RESUMO
The folding of proteins into intricate three-dimensional structures to achieve biological functions, such as catalysis, is governed by both kinetic and thermodynamic controls. The quest to design artificial enzymes using minimalist peptides seeks to emulate supramolecular structures existing in a catalytically active state. Drawing inspiration from the nuanced process of protein folding, our study explores the enzyme-like activity of amphiphilic peptide nanosystems in both equilibrium and non-equilibrium states, featuring the formation of supramolecular nanofibrils and nanosheets. In contrast to thermodynamically stable nanosheets, the kinetically trapped nanofibrils exhibit dynamic characteristics (e.g., rapid molecular exchange and relatively weak intermolecular packing), resulting in a higher hydrolase-mimicking activity. We emphasize that a supramolecular microenvironment characterized by an optimal local polarity, microviscosity, and ß-sheet hydrogen bonding is conducive to both substrate binding and ester bond hydrolysis. Our work underscores the pivotal role of both thermodynamic and kinetic control in impacting biomimetic catalysis and sheds a light on the development of artificial enzymes.
Assuntos
Hidrolases , Peptídeos , Peptídeos/química , Proteínas , Dobramento de Proteína , TermodinâmicaRESUMO
Catalytic generation of nitric oxide (NO) from NO donors by nanomaterials has enabled prolonged NO delivery for various biomedical applications, but this approach requires laborious synthesis routes. In this study, a new class of materials, that is, polymeric amines including polyethyleneimine (PEI), poly-L-lysine, and poly(allylamine hydrochloride), is discovered to induce NO generation from S-nitrosothiols (RSNOs) at physiological conditions. Controlled NO generation can be readily achieved by tuning the concentration of the NO donors (RSNOs) and polymers, and the type and molecular weight of the polymers. Importantly, the mechanism of NO generation by these polymers is deciphered to be attributed to the nucleophilic reaction between primary amines on polymers and the SNO groups of RSNOs. The NO-releasing feature of the polymers can be integrated into a suite of materials, for example, simply by embedding PEI into poly(vinyl alcohol) (PVA) hydrogels. The functionality of the PVA/PEI hydrogels is demonstrated for Pseudomonas aeruginosa biofilm prevention with a ≈4 log reduction within 6 h. As NO has potential therapeutic implications in various diseases, the identification of polymeric amines to induce NO release will open new opportunities in NO-generating biomaterials for antibacterial, antiviral, anticancer, antithrombotic, and wound healing applications.
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Óxido Nítrico , S-Nitrosotióis , Aminas/farmacologia , Doadores de Óxido Nítrico/farmacologia , Polímeros/farmacologia , Hidrogéis , S-Nitrosotióis/farmacologiaRESUMO
Autoimmune diseases and in particular type 1 diabetes rely heavily on treatments that target the symptoms rather than prevent the underlying disease. One of the barriers to better therapeutic strategies is the inability to detect and efficiently target rare autoreactive T-cell populations that are major drivers of these conditions. Here, we develop a unique artificial antigen-presenting cell (aAPC) system from biocompatible polymer particles that allows specific encapsulation of bioactive ingredients. Using our aAPC, we demonstrate that we are able to detect rare autoreactive CD4 populations in human patients, and using mouse models, we demonstrate that our particles are able to induce desensitization in the autoreactive population. This system provides a promising tool that can be used in the prevention of autoimmunity before disease onset.
