Recent progress in molecular diagnostics has been strongly influenced by advances in magnetic bead (MB) chemistry. Synthetic strategies for MB fabrication play a critical role in defining their size, physicochemical properties, and nano‐ or microscale architectures, which ultimately determine their analytical performance. The efficiency of MBs in liquid biopsy depends on the architecture of their components, including magnetic core, surface functionalization, as well as the integration of recognition ligands. This review highlights the recent developments in MB design for the selective capture, identification, and quantification of cancer biomarkers in liquid biopsy applications. It also summarizes the advantages and limitations of commercially available MB platforms and critically evaluates emerging systems reported in the literature. By comparing current technologies, this review discusses major advances as well as remaining translational bottlenecks, providing guidance for the rational design of next‐generation MBs for liquid biopsy. The latter is an emerging technology increasingly used in precision oncology for molecular profiling to support cancer diagnosis, prognosis, and the selection of personalized treatments.

In the original publication [...].

Graphene-based nanomaterials exhibit exceptional physicochemical properties that facilitate a range of diverse biomedical applications, including liquid biopsy. In this study, graphene-based magnetic units, termed MAGU (MAGnetic Units), were specifically engineered for the selective isolation of exosomes. Total extracellular vesicles were first enriched using ultracentrifugation, followed by immunomagnetic capture of CD9+ exosomes. MAGU functionalized with anti-CD9 antibody (MAGU-anti-CD9) efficiently recovered a CD9-positive exosome subpopulation expressing canonical markers ALIX, CD147, TSG101, and Flotillin-1, thereby confirming selective isolation performance. To investigate viral associated signaling, 293T cells were transduced with SARS-CoV-2 spike pseudovirus. This pseudovirus was engineered to express the SARS-CoV-2 spike protein, enabling simulation of viral entry and assessment of potential alterations in the exosomal profile induced by viral binding. Exosomes released by pseudovirus-transduced 293T cells were analyzed and compared to those from non-transduced controls. The MAGU-anti-CD9 complex selectively isolated a defined subset of CD9-positive vesicles enriched in the multifunctional transmembrane glycoprotein CD147, which has been proposed as a cofactor in SARS-CoV-2 entry. Comprehensive molecular profiling of selectively captured exosome subpopulations is expected to further support the application of MAGU technology in virus-host interaction research and liquid-biopsy-based diagnostics.

Polymeric nanoparticles bearing multifunctional features, including surface functionalization, hold great promise to address current limitations in antimicrobial drug delivery. Among bioactive ligands, N-acetylcysteine (NAC) and its derivatives have been investigated for their ability to interfere with biofilm formation and to disrupt mature biofilms. Here we propose novel N-acetylcysteine-decorated Nanoantibiotics based on poly(lactic acid)poly(ethylene glycol) nanoparticles (PLA-PEG-NAC NPs) incorporating Linezolid (LNZ) and Pentamidine (Pent). NAC was selected to target methicillin-resistant Staphylococcus aureus (MRSA) biofilm and Pent for broadening the antimicrobial spectrum of LNZ. The PLA-PEG-NAC copolymer was synthesized through a multi-step pathway involving carbonyldiimidazole-mediated amidation, Steglich esterification, and copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC). Successful NAC grafting was confirmed by thiol-ene Michael addition using a crotonylsubstituted BODIPY dye. Nanoparticle's formulation by dialysis achieved encapsulation efficiencies of 12% for LNZ and 20% for Pent, with sustained drug release profiles (78% and 25% cumulative release within 24 h, respectively). Despite the moderate encapsulation efficiency, the observed biological effects closely reflected the drug release profile and the achieved drug loading proved suitable for the aims of the present work. Cytotoxicity assays in Vero cells demonstrated no notable toxicity of PLA-PEG-NAC NPs, either in their drug-loaded or unloaded forms. Encapsulated LNZ preserved its antimicrobial activity, displaying Minimal Inhibitory and Bactericidal Concentrations (MIC and MBC) of 2 mu g/mL and 16 mu g/mL, respectively, against S. aureus, MRSA, S. epidermidis and Enterococcus faecium. Notably, sub-MIC concentrations of PLA-PEG-NAC@LNZ NPs reduced MRSA biofilm formation more effectively than free LNZ as demonstrated by biomass inhibition (62% vs 39%) and fluorescence microscopy. At 8 & times; MIC, PLA-PEG-NAC@LNZ NPs demonstrated activity against preformed MRSA biofilm either in the early (49% biofilm reduction) and late stages (30-41% biofilm reduction, depending on exposure time). Furthermore, the combined use of PLA-PEG-NAC@LNZ NPs and PLA-PEG-NAC@Pent NPs produced an additive antibacterial effect against Escherichia coli supporting the potential of Pent to sensitize Gram-negative bacteria to Gram-positive-targeting antibiotics such as LNZ.

