies showed the effects of the type and content of zwitterions on the antifouling characteristics. Optimization of the electrodeposition conditions enabled development of both conducting and antifouling polymer films. These antifouling conjugated functional polymers have promising applications in biological environments.

Advances in surface-coated single-walled carbon nanotubes as near-infrared photoluminescence emitters for single-particle tracking applications in biological environments

Single-particle tracking (SPT) represents a powerful tool for revealing the single-molecule dynamics in a number of biological processes in live cells and biological tissue. Single-walled carbon nanotubes (SWCNTs) are promising photoluminescence emitters for SPT applications in various biological environments due to their characteristic large-aspect-ratio structures along with their bright and stable near-infrared (NIR) photoluminescence, which are invaluable for long-term video-rate imaging and tracking applications at the single-molecule level with high signal-to-noise ratios (SNRs). Recent advances in applying SWCNTs as NIR photoluminescence emitters have highlighted the understanding of brain tissue organization at the nanometer scale. In the first section, this review article summarizes the latest advances in different surface coatings commonly used for encapsulating SWCNT surfaces via molecular self-assembly in order to obtain surface-coated nanotubes with low cytotoxicity and minimal nonspecific interactions with live cells while maintaining their emission of bright photoluminescence to enable long-term photoluminescent imaging and tracking at the single-nanotube level in biological environments. The second section offers a comparison of different excitation strategies of (6,5) SWCNTs to determine the best excitation wavelength for efficient video-rate imaging and tracking of individual nanotubes in live brain tissue for up to tens of minutes without inducing unacceptable phototoxicity or temperature increases. Finally, this review showcases that, by utilizing the photoluminescence tracking of single nanotubes combined with super-resolution single-molecule localization microscopy technologies, it is practical to elucidate the ultrafine nanometer-scale organization of the brain extracellular space (ECS) and probe the local rheological properties of young rat brain with a subdiffraction optical resolution down to 50 nm at a subwavelength accuracy of ~40 nm. The findings primarily indicate the great diversity of the brain ECS and the inhomogeneous properties of the local viscosity in live brain tissue. Overall, because of their advantages of low cytotoxicity, bright photoluminescence, high SNRs (~25), and deep tissue penetration (~100 μm) for long-term video-rate imaging and tracking at the single-nanotube level under 845 nm excitation (K-momentum exciton–phonon sideband, KSB), phospholipid-polyethylene glycol-coated SWCNTs hold great potential as NIR photoluminescence emitters for single-particle tracking in biological environments, as exemplified here in live brain tissue, and may find extended applications in elucidating the fundamental roles of the brain ECS in various biological processes, such as sleep, memory, aging, brain tumor progression, and neurodegenerative disease development.

Photoswitching of DNA Hybridization Using a Molecular Motor.

Reversible control over the functionality of biological systems via external triggers may be used in future medicine to reduce the need for invasive procedures. Additionally, externally regulated biomacromolecules are now considered as particularly attractive tools in nanoscience and the design of smart materials, due to their highly programmable nature and complex functionality. Incorporation of photoswitches into biomolecules, such as peptides, antibiotics, and nucleic acids, has generated exciting results in the past few years. Molecular motors offer the potential for new and more precise methods of photoregulation, due to their multistate switching cycle, unidirectionality of rotation, and helicity inversion during the rotational steps. Aided by computational studies, we designed and synthesized a photoswitchable DNA hairpin, in which a molecular motor serves as the bridgehead unit. After it was determined that motor function was not affected by the rigid arms of the linker, solid-phase synthesis was employed to incorporate the motor into an 8-base-pair self-complementary DNA strand. With the photoswitchable bridgehead in place, hairpin formation was unimpaired, while the motor part of this advanced biohybrid system retains excellent photochemical properties. Rotation of the motor generates large changes in structure, and as a consequence the duplex stability of the oligonucleotide could be regulated by UV light irradiation. Additionally, Molecular Dynamics computations were employed to rationalize the observed behavior of the motor–DNA hybrid. The results presented herein establish molecular motors as powerful multistate switches for application in biological environments.

Fused electron deficient semiconducting polymers for air stable electron transport

Conventional semiconducting polymer synthesis typically involves transition metal-mediated coupling reactions that link aromatic units with single bonds along the backbone. Rotation around these bonds contributes to conformational and energetic disorder and therefore potentially limits charge delocalisation, whereas the use of transition metals presents difficulties for sustainability and application in biological environments. Here we show that a simple aldol condensation reaction can prepare polymers where double bonds lock-in a rigid backbone conformation, thus eliminating free rotation along the conjugated backbone. This polymerisation route requires neither organometallic monomers nor transition metal catalysts and offers a reliable design strategy to facilitate delocalisation of frontier molecular orbitals, elimination of energetic disorder arising from rotational torsion and allowing closer interchain electronic coupling. These characteristics are desirable for high charge carrier mobilities. Our polymers with a high electron affinity display long wavelength NIR absorption with air stable electron transport in solution processed organic thin film transistors.

Evaluation of Different Single-Walled Carbon Nanotube Surface Coatings for Single-Particle Tracking Applications in Biological Environments.

Fluorescence imaging of biological systems down to the single-molecule level has generated many advances in cellular biology. For applications within intact tissue, single-walled carbon nanotubes (SWCNTs) are emerging as distinctive single-molecule nanoprobes, due to their near-infrared photoluminescence properties. For this, SWCNT surfaces must be coated using adequate molecular moieties. Yet, the choice of the suspension agent is critical since it influences both the chemical and emission properties of the SWCNTs within their environment. Here, we compare the most commonly used surface coatings for encapsulating photoluminescent SWCNTs in the context of bio-imaging applications. To be applied as single-molecule nanoprobes, encapsulated nanotubes should display low cytotoxicity, and minimal unspecific interactions with cells while still being highly luminescent so as to be imaged and tracked down to the single nanotube level for long periods of time. We tested the cell proliferation and cellular viability of each surface coating and evaluated the impact of the biocompatible surface coatings on nanotube photoluminescence brightness. Our study establishes that phospholipid-polyethylene glycol-coated carbon nanotube is the best current choice for single nanotube tracking experiments in live biological samples.

Cu-bearing, martensitic stainless steels for applications in biological environments

Precipitation of Cu in stainless steels disturbs the passivating oxide layer and intensifies the release of Cu ions in biological environments which act toxically against the microorganisms. The phenomenon allows promisingly the production of age hardenable, Cu-bearing stainless steels with antibacterial property, suitable for fabrication of medical, home and environmental appliances. In the present study, aging behavior, microstructure and antibacterial performance of AISI 410s stainless steel with different amounts of Cu addition (nominally 1, 3 and 5wt%) were investigated. Microhardness, X-ray diffraction, optical, scanning and transmission electron microscopy were used. After solution annealing at 1330K and oil quenching, the AISI 410s steel showed dual phase martensite-ferrite microstructure. It was found that increasing of Cu content decreases ferrite content remarkably. Both of martensite and ferrite phase constituents showed remarkable precipitation hardening after aging at 773K. Their age hardenability increases with increasing of Cu content. Antibacterial performance was evaluated against Escherichia coli and Staphylococcus aureus bacteria, the common pathogens of human disease and food poisoning. Addition of 3wt% Cu gives appropriate age hardenability and antibacterial performance.