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Diabetes Mellitus Tipo 1 , Linfócitos T , Animais , Células Apresentadoras de Antígenos , Autoimunidade , Linfócitos T CD4-Positivos , Diabetes Mellitus Tipo 1/terapia , Humanos , CamundongosRESUMO
Ceria nanoparticles (NPs) are widely reported to scavenge nitric oxide (NO) radicals. This study reveals evidence that an opposite effect of ceria NPs exists, that is, to induce NO generation. Herein, S-nitrosoglutathione (GSNO), one of the most biologically abundant NO donors, is catalytically decomposed by ceria NPs to produce NO. Ceria NPs maintain a high NO release recovery rate and retain their crystalline structure for at least 4 weeks. Importantly, the mechanism of this newly discovered NO generation capability of ceria NPs from GSNO is deciphered to be attributed to the oxidation of Ce3+ to Ce4+ on their surface, which is supported by X-ray photoelectron spectroscopy and density functional theory analysis. The prospective therapeutic effect of NO-generating ceria NPs is evaluated by the suppression of cancer cells, displaying a significant reduction of 93% in cell viability. Overall, this report is, to the authors' knowledge, the first study to identify the capability of ceria NPs to induce NO generation from GSNO, which overturns the conventional concept of them acting solely as a NO-scavenging agent. This study will deepen our knowledge about the therapeutic effects of ceria NPs and open a new route toward the NO-generating systems for biomedical applications.
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Cério , Nanopartículas , Catálise , Cério/química , Nanopartículas/química , Óxido Nítrico , S-NitrosoglutationaRESUMO
The highly diverse biological roles of nitric oxide (NO) in both physiological and pathophysiological processes have prompted great interest in the use of NO as a therapeutic agent in various biomedical applications. NO can exert either protective or deleterious effects depending on its concentration and the location where it is delivered or generated. This double-edged attribute, together with the short half-life of NO in biological systems, poses a major challenge to the realization of the full therapeutic potential of this molecule. Controlled release strategies show an admirable degree of precision with regard to the spatiotemporal dosing of NO but are disadvantaged by the finite NO deliverable payload. In turn, enzyme-prodrug therapy techniques afford enhanced deliverable payload but are troubled by the inherent low stability of natural enzymes, as well as the requirement to control pharmacokinetics for the exogenous prodrugs. The past decade has seen the advent of a new paradigm in controlled delivery of NO, namely localized bioconversion of the endogenous prodrugs of NO, specifically by enzyme mimics. These early developments are presented, successes of this strategy are highlighted, and possible future work on this avenue of research is critically discussed.
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Materiais Biomiméticos , Enzimas , Óxido Nítrico , Pró-Fármacos , Materiais Biomiméticos/química , Catálise , Sistemas de Liberação de Medicamentos , Enzimas/química , Humanos , Óxido Nítrico/química , Óxido Nítrico/metabolismo , Pró-Fármacos/químicaRESUMO
Early detection of cancer is likely to be one of the most effective means of reducing the cancer mortality rate. Hence, simple and ultra-quick methods for noninvasive detection of early-stage tumors are highly sought-after. In this study, a nanobiosensing platform with a rapid response time of nearly 30 s is introduced for the detection of matrilysin-the salivary gland cancer biomarker-with a limit of detection as low as 30 nm. This sensing platform is based on matrilysin-digestible peptides that bridge gold nanoparticle (AuNPs) cores (≈30-50 nm) and carbon quantum dot (CDs) satellites (≈9 nm). A stepwise synthesis procedure is used for self-assembly of AuNP-peptide-CDs, ensuring their long-term stability. The AuNP-peptide-CDs produce ideal optical signals, with noticeable fluorescence quenching effects. Upon peptide cleavage by matrilysin, CDs leave the surface of AuNPs, resulting in ultra-fast detectable violet and visible fluorescent signals.
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Técnicas Biossensoriais , Metaloproteinase 7 da Matriz/análise , Nanopartículas Metálicas , Neoplasias , Pontos Quânticos , Biomarcadores Tumorais/análise , Carbono , Ouro , Humanos , Limite de Detecção , PeptídeosRESUMO
Nitric oxide (NO) is a potent biological molecule that contributes to a wide spectrum of physiological processes. However, the full potential of NO as a therapeutic agent is significantly complicated by its short half-life and limited diffusion distance in human tissues. Current strategies for NO delivery focus on encapsulation of NO donors into prefabricated scaffolds or an enzyme-prodrug therapy approach. The former is limited by the finite pool of NO donors available, while the latter is challenged by the inherent low stability of natural enzymes. Zinc oxide (ZnO) particles with innate glutathione peroxidase and glycosidase activities, a combination that allows to catalytically decompose both endogenous (S-nitrosoglutathione) and exogenous (ß-gal-NONOate) donors to generate NO at physiological conditions are reported. By tuning the concentration of ZnO particles and NO prodrugs, physiologically relevant NO levels are achieved. ZnO preserves its catalytic property for at least 6 months and the activity of ZnO in generating NO from prodrugs in human serum is demonstrated. The ZnO catalytic activity will be beneficial toward generating stable NO release for long-term biomedical applications.
