Search Results (332)

Low-temperature plasma treatments were applied to barley seeds using a dielectric barrier-stabilized corona discharge operated in ambient air enriched with oxygen or nitrogen to quantify surface chemical modifications and seed wettability. X-ray photoelectron spectroscopy showed that oxygen-enriched plasma produced the strongest oxidation, increasing surface oxygen from 9 ± 5 at% (control) to 24 ± 5 at%, while reducing carbon from 88 ± 5 at% to 76 ± 5 at%. Nitrogen-enriched plasma induced more moderate changes (O: 13 ± 5 at%, C: 85 ± 5 at%) but resulted in clear nitrogen incorporation, with an enhanced N 1s amine/amide component at ~400.8 eV. The hydroxyl O 1s contribution increased from 70% (control) to 82% (oxygen) and 90% (nitrogen), indicating substantial surface hydroxylation. SEM-EDX showed only minor micrometer-scale composition changes and no detectable morphological damage. Raman and ATR-FTIR spectra confirmed that polysaccharide, protein, and lipid structures remained intact, with intensity variations reflecting increased hydrophilicity. Water imbibition kinetics fitted with the Peleg model demonstrated faster initial hydration after plasma exposure, with 1/k₁ increasing from 20.25 ± 1.90 h⁻¹ (control) to 36.70 ± 6.56 h⁻¹ (oxygen) and 38.87 ± 7.57 h⁻¹ (nitrogen), while 1/k₂ remained nearly unchanged. Full article

GenX, also known as hexafluoroepoxypropane dimer acid (HFPO-DA), an emerging perfluoroalkyl substance alternative, is extensively used in industrial processes and is resistant to degradation. This persistence heightens the potential for co-occurrence and combined toxicity with other environmental pollutants. Nanoplastics (NPs), ubiquitous environmental contaminants, can exacerbate the biological toxicity of GenX. However, the molecular mechanisms by which NPs influence GenX-induced structural damage to human serum albumin (HSA) remain unclear. This study, therefore, employed multi-spectroscopic techniques, characterization assays, and molecular simulations to investigate these mechanisms. A critical limitation is that the observed structural damage occurred at a GenX concentration of 0.05–0.1 mM. The results indicate that the presence of NPs exacerbated the loosening of the protein backbone and caused a more pronounced reduction in α-helical content (NPs@GenX: 37.3%; GenX alone: 41.5%). The binding is predicted to occur within the hydrophobic pocket of subdomain IIIA of HSA. Characterization assays further revealed significant protein aggregation in systems containing NPs. The study concludes that NPs adsorb HSA through the formation of a protein corona, while simultaneously binding GenX via hydrophobic interactions. This dual pathway—direct binding of HSA to GenX and an active surface-mediated perturbation by NPs—constitutes the primary mechanism leading to aggravated structural changes. Overall, this work elucidates the molecular mechanisms by which NPs exacerbate HSA denaturation in the presence of GenX, offering valuable insights for assessing the combined ecological risks of emerging and persistent environmental pollutants. Full article

Exogenous insulin is essential for diabetes management; however, subcutaneous administration is associated with discomfort, poor adherence and non-physiological peripheral hyperinsulinemia. Oral administration would better mimic the physiological insulin distribution route but is hampered by gastrointestinal barriers, resulting in low bioavailability. Enabling access to the market for oral insulin nanocarriers requires rigorous control of their physicochemical attributes (size, charge, and surface chemistry) to ensure biocompatibility and mitigate risks such as the long-term bioaccumulation of non-biodegradable materials and the loss of intended targeting due to protein corona formation. In pre-clinical studies, nanoparticle carriers have shown promising results by protecting insulin and enhancing its absorption, yet clinical translation remains limited, with most candidates stalling in early-phase trials. This translational gap stems from the inadequacy of conventional animal models and regulatory frameworks to address the complexity of nanomedicines. This review goes beyond a simple summary of nanocarrier types and discusses the non-clinical and regulatory challenges hampering progress. We highlight the limitations of current preclinical models and the challenge of evaluating the pharmacokinetic profiles of both the nanocarrier and its insulin payload. The development of more rigorous and predictive strategies based on most recent successes and failures, described in this review, could help to bridge the translational gap. Full article