Ultrasmall silver nanoclusters have emerged as promising tools for their sensing, optical, and biological properties, but their translation toward practical applications remains limited due to the synthetic complexity and insufficient control over colloidal stability. Here we report a straightforward one-step strategy for the direct conversion of Ag+ ions into blue-emitting silver nanoclusters (FcCAR@Ag NCs, average diameter 2.6 nm) using amphiphilic ferrocene carnosine (FcCAR) as both reducing and capping agents. Under mild basic conditions and in the presence of ascorbic acid, the FcCAR ligand drives the kinetic-controlled reduction of silver ions and simultaneously stabilizes the resulting nanoclusters through interfacial interactions mediated by ferrocene and carnosine functional groups. The formation of well-dispersed nanoclusters is confirmed by comprehensive optical, structural, and colloidal characterization, revealing blue photoluminescence and high colloidal stability. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) analyses performed on screen-printed electrodes (SPEs) showed a higher electrochemical response of FcCAR@Ag NCs with respect to the native FcCAR ligand, suggesting the potential application of FcCAR@Ag NCs in electrochemical sensing. Moreover, the use of ligands based on peptides functionalized with ferrocene units introducing lipophilicity confers an amphiphilic character to NCs, which became crucial for the effective interaction with bacterial envelopes. Therefore, MIC and MBC values of FcCAR@Ag NCs against Staphylococcus aureus and Escherichia coli demonstrated the superior antimicrobial efficacy of silver in nanocluster form compared with conventional silver ions (AgNO3). Specifically, a two-fold reduction of MIC (from 11.7 to 5.8 µg mL-1 for S. aureus and from 5.8 to 2.9 µg mL-1 for E. coli) along with a four-fold reduction in MBC (from 46.7 to 11.7 µg mL-1 for S. aureus and from 11.7 to 2.9 µg mL-1 for E. coli) was observed. In addition, low MIC values (5.8 µg mL-1) were found against clinically relevant bacteria, including methicillin-resistant S. aureus (MRSA), vancomycin-resistant Enterococcus faecium (VREfm), Pseudomonas aeruginosa, and ESBLs producing Escherichia coli. Moreover, FcCAR@Ag NCs were also effective in reducing biomass and metabolic activity of 24 h-established biofilms formed by S. aureus, E. coli, and P. aeruginosa strains. Overall, our findings highlight the strong antimicrobial potential of FcCAR@Ag NCs against both Gram-positive and Gram-negative bacteria, including antibiotic-resistant and biofilm-producing strains.

Ataxia-Telangiectasia (AT) is a rare disorder characterized by ATM deficiency and mitochondrial dysfunction. Because SIRT3 contributes to mitochondrial homeostasis in ATM-deficient settings, it represents a relevant molecular context for repurposing-oriented target-engagement studies. Here, we present an integrated computational and biophysical workflow designed to identify experimentally tractable SIRT3 binders within a clinically approved chemical space. A curated dataset of 2342 FDA-approved drugs was screened using hierarchical GNINA-based virtual screening, SAveRUNNER network analysis, triplicate 300 ns molecular dynamics simulations, post-MD redocking, MM/PBSA calculations, and experimental SPR validation. Four compounds were prioritized for further investigation: binimetinib, olaparib, mizolastine, and teniposide. Surface plasmon resonance (SPR) demonstrated direct binding of all four compounds to recombinant SIRT3, with equilibrium dissociation constants in the nano- to low-micromolar range. Binimetinib showed the highest apparent affinity (KD = 21.28 ± 3.5 nM), followed by olaparib (84 ± 5.7 nM), mizolastine (138 ± 3.5 nM), and teniposide (1470 ± 58 nM). Computational analyses supported the compatibility of these ligands with dynamically relaxed SIRT3 binding-site conformations and enabled the efficient prioritization of experimentally validated binders. These results support the value of an integrated in silico/SPR pipeline for identifying SIRT3-binding scaffolds. However, the functional consequences of binding, isoform selectivity, and biological relevance in ATM-deficient disease models remain to be established.