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Tecnologia Biomédica , Óxido Nítrico , Pró-Fármacos , Óxido de Zinco , Tecnologia Biomédica/métodos , Catálise , Humanos , Óxido Nítrico/síntese química , Doadores de Óxido Nítrico , Pró-Fármacos/química , Soro/química , Soro/enzimologia , Óxido de Zinco/químicaRESUMO
Synthetic micromotors are evaluated extensively in a range of biomedical, microscale transport, and environmental applications. Fundamental insight into micromotors that exhibit locomotion due to triggered disintegration of their associated liposomes is provided. Directed self-propulsion is observed when the lipid vesicles are solubilized using Triton X-100 (TX) and bile at sufficiently high concentrations. Directional motion, initiated by a propagating TX or bile gradient, is found when using a sufficiently high concentration of solubilization agents. On the other hand, a low bile concentration results in short-term reverse directional motion. The experimental and theoretical considerations offer valid fundamental understanding to complement the list of explored locomotion mechanisms for micromotors.
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Lipossomos , Locomoção , Movimento (Física) , OctoxinolRESUMO
Existing date marking tools, such as use-by and sell-by dates, fail to inform decision-making throughout food distribution because they do not adapt to storage conditions such as temperature. Polymer-based sensors can be incorporated into food packaging to provide an indication of food quality in real time, which can greatly reduce waste. This work identifies free fatty acid (FFA) as a marker for the quality of plant-based milk and demonstrates the first detection tool for the freshness of almond milk using phospholipid-doped polydiacetylene (PDA) vesicles. The sensor discriminates between triglycerides and FFA by a visible color change and can therefore be used to track fat metabolism during food spoilage. The interaction between FFA and PDA is investigated by electron microscopy and dynamic-light-scattering studies. PDA vesicles are then fabricated in agarose and used to discriminate between fresh and spoiled almond milk. Upon exposure of the PDA/agarose sensor to spoiled almond milk, a visible blue-to-red color change is induced in the film, which is correlated with FFA concentration in the samples. Incorporation of this technology into food packaging can be used to indicate food quality in real time, surpassing the efficacy of current date marking tools to reduce food waste.
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Colorimetria , Eliminação de Resíduos , Leite , Polímero PoliacetilênicoRESUMO
Membrane fusion is a process of fundamental importance in biological systems that involves highly selective recognition mechanisms for the trafficking of molecular and ionic cargos. Mimicking natural membrane fusion mechanisms for the purpose of biosensor development holds great potential for amplified detection because relatively few highly discriminating targets lead to fusion and an accompanied engagement of a large payload of signal-generating molecules. In this work, sequence-specific DNA-mediated liposome fusion is used for the highly selective detection of microRNA. The detection of miR-29a, a known flu biomarker, is demonstrated down to 18â nm within 30â min with high specificity by using a standard laboratory microplate reader. Furthermore, one order of magnitude improvement in the limit of detection is demonstrated by using a novel imaging technique combined with an intensity fluctuation analysis, which is coined two-color fluorescence correlation microscopy.