Background/Objectives: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can naturally infect a broad spectrum of animal species, with cats, minks, and ferrets being highly susceptible. There is a potential risk that infected animals could transmit viruses to humans. Moreover, SARS-CoV-2 continues to evolve via mutation and genetic recombination, resulting in the continuous emergence of new variants that have triggered a wave of reinfection. Therefore, safe and effective corona virus disease 2019 (COVID-19) vaccines for animals are still being sought. Methods: We generated three recombinant Modified vaccinia virus Ankara (MVAs) expressing the prefusion-stabilized S proteins, S6P, DS6P, and BA2S6P, targeting the full-length S protein genes of the ancestral, Delta, and Omicron BA.2 strains of SARS-CoV-2. Subsequently, the safety, immunogenicity, and protective efficacy of these MVA-based oral COVID-19 vaccine candidates were assessed in mice, minks, and cats. Results: These recombinant MVAs are safe in mice, minks, and cats. Oral or intramuscular vaccination with rMVA-S6P induced a robust SARS-CoV-2 neutralizing antibody (NA) response and conferred complete protection against the SARS-CoV-2 challenge in mice. Meanwhile, oral or intramuscular administration of these recombinant MVAs in combination induced a potent and durable NA response against homotypic SARS-CoV-2 pseudovirus in mice, minks, and cats, respectively. Conclusions: These findings suggest that the MVA-vectored vaccines are promising oral COVID-19 vaccine candidates for animals, and that the combined vaccination approach is an effective administration strategy for such vaccines. Full article

This review provides a comprehensive overview of the fundamental aspects of nanoparticles (NPs), emphasizing their physicochemical properties and biological interactions, with particular focus on porous silica nanoparticles (PSNs). The review provides information on the Safe-by-design (SbD) S.A.F.E. (Standardised characterization, Assessment of biocompatibility, Facilitation of toxicity and exposure routes and Evaluation of clinical translation) framework. It discusses critical factors influencing NP toxicity and cellular uptake, including particle size, shape, pore size, surface charge, surface functionalisation, and crystallinity. The review also examines exposure routes of NPs—inhalation, dermal, oral, systemic and mucosal—and their subsequent biological effects. A key section is dedicated to the formation of the protein corona, a critical determinant of NP fate in biological systems, and its influence on circulation time, immune clearance and cellular responses. Particular attention is given to assessing the biological interactions of the PSNs and the mechanisms underlying PSN-induced cytotoxicity and genotoxicity, with a focus on the assays commonly employed to evaluate these effects. The review explores the use of gene expression profiling as a powerful tool to elucidate the molecular mechanisms underlying nanoparticle-induced cellular changes. This review aims to provide an integrated perspective on the SbD considerations and safety implications of nanomaterials. It highlights the need for a deeper understanding of complex biological interactions to establish SbD principles and enable the translation of PSNs into clinical applications. Finally, current regulatory frameworks and guidelines for testing nanomaterials, including PSNs, that support their safe and sustainable development are discussed. Full article

Chronic and hard-to-heal wounds remain burdensome because microbial contamination, dysregulated inflammation, and fragile tissue regeneration slow closure, while passive dressings often injure new tissue during removal. This review synthesizes polymer- and lipid-based nanostructures through a barrier-resolved lens that links composition, architecture, and processing to performance in protease- and salt-rich exudate across topical and transdermal routes. Quantitative trends include effective diameters of approximately 50–300 nm, practical constraints of sterile filtration at 0.2 μm, and therapeutic windows that prioritize contamination control on the first day, support proliferation around day three, and sustain remodeling beyond one week. Mechanistic evidence indicates that interfacial charge and the protein corona govern residence and uptake, lipid bilayers enable dual loading, degradable polymer matrices provide depot-like behavior, and hybrid constructs temper the early burst while improving storage stability. Full article

For complexation of mRNA into polyplexes, double-pH-responsive lipo-xenopeptides (XP), comprising tetraethylene pentamino succinic acid (Stp) and lipoamino fatty acids (LAFs), were combined with PEGylated lipids, either DMG-PEG 2 kDa (DMG-PEG) or azido-group-containing DSPE-PEG 2 kDa (DSPE-PEG-N3), to increase colloidal stability and to facilitate ligand-mediated targeted mRNA delivery. LAF-XPs mixed with DMG-PEG at low (1.5% and 3%) molar ratios improved colloidal stability and retained transfection efficiency. PEGylation also enabled the formulation of otherwise unstable carrier complexes and prevented aggregation induced by salt, proteins, and serum. PEGylation of more positively charged Stp-LAF₂ mRNA polyplexes decreased fibrinogen adsorption. More neutral, LAF-rich Stp-LAF₄ polyplexes exhibited low fibrinogen binding without PEGylation. Intravenous administration of these stabilized mRNA complexes demonstrated enhanced biosafety while preserving transfection efficiency. DSPE-PEG-N3 was selected for cell targeting after strain-promoted azide-alkyne cycloaddition (SPAAC)-mediated click-coupling of DBCO-modified ligands. Higher PEG ratios (10% and 20%) provided effective shielding but reduced transfection efficiency, a drawback known as the “PEG dilemma”. Functionalization with an EGFR-targeting ligand restored transfection in EGFR-positive cell lines in a ligand-specific manner. High transfection efficiency is consistent with a lipophilic-to-hydrophilic polarity switch of LAF-XP carriers upon endosomal protonation, triggering dissociation of the PEG lipids and deshielding of the polyplex. Full article