Ataxia-Telangiectasia (AT) is a rare disorder caused by ATM deficiency and characterized by genomic instability, mitochondrial dysfunction, and increased cancer susceptibility. Because SIRT3 contributes to mitochondrial homeostasis and metabolic adaptation in ATM-deficient settings, compounds that bind this target may provide useful starting points for repurposing-oriented therapeutic exploration. In this study, a curated dataset of 2,342 FDA-approved drugs was screened using an integrated workflow combining GNINA-based virtual screening, SAveRUNNER network analysis, 300 ns molecular dynamics simulations, post-MD redocking, MM/PBSA calculations, and experimental biophysical validation. Four compounds were prioritized for further investigation: binimetinib, olaparib, mizolastine, and teniposide. Surface plasmon resonance (SPR) demonstrated direct binding of all four compounds to recombinant SIRT3, with equilibrium dissociation constants in the nano- to submicromolar range. Binimetinib showed the highest apparent affinity (KD = 21.28 ±7 nM), followed by olaparib (84 ±8.1 nM), mizolastine (138 ±11 nM), and teniposide (811.25 ±5.3 nM). Computational analyses supported the compatibility of these ligands with dynamically relaxed SIRT3 binding-site conformations and enabled the efficient prioritization of experimentally tractable candidates. Overall, this study identifies four FDA-approved SIRT3-binding compounds and supports the value of an integrated computational/SPR workflow for repurposing-oriented hit identification in AT-related malignancy settings. Although these compounds displayed distinct translational profiles, the present results consistently indicated direct target engagement rather than definitive enzymatic inhibition, selectivity, or functional activity in disease-relevant models.

The integration of molecular imprinting technology with electrochemical methods has become fundamental in the development of next-generation sensors. This study explores two different strategies for developing a dopamine-based molecularly imprinted polymer (MIP) for the electrochemical sensing of levofloxacin. In the first case, the MIP is developed by electropolymerization on a screen-printed carbon electrode (SPCE) surface using cyclic voltammetry, while in the second, the MIP is obtained by an oxidation process, and the resulting dispersion is drop-cast on the SPCE surface. The same approach is used for a non-imprinted polymer. The physicochemical properties of the synthesized materials and the surface morphology of the modified electrodes are investigated by several techniques. Differential pulse voltammetry is used to evaluate the performance of the modified electrodes, assessing their linear concentration range, limit of detection, and limit of quantification, together with repeatability and selectivity. MIP-based SPCEs obtained with these two fabrication strategies exhibited comparable imprinting factor values and linear concentration ranges, along with comparable limits of detection and quantification. The MIP-based SPCE obtained by electropolymerization showed greater repeatability, whereas the MIP-based SPCE produced by drop-casting provided higher sensitivity in levofloxacin detection.

This review provides a comprehensive and critical overview of MagnetoCyclodextrins (MNP-CDs), an emerging class of hybrid nanomaterials that merge the unique magnetic properties of iron oxide nanoparticles with the inclusion, encapsulation, and solubilization abilities of cyclodextrin derivatives. Specifically, we reviewed the synthetic strategies for their preparation and their potential applications in biomedical fields. The synthetic methods, including co-precipitation, chemical coating, phase transfer, and self-assembly, are discussed in detail, with a focus on their impact on colloidal stability, biocompatibility, surface functionality, and magnetic properties. Despite the encouraging in vitro and in vivo results, the translation of MNP-CDs into clinical practice is still at an early stage, and the relationship between their magnetic characteristics and therapeutic efficacy remains poorly explored. Future research should address these limitations by optimizing synthesis protocols for scalability and reproducibility and by developing targeted surface modifications. Accordingly, this review offers a comprehensive perspective on MNP-CDs fabrication methods, emphasizing the impact of different ligands, polymers, and inorganic components on their synthesis and functionalization. In addition, a critical assessment of the key challenges that must be addressed for their successful application in advanced biomedical fields is provided.