Assuntos
DNA/química , Transferência Ressonante de Energia de Fluorescência/métodos , Lipossomos/química , MicroRNAs/análise , Sequência de Bases , Biomarcadores/análise , Técnicas Biossensoriais/métodos , Humanos , Fusão de MembranaRESUMO
Multicompartmentalized microreactors are considered as cell mimics with hierarchical structures inspired by mammalian cells. We report the successful assembly and encapsulation of purified nuclei from RAW 264.7 cells (pNuc) into alginate-based microreactors. We demonstrate the preserved function of nuclei within the microreactors for mRNA production. Further, we load catalase-loaded liposomes (Lcat) into the microreactors to break down hydrogen peroxide (H2O2) into oxygen and water. Assemblies containing both natural pNuc and synthetic Lcat show significantly higher mRNA production in the presence of H2O2 compared to microreactors without Lcat or no H2O2 present, suggesting a beneficial effect of the locally enzymatically produced oxygen for transcription. This novel type of microreactors, containing both natural and synthetic compartments, presents a substantial advancement from assemblies equipped with solely synthetic units and offers opportunities as hypoxia models or for cell-free protein synthesis.
Assuntos
Reatores Biológicos , Catalase/metabolismo , Núcleo Celular/metabolismo , Hidrogéis/química , Peróxido de Hidrogênio/metabolismo , Lipossomos/química , RNA Mensageiro/metabolismo , Animais , Sobrevivência Celular , Células Cultivadas , Células Hep G2 , Humanos , Macrófagos/citologia , Macrófagos/metabolismo , Camundongos , Oxigênio , Engenharia TecidualRESUMO
We report the preparation of polymer capsules containing liposomal subcompartments, termed capsosomes, and their ability for the sustained delivery of protein therapeutics. Capsosomes were formed through the layer-by-layer (LbL) assembly of polymers and protein-loaded liposomes, followed by the formation of a capsule membrane based on disulfide cross-linked poly(methacrylic acid). The loading capacities of a model cargo (lysozyme) and brain-derived neurotrophic factor (BDNF), an important neurotrophin that has significant physiological functions on the nervous system, were determined, and the long-term release kinetics of the proteins was investigated in simulated physiological conditions. The capsosomes exhibited protein loading and release behavior that can be tuned by the lipid composition of the liposomal compartments, where inclusion of anionic lipids resulted in enhanced protein loading and slower release over the course of 80 days. These findings highlight the potential of capsosomes for the long-term delivery of protein therapeutics.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/química , Lipossomos/química , Muramidase/química , Fator Neurotrófico Derivado do Encéfalo/uso terapêutico , Cápsulas , Dissulfetos/química , Liberação Controlada de Fármacos , Glicerofosfolipídeos/química , Muramidase/uso terapêutico , Ácidos Polimetacrílicos/químicaRESUMO
Run Zhang, Songjun Zeng, and Rona Chandrawati introduce the Nanoscale Advances themed issue 'Nanomaterials for gas sensing and delivery'.
RESUMO
Colorimetric sensors play a crucial role in promoting on-site testing, enabling the detection and/or quantification of various analytes based on changes in color. These sensors offer several advantages, such as simplicity, cost-effectiveness, and visual readouts, making them suitable for a wide range of applications, including food safety and monitoring. A critical component in portable colorimetric sensors involves their integration with color models for effective analysis and interpretation of output signals. The most commonly used models include CIELAB (Commission Internationale de l'Eclairage), RGB (Red, Green, Blue), and HSV (Hue, Saturation, Value). This review outlines the use of color models via digitalization in sensing applications within the food safety and monitoring field. Additionally, challenges, future directions, and considerations are discussed, highlighting a significant gap in integrating a comparative analysis toward determining the color model that results in the highest sensor performance. The aim of this review is to underline the potential of this integration in mitigating the global impact of food spoilage and contamination on health and the economy, proposing a multidisciplinary approach to harness the full capabilities of colorimetric sensors in ensuring food safety.