Background: Mesoporous silica nanoparticles (MS NPs) have attracted significant interest for their role in the advancement of drug delivery systems. However, further investigation is needed to unravel the mechanisms behind the shift in organ tropism that occurs with changes in composition. Methods: To shed light on the correlation between their composition and organ-targeting capabilities, a range of MS NPs was synthesized and subsequently administered intravenously to mice. Results: Our results indicate that MS NPs with a pristine -Si-O-Si- framework, or those incorporating -C-C- or –S-S-S-S- bonds, predominantly accumulated in the liver. The shift to lung tropism was observed exclusively in MS NPs that were enriched with –SH groups. Proteomic analysis identified histidine-rich glycoprotein (HRG) as the most prevalent protein associated with liver-preferred MS NPs in serum, while lung-preferred MS NPs, such as thioether-bridged deformable hollow mesoporous organosilica nanoparticles (HSMONs), showed the highest affinity for albumin. Furthermore, the lung-selective HSMONs, endowed with inherent deformability and glutathione-responsive biodegradability, were utilized as systemic nanocarriers for the delivery of gambogic acid (GA). Conclusions: Leveraging albumin absorbing-triggered tumor cell targeting and trafficking, HSMONs conjugated with GA effectively elicited potent antitumor effects in pulmonary tissue. Full article

Microplastics entering the gastrointestinal environment rapidly acquire protein coronas that alter their surface chemistry and analytical detectability. We investigated the physicochemical interactions between fluorescently labeled bovine serum albumin (BSA) and polyethylene terephthalate (PET) microplastics during simulated intestinal exposure and evaluated the stability of the resulting hard corona. Using fluorescence tracking, SDS-PAGE, and FTIR spectroscopy, we showed that BSA forms a persistent corona that resists oxidative-only treatments. Only a combination of oxidation with an alkaline (KOH) or surfactant step (SDS) effectively removed the corona. None of the protocols applied affected polymer integrity. Residual protein in less effective protocols did not show changes on PET spectra in ATR FTIR. To validate the protocol under physiologically relevant complexity, we extended it to PET incubated with single digestive enzymes. FTIR spectra confirmed the removal of protein-specific signals in both systems, with no degradation of PET ester or aromatic functional groups nor signals of protein–polymer interactions. Our results highlight the robustness of protein–PET interactions in biological conditions and provide a variety of protocols for protein corona removal, suitable for diverse applications of microplastic analysis and toxicological studies. Full article

Lipid nanoparticles are a clinically validated platform for delivering nucleic acids, but performance is constrained by multiscale physiological barriers spanning circulation, vascular interfaces, extracellular matrices, cellular uptake, and intracellular trafficking. This review links composition–structure–function relationships for ionizable lipids, helper phospholipids, cholesterol, and PEG-lipids to systemic fate, endothelial access, endosomal escape, cytoplasmic stability, and nuclear transport. We outline strategies for tissue and cell targeting, including hepatocyte ligands, immune and tumor selectivity, and selective organ targeting through compositional tuning, together with approaches that modulate escape using pH-responsive chemistries or fusion-active peptides and polymers. We further examine immunomodulatory co-formulation, route and schedule effects on biodistribution and immune programming, and manufacturing and stability levers from microfluidic mixing to lyophilization. Across these themes, we weigh trade-offs between stealth and engagement, potency and tolerability, and potency and manufacturability, noting that only a small fraction of endosomes supports productive release and that protein corona variability and repeat dosing can reshape tropism and clearance. Convergence of standardized assays for true cytosolic delivery, biomarker-guided patient selection, and robust process controls will be required to extend LNP therapeutics beyond the liver while sustaining safety, access, and scale. Full article