Osteosarcoma (OS) is a malignant bone tumor primarily affecting children and teenagers, characterized by aggressiveness and early metastasis especially to the lungs. OS management is complex and combined-modality therapy involving surgery, chemotherapy and immunotherapy is common. The standard care treatment utilizing doxorubicin, cisplatin, and high-dose methotrexate is a combination ("MAP") not changed in more than 40 years that often confronts incomplete tumor removal, recurrence, drug resistance, and severe side effects. Recent advancements in nano- and precision medicine have introduced tumor-targeted drug delivery strategies through multifunctional nanocarriers which aim to enhance therapeutic efficacy by preventing rapid clearance, prolonging circulation time and improving accumulation at tumor sites while minimizing adverse effects. Although many of these smart Nanotherapeutics are still at the preclinical stage, their unique properties make their promotion in OS clinical applications a challenge. Starting from an overview of the current approved OS therapies, this review reports a systematic analysis of in vivo studies published in the last decade that employ multifunctional nanosystems, drug delivery strategies and cutting-edge technologies in chemo-, immuno- and gene therapy for OS management providing an overview of the potential and challenges of these innovative treatment strategies. Our comprehensive literature analysis points out their certain antitumoral effects in OS preclinical models; however, overcoming translational bottlenecks remains a critical challenge, as promising preclinical findings often fail to translate into effective clinical therapies. Moreover, extended long-term observation in clinical studies is still required together with an in-depth understanding of the unique genetics and biology of OS, given the complex heterogeneity of the tumor microenvironment. By analyzing the limitations of conventional therapies, the latest advancements in nanotechnology alongside key bottlenecks in clinical translation of nanotherapeutics for OS, this review provides valuable insight into future directions, particularly for combination regimens, fostering progress in OS clinical research and supporting the development of innovative and personalized therapies.

Modifying an SPCE with graphene-cyclodextrin/ferrocenyl-carnosine supramolecular assembly improves the electrochemical detection of Mn( ii ), resulting in increased sensitivity and low detection limit.

This study presents two new hybrid nanosystems (G-PMA(1 : 1)@AuBPs and G-PMA(1 : 3)@AuBPs), constructed from amine graphene (G-NH2) functionalized with poly(methacrylic acid) (PMA) and gold nanoparticles with a bipyramidal shape (AuBPs). These nanoplatforms behave like efficient photothermal agents, making them suitable for effective in vitro photothermal therapy and for bioimaging applications simultaneously. The nanosystems were synthesized by combining covalent and supramolecular approaches and characterized by several techniques including XPS, Raman spectroscopy, UV-vis spectroscopy, XRD, and STEM. It was observed that G-PMA@AuBP systems demonstrate remarkable light-to-heat conversion efficiency under near-infrared irradiation at 785 and 808 nm. Both systems showed an enhancement of the photothermal properties compared to the individual materials. Particularly, a photothermal conversion efficiency exceeding 70% was estimated for the G-PMA(1 : 3)@AuBP sample under 808 nm irradiation. Beyond their photothermal capabilities, G-PMA@AuBP systems can be effective as label-free bioimaging probes. G-PMA(1 : 1)@AuBP has been successfully visualized within B16F10 melanoma cells using FLIM, conventional fluorescence, and dark-field microscopy techniques, with localization observed in the perinuclear region. Cytotoxicity assays confirmed the biocompatibility of both nanosystems. Finally, the in vitro phototherapeutic efficacy was validated under 808 nm laser irradiation, showing promising results for melanoma cell treatment through photothermal therapy.