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CRISPR-based diagnostics use the CRISPR-Cas system trans-cleavage activity to identify specific target sequences. When activated, this activity cleaves surrounding reporter molecules, producing a detectable signal. This technique has great specificity, sensitivity, and rapid detection, making it an important molecular diagnostic tool for medical and infectious disease applications. Despite its potential, the present CRISPR/Cas system has challenges with its single-stranded DNA reporters, characterized by low stability and limited sensitivity, restricting effective application in complex biological settings. In this work, we investigate the trans-cleavage activity of CRISPR/Cas12a on substrates utilizing fluorescent polystyrene microspheres to detect tetracycline. This innovative discovery led to the development of microsphere probes addressing the stability and sensitivity issues associated with CRISPR/Cas biosensing. By attaching the ssDNA reporter to polystyrene microspheres, we discovered that the Cas12a system exhibits robust and sensitive trans-cleavage activity. Further work revealed that the trans-cleavage activity of Cas12a on the microsphere surface is significantly dependent on the concentration of the ssDNA reporters. Building on these intriguing discoveries, we developed microsphere-based fluorescent probes for CRISPR/Cas aptasensors, which showed stability and sensitivity in tetracycline biosensing. We demonstrated a highly sensitive detection of tetracycline with a detection limit of 0.1 µM. Finally, the practical use of a microsphere-based CRISPR/Cas aptasensor in spiked food samples was proven successful. These findings highlighted the remarkable potential of microsphere-based CRISPR/Cas aptasensors for biological research and medical diagnosis.
Assuntos
Técnicas Biossensoriais , Sistemas CRISPR-Cas , DNA de Cadeia Simples , Microesferas , Poliestirenos , Tetraciclina , Tetraciclina/análise , Sistemas CRISPR-Cas/genética , Poliestirenos/química , DNA de Cadeia Simples/química , DNA de Cadeia Simples/genética , Técnicas Biossensoriais/métodos , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/metabolismo , Proteínas Associadas a CRISPR/genética , Proteínas Associadas a CRISPR/metabolismo , Corantes Fluorescentes/química , Proteínas de Bactérias , EndodesoxirribonucleasesRESUMO
Neurodegenerative disorders pose a significant challenge to global healthcare, with Alzheimer's disease (AD) being one of the most prevalent forms. Early and accurate detection of amyloid-ß (Aß) (1-42) monomers, a key biomarker of AD pathology, is crucial for effective diagnosis and intervention of the disease. Current gold standard detection techniques for Aß include enzyme-linked immunosorbent assay and surface plasmon resonance. Although reliable, they are limited by their cost and time-consuming nature, thus restricting their point-of-care applicability. Here we present a sensitive and rapid colorimetric sensor for the detection of Aß (1-42) monomers within 5 min. This was achieved by harnessing the peroxidase-like activity of metal-loaded metal-organic frameworks (MOFs), specifically UiO-66-NH2, coupled with the strong affinity of Aß (1-42) to the MOFs. Various metal-loaded MOFs were synthesized and investigated, and platinum-loaded UiO-66-NH2 was identified as the optimal candidate for our purpose. The Pt-loaded UiO-66-NH2 sensor demonstrated detection limits of 2.76 and 4.65 nM Aß (1-42) monomers in water and cerebrospinal fluid, respectively, with a linear range from 0.75 to 25 nM (R2 = 0.9712), outperforming traditional detection techniques in terms of both detection time and complexity. Moreover, the assay was specific toward Aß (1-42) monomers when evaluated against interfering compounds. The rapid and cost-effective sensor may help circumvent the limitations of conventional detection methods, thus providing a promising avenue for early AD diagnosis and facilitating improved clinical outcomes.