Currently, there is a growing interest in biomedical research in the use of inorganic nanoparticles for targeted drug delivery, as biosensors, and in theranostic applications. This review examines the interaction of inorganic nanoparticles with protein molecules depending on the chemical nature, size, and surface charge of the nanoparticles. The effect of protein and nanoparticle concentration, as well as their incubation time, is analyzed. The work focuses on the influence of parameters such as pH, ionic strength, and temperature on the interaction of nanoparticles with protein molecules. The following dependencies were studied in detail: the thickness of the protein corona as a function of nanoparticle size; the size of nanoparticles after interaction with protein as a function of protein and nanoparticle concentration; the distribution of zeta potentials in colloids of nanoparticles, proteins, and their mixtures. It has been shown that proteins and nanoparticles can influence each other’s physicochemical properties. This can lead to the emergence of new biological properties in the system. Therefore, the adsorption of proteins onto nanoparticle surfaces can induce conformational changes. The probability of changing the protein structure increases when a covalent bond is formed between the nanoparticle and the protein molecule. Studies demonstrate that protein structure remains more stable with spherical nanoparticles than with rod-shaped or other high-curvature nanostructures. The results presented in the review demonstrate the possibility of adapting physiological responses to nanomaterials by changing the chemical composition of the surface of nanoparticles and their size and charge. Full article

This brief review is a non-comprehensive look at some of the important aspects of lipidic nucleic acid delivery systems with a focus on RNA. In the context of this review on lipid-based formulation, nucleic acids are one of the key cargoes. Here, a brief historical background is given, highlighting a few of the key newly developing approaches to aid formulation design. These new techniques are discussed within a framework of “bottom-up” (rational) versus “top-down” (combinatorial) design. Evolving areas of interest that are discussed include multiplexed formulation and efficacy testing, new principles established in the role of the protein corona, details of the biophysical mechanism of delivery and machine learning approaches to design. Full article

When iron oxide nanoparticles are incubated together with a biological broth, the biomolecules compete for the binding sites at the solid–liquid interface. At the same time, the biomass rearranges in suspension, building agglomerated structures. Despite general knowledge of the forces involved in bio–nano interactions, gaps remain in the understanding of how biomolecules organize themselves in solution and onto surfaces. This work examines biomolecule adsorption onto metal oxide surfaces with the goal of strengthening this understanding, essential in industrial and natural processes. We demonstrate nearly complete separation of proteins from a biotechnological suspension for non-oxidized and highly oxidized magnetic nanoparticles. Varying the nanoparticle-to-biomass ratio, we find, can lead to different separation patterns, i.e., that selectivity using bare, low-cost materials is possible. Furthermore, we explore how preliminary “passivation” with a biological corona only partially reduces the ability to separate total protein mass from a new suspension in subsequent incubation steps. The study underscores the crucial role of concentration gradients with regard to targets and binding sites as the primary determinant of separation capacity and of biomolecule behavior in solution, highlighting the potential for using bio–nano coronae as biomolecule carriers across diverse fields, including environmental, biomedical, pharmaceutical and nutritional applications. Full article

In this study, we have established that unique composite particles (MLNCs) carried siRNA on a gold core and were covered with a lipid shell. MLNCs successfully delivered siRNa into cells in the presence of serum. We developed the photofixation method, allowing us to obtain MLNCs bearing a fixed protein corona. To understand the mechanisms of the influence that the protein corona has on the interaction of particles with cells, it is necessary to study the interaction of “naked” MLNCs with cells. This study aimed to examine the pathways of MLNC penetration into SC-1 fibroblasts used to confirm the efficacy of siRNA delivery. We studied fibroblasts in monolayer and spheroid form, and citrate AuNPs were used as a comparison particle. The same particles served as cores for MLNCs. The obtained results showed active penetration by clathrin-mediated endocytosis of “naked” MLNCs into SC-1 fibroblasts, regardless of the form of cultivation. AuNPs penetrated into monolayer fibroblasts by macropinocytosis and into spheroids by clathrin-mediated endocytosis. The penetration depth into the spheroids was about 40 μm for both types of particles (spheroid size was 350–400 μm). The particles migrated through the intercellular spaces, passing through intercellular contacts. Full article

This study examines the toxicological effects of carbon nanotubes (CNTs) of different diameters—single-walled CNTs (SWCNT, 2 nm) and multi-walled CNTs (MWCNT, 74 nm)—on two macrophage cell lines, rat alveolar NR8383 cells and human differentiated THP-1. Using standardized exposure conditions and employing an integrated omics approach (transcriptomic and proteomic analyses), both CNT types were found to induce cellular stress responses and inflammation, especially in NR8383 cells, with notable involvement of the Sirtuin signaling pathway. After 24 h, MWCNTs uniquely disrupted lipid metabolism in NR8383 cells, resulting in foam cell formation and syncytia. While SWCNTs were less disruptive to metabolic pathways, they significantly altered gene regulation, particularly RNA splicing mechanisms. The dispersion medium—fetal bovine serum (FBS) versus human surfactant—also modulated the observed toxicological responses, highlighting the critical role of the protein corona in influencing CNT-cell interactions. These findings demonstrate that CNT diameter significantly affects cytotoxicity and cellular response pathways in a cell-type-specific manner. Full article