This review provides a comprehensive overview of the therapeutic potential of omaveloxone (OMA) for the treatment of Friedreich’s ataxia (FA), along with an analysis of the historical development and current status of the synthetic strategies for OMA production. OMA activates the nuclear factor-2-(erythroid-2)-related (Nrf2) pathway in vitro and in vivo, in both animal models and humans. The Nrf2 pathway plays a crucial role in the cellular response to oxidative stress. Furthermore, OMA has been shown to mitigate mitochondrial dysfunction, restore redox homeostasis and downregulate nuclear factor-κB (NF-κB), a key mediator of inflammatory responses. Through these mechanisms, OMA contributes to tissue protection and inflammation reduction in patients with FA. The review also highlights future perspective, focusing on the challenges associated with OMA reprofiling through innovative drug delivery approaches and its potential repurposing for diseases beyond FA.

Skeletal muscle integrity and its intrinsic aligned architecture are crucial for locomotion, postural support, and respiration functions, impacting overall quality of life. However, volumetric muscle loss (VML) can exceed intrinsic regenerative potential, leading to fibrosis and impairments. Autologous muscle grafting, the current gold standard, is constrained by tissue availability and success rates. Therefore, innovative strategies like cell-based therapies and scaffold-based approaches are needed. Our minimally invasive approach involves a tunable injectable hydrogel capable of achieving an aligned architecture post-injection via a low-intensity static magnetic field (SMF).Our hydrogel formulation uses gellan gum as the backbone polymer, enriched with essential extracellular matrix components such as hyaluronic acid and collagen type I, enhancing bio-functionality. To achieve an aligned architectural biomimicry, collagen type I is coupled with iron oxide magnetic nanoparticles, creating magnetic collagen bundles (MagC) that align within the hydrogel when exposed to a SMF. An extensive study was performed to characterize MagC and assess the hydrogel’s stability, mechanical properties, and biological response in vitro and in vivo.The proposed system, fully composed of natural polymers, exhibited mechanical properties similar to human skeletal muscle and demonstrated effective biological performances, supporting its potential as a safe and patient-friendly treatment for VML.

Neutron detectors are paramount in many applications from medical science to homeland security and aero-space. A miniaturized solid-state device based on a nanostructured-gold thin film grown by pulsed laser ablation under deposition-controlled conditions and functionalized with a monolayer of 4-mercaptophenyboronic acid (4-MPBA) was fabricated as a neutron dose detector. The device is tested with slow neutron detection in the thermal and epithermal energy range at ISIS spallation neutron source (UK). After the neutron irradiation, 4-MPBA is converted into thiophenol (TP), and the chemical modification is monitored by the intensity of related vibra-tional bands at 1586 cm-1 (4-MPBA) and 1574 cm-1 (TP). The latter are used as a vibrational signature of the absorbed dose by surface-enhanced Raman spectroscopy (SERS). The conversion of 4-MPBA to TP is due to the loss of the boron-containing group after the absorption of a slow neutron by 10B isotope. The I1574/I1586 ratio is used to estimate the ratio of 10B nuclei absorption reactions due to the thermal and epithermal contributions and it is proposed for the development of neutron dosimetry for nuclear medicine, aerospace, and security applications.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has provoked a global health crisis due to the absence of a specific therapeutic agent. 3CLpro (also known as the main protease or Mpro) and PLpro are chymotrypsin-like proteases encoded by the SARS-CoV-2 genome, and play essential roles during the virus lifecycle. Therefore, they are recognized as a prospective therapeutic target in drug discovery against SARS-CoV-2 infection. Thus, this work aims to collectively present potential natural 3CLpro and PLpro inhibitors by in silico simulations and in vitro entry pseudotype-entry models. We screened luteolin-7-O-glucuronide (L7OG), cynarin (CY), folic acid (FA), and rosmarinic acid (RA) molecules against PLpro and 3CLpro through a luminogenic substrate assay. We only reported moderate inhibitory activity on the recombinant 3CLpro and PLpro by L7OG and FA. Afterward, the entry inhibitory activity of L7OG and FA was tested in cell lines transduced with the two different SARS-CoV-2 pseudotypes harboring alpha (α) and omicron (o) spike (S) protein. The results showed that both compounds have a consistent inhibitory activity on the entry for both variants. However, L7OG showed a greater degree of entry inhibition against α-SARS-CoV-2. Molecular modeling studies were used to determine the inhibitory mechanism of the candidate molecules by focusing on their interactions with residues recognized by the protease active site and receptor-binding domain (RBD) of spike SARS-CoV-2. This work allowed us to identify the binding sites of FA and L7OG within the RBD domain in the alpha and omicron variants, demonstrating how FA is active in both variants. We have confidence that future in vivo studies testing the safety and effectiveness of these natural compounds are warranted, given that they are effective against a variant of concerns.