Assuntos
Doença de Alzheimer , Estruturas Metalorgânicas , Compostos Organometálicos , Ácidos Ftálicos , Humanos , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/líquido cefalorraquidiano , Peptídeos beta-Amiloides/líquido cefalorraquidiano , BiomarcadoresRESUMO
Nitric oxide (NO) is a key signalling molecule released by vascular endothelial cells that is essential for vascular health. Low NO bioactivity is associated with cardiovascular diseases, such as hypertension, atherosclerosis, and heart failure and NO donors are a mainstay of drug treatment. However, many NO donors are associated with the development of tolerance and adverse effects, so new formulations for controlled and targeted release of NO would be advantageous. Herein, we describe the design and characterisation of a novel NO delivery system via the reaction of acidified sodium nitrite with thiol groups that had been introduced by cysteamine conjugation to porous graphene oxide nanosheets, thereby generating S-nitrosated nanosheets. An NO electrode, ozone-based chemiluminescence and electron paramagnetic resonance spectroscopy were used to measure NO released from various graphene formulations, which was sustained at >5 × 10-10 mol cm-2 min-1 for at least 3 h, compared with healthy endothelium (cf. 0.5-4 × 10-10 mol cm-2 min-1). Single cell Raman micro-spectroscopy showed that vascular endothelial and smooth muscle cells (SMCs) took up graphene nanostructures, with intracellular NO release detected via a fluorescent NO-specific probe. Functionalised graphene had a dose-dependent effect to promote proliferation in endothelial cells and to inhibit growth in SMCs, which was associated with cGMP release indicating intracellular activation of canonical NO signalling. Chemiluminescence detected negligible production of toxic N-nitrosamines. Our findings demonstrate the utility of porous graphene oxide as a NO delivery vehicle to release physiologically relevant amounts of NO in vitro, thereby highlighting the potential of these formulations as a strategy for the treatment of cardiovascular diseases.
Assuntos
Grafite , Óxido Nítrico , Grafite/química , Óxido Nítrico/metabolismo , Humanos , Nanoestruturas/química , Porosidade , Doadores de Óxido Nítrico/química , Doadores de Óxido Nítrico/farmacologia , Doadores de Óxido Nítrico/administração & dosagem , Proliferação de Células/efeitos dos fármacos , Doenças Cardiovasculares/tratamento farmacológico , Células Endoteliais/metabolismo , Células Endoteliais/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacosRESUMO
We report that the continuous assembly of polymers (CAP) approach, mediated by ring-opening metathesis polymerization (ROMP), is a facile and versatile technology to prepare engineered nanocoatings for various biomedical applications. Low-fouling coatings on particles were obtained by the formation of multicompositional, layered films via simple and efficient tandem CAP(ROMP) processes that are analogous to chain extension reactions. In addition, the CAP(ROMP) approach allows for the efficient postfunctionalization of the CAP films with bioactive moieties via cross-metathesis reactions between the surface-immobilized catalysts and symmetrical alkene derivatives. The combined features of the CAP(ROMP) approach (i.e., versatile polymer selection and facile functionalization) allow for the fabrication and surface modification of various types of polymer films, including those with intrinsic protein-repellent properties and selective protein recognition capabilities. This study highlights the various types of advanced coatings and materials that the CAP approach can be used to generate, which may be useful for biomedical applications.
Assuntos
Resinas Acrílicas/química , Materiais Revestidos Biocompatíveis/química , Acrilatos/química , Adsorção , Avidina/química , Biotina/química , Reagentes de Ligações Cruzadas , Polietilenoimina/química , Porosidade , Ligação Proteica , Soroalbumina Bovina/química , Propriedades de SuperfícieRESUMO
The detection of pathogenic bacteria is essential to prevent and treat infections and to provide food security. Current gold-standard detection techniques, such as culture-based assays and polymerase chain reaction, are time-consuming and require centralized laboratories. Therefore, efforts have focused on developing point-of-care devices that are fast, cheap, portable and do not require specialized training. Paper-based analytical devices meet these criteria and are particularly suitable to deployment in low-resource settings. In this Review, we highlight paper-based analytical devices with substantial point-of-care applicability for bacteria detection and discuss challenges and opportunities for future development.
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Liposomes and polymersomes have attracted significant attention and have emerged as versatile materials for therapeutic delivery and in the design of artificial cells and organelles. Through the judicious choice of building blocks, these synthetic carriers can be readily engineered with tailored interfacial properties, offering new possibilities for the design of advanced assemblies with specific permeability, stability, stimuli response, and targeting capabilities. In this feature article, we highlight recent studies on biomimetic liposome- and polymersome-based multicompartmentalized assemblies en route toward the development of artificial cells, microreactors, and therapeutic delivery carriers. The strategies employed to produce these carriers are outlined, and the properties that contribute to their performance are discussed. Applications of these biomimetic assemblies are highlighted, and finally, areas that require additional investigation for the future development of these assemblies as next-generation therapeutic systems are outlined.