The growing interest in Kv7.2/7.3 agonists originates from the involvement of these channels in several brain hyperexcitability disorders. In particular, Kv7.2/7.3 mutants have been clearly associated with epileptic encephalopathies (DEEs) as well as with a spectrum of focal epilepsy disorders, often associated with developmental plateauing or regression. Nevertheless, there is a lack of available therapeutic options, considering that retigabine, the only molecule used in clinic as a broad-spectrum Kv7 agonist, has been withdrawn from the market in late 2016. This is why several efforts have been made both by both academia and industry in the search for suitable chemotypes acting as Kv7.2/7.3 agonists. In this context, in silico methods have played a major role, since the precise structures of different Kv7 homotetramers have been only recently disclosed. In the present review, the computational methods used for the design of Kv.7.2/7.3 small molecule agonists and the underlying medicinal chemistry are discussed in the context of their biological and structure-function properties.

A consistent series of Pt (II) polypyridyl complexes (i.e., LDP‐10–25 ), previously obtained and characterized by our research group, underwent extensive biological investigations to verify their activity profile as target‐based anticancer agents. Preliminary in vitro screening at 10 μM against three tumor cell lines known to overexpress DNA G‐4 (MDA‐MB 231, U87, and U2‐OS) pointed out that four of them, namely, LDP‐15 , LDP‐16 , LDP‐24 , and LDP‐25 , had promising cytotoxic activity compared with cisplatin. Therefore, these four compounds were selected for continuous assays against the same three cell lines and morphological analyses on U2‐OS cells that showed IC 50 values in the micromolar range and remarkable changes in nuclei shape and cytoskeleton integrity, respectively. Docking studies supported the idea that the antiproliferative activity of the complexes could be attributed to their interaction via a hybrid binding mode with the intended molecular target, DNA G‐4. In addition, in silico ADME‐Tox profiling studies showed no risk of tumorigenic, irritant, or reproductive effects for the title compounds. DFT calculations were used to verify the structural characteristics of the four selected compounds and to investigate their electronic behavior. Overall, the results obtained, both experimentally and theoretically, indicate that LDP‐15 , LDP‐16 , LDP‐24 , and LDP‐25 complexes could be useful for further study as potential therapeutic agents.

Cellular alignment plays a pivotal role in several human tissues, including skeletal muscle, spinal cord and tendon. Various techniques have been developed to control cellular alignment using 3D biomaterials. However, the majority of 3D-aligned scaffolds require invasive surgery for implantation. In contrast, injectable hydrogels provide a non-invasive delivery method, gaining considerable attention for the treatment of diverse conditions, including osteochondral lesions, volumetric muscle loss, and traumatic brain injury.We engineered a biomimetic hydrogel with magnetic responsiveness by combining gellan gum, hyaluronic acid, collagen, and magnetic nanoparticles (MNPs). Collagen type I was paired with MNPs to form magnetic collagen bundles (MCollB), allowing the orientation control of these bundles within the hydrogel matrix through the application of a remote low-intensity magnetic field. This resulted in the creation of an anisotropic architecture. The hydrogel mechanical properties were comparable to those of human soft tissues, such as skeletal muscle, and proof of the aligned hydrogel concept was demonstrated.In vitro findings confirmed the absence of toxicity and pro-inflammatory effects. Notably, an increased fibroblast cell proliferation and pro-regenerative activation of macrophages were observed. The in-vivo study further validated the hydrogel biocompatibility and demonstrated the feasibility of injection with rapid in situ gelation. Consequently, this magnetically controlled injectable hydrogel exhibits significant promise as a minimally invasive, rapid gelling and effective treatment for regenerating various aligned human tissues.