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27 pages, 2758 KiB  
Review
A Review of SAW-Based Micro- and Nanoparticle Manipulation in Microfluidics
by Débora Amorim, Patrícia C. Sousa, Carlos Abreu and Susana O. Catarino
Sensors 2025, 25(5), 1577; https://doi.org/10.3390/s25051577 - 4 Mar 2025
Abstract
Surface acoustic wave (SAW)-based microfluidics has emerged as a promising technology for precisely manipulating particles and cells at the micro- and nanoscales. Acoustofluidic devices offer advantages such as low energy consumption, high throughput, and label-free operation, making them suitable for particle manipulation tasks [...] Read more.
Surface acoustic wave (SAW)-based microfluidics has emerged as a promising technology for precisely manipulating particles and cells at the micro- and nanoscales. Acoustofluidic devices offer advantages such as low energy consumption, high throughput, and label-free operation, making them suitable for particle manipulation tasks including pumping, mixing, sorting, and separation. In this review, we provide an overview and discussion of recent advancements in SAW-based microfluidic devices for micro- and nanoparticle manipulation. Through a thorough investigation of the literature, we explore interdigitated transducer designs, materials, fabrication techniques, microfluidic channel properties, and SAW operational modes of acoustofluidic devices. SAW-based actuators are mainly based on lithium niobate piezoelectric transducers, with a plethora of wavelengths, microfluidic dimensions, and transducer configurations, applied for different fluid manipulation methods: mixing, sorting, and separation. We observed the accuracy of particle sorting across different size ranges and discussed different alternative device configurations to enhance sensitivity. Additionally, the collected data show the successful implementation of SAW devices in real-world applications in medical diagnostics and environmental monitoring. By critically analyzing different approaches, we identified common trends, challenges, and potential areas for improvement in SAW-based microfluidics. Furthermore, we discuss the current state-of-the-art and opportunities for further research and development in this field. Full article
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Figure 1

Figure 1
<p>Example of a SAW-based microfluidic device for cell sorting. Adapted from [<a href="#B11-sensors-25-01577" class="html-bibr">11</a>], PNAS, under a Creative Commons Attribution (CC BY) license.</p>
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<p>(<b>a</b>) Schematics of an interdigitated transducer (IDT), showing the relation between the finger pairs and the wavelength of the resulting surface acoustic wave (SAW). (<b>b</b>) A typical SAW device, with the IDT patterned on the surface of the piezoelectric substrate. Reproduced from [<a href="#B30-sensors-25-01577" class="html-bibr">30</a>] with permission from Nature.</p>
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<p>Scheme of different IDT designs: (<b>a</b>) single electrode; (<b>b</b>) floating electrode; (<b>c</b>) chirped; (<b>d</b>) slanted; (<b>e</b>) focused.</p>
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<p>Schematic representation of a cross-sectional view of the two operation modes of SAW microfluidic devices. (<b>a</b>) Traveling SAW operation mode. (<b>b</b>) Standing SAW operation mode. Adapted from [<a href="#B23-sensors-25-01577" class="html-bibr">23</a>] with permission from Nature.</p>
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<p>Manipulation of fluorescent polystyrene beads. (<b>a</b>) 1D manipulation; (<b>b</b>) 2D manipulation; (<b>c</b>) microbead experimental patterning (1D); (<b>d</b>) microbead experimental patterning (2D). Reprinted with permission from [<a href="#B41-sensors-25-01577" class="html-bibr">41</a>] Copyright 2009, Royal Society of Chemistry.</p>
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<p>Schematic of a cross-sectional view of the SSAW microfluidic device, highlighting the forces acting within the system. The substrate generates Rayleigh waves, which induce both longitudinal and transverse motions. These motions give rise to acoustic streaming (depicted by red arrows) and the acoustic radiation force (<math display="inline"><semantics> <mrow> <msub> <mrow> <mi>F</mi> </mrow> <mrow> <mi>R</mi> </mrow> </msub> </mrow> </semantics></math> represented by green arrows). Blue dots indicate the pressure nodes, where the acoustic pressure amplitude is minimal, while red dots represent the pressure antinodes, where the acoustic pressure amplitude is maximal. Adapted from [<a href="#B59-sensors-25-01577" class="html-bibr">59</a>], PNAS, under a Creative Commons Attribution (CC BY) license.</p>
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<p>Schematic representation of different configurations of SAW-based microfluidic devices. (<b>a</b>) A SSAW-based device, for separation, with single electrode IDTs and a microfluidic channel with 3 inlets and 3 outlets. Reprinted from [<a href="#B13-sensors-25-01577" class="html-bibr">13</a>], American Chemical Society, under a Creative Commons Attribution (CC BY) license. (<b>b</b>) A device using SSAWs for concentration and TSAWs for separation. TSAWs are generated alternately by the focused IDTs on both sides of the channel. Adapted from [<a href="#B10-sensors-25-01577" class="html-bibr">10</a>], with permission from Elsevier. (<b>c</b>) A tilted-angle SSAW device for separation. Adapted from [<a href="#B42-sensors-25-01577" class="html-bibr">42</a>], American Chemical Society, under a Creative Commons Attribution (CC BY) license. (<b>d</b>) A SSAW-based device for 3D manipulation with two pairs of single-electrode IDTs arranged orthogonally to each other. Adapted from [<a href="#B59-sensors-25-01577" class="html-bibr">59</a>], PNAS, under a Creative Commons Attribution (CC BY) license. (<b>e</b>) A device with two slanted IDTs to generate TSAWs and manipulate a static sample. Adapted from [<a href="#B16-sensors-25-01577" class="html-bibr">16</a>], American Association for the Advancement of Science, under a Creative Commons Attribution (CC BY) license.</p>
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16 pages, 9618 KiB  
Article
Copper Hexacyanoferrates Obtained via Flavocytochrome b2 Assistance: Characterization and Application
by Galina Gayda, Olha Demkiv, Nataliya Stasyuk, Halyna Klepach, Roman Serkiz, Faina Nakonechny, Mykhailo Gonchar and Marina Nisnevitch
Biosensors 2025, 15(3), 157; https://doi.org/10.3390/bios15030157 - 2 Mar 2025
Viewed by 151
Abstract
Artificial enzymes or nanozymes (NZs) are gaining significant attention in biotechnology due to their stability and cost-effectiveness. NZs can offer several advantages over natural enzymes, such as enhanced stability under harsh conditions, longer shelf life, and reduced production costs. The booming interest in [...] Read more.
Artificial enzymes or nanozymes (NZs) are gaining significant attention in biotechnology due to their stability and cost-effectiveness. NZs can offer several advantages over natural enzymes, such as enhanced stability under harsh conditions, longer shelf life, and reduced production costs. The booming interest in NZs is likely to continue as their potential applications expand. In our previous studies, we reported the “green” synthesis of copper hexacyanoferrate (gCuHCF) using the oxidoreductase flavocytochrome b2 (Fcb2). Organic–inorganic micro-nanoparticles were characterized in detail, including their structure, composition, catalytic activity, and electron-mediator properties. An SEM analysis revealed that gCuHCF possesses a flower-like structure well-suited for concentrating and stabilizing Fcb2. As an effective peroxidase (PO) mimic, gCuHCF has been successfully employed for H2O2 detection in amperometric sensors and in several oxidase-based biosensors. In the current study, we demonstrated the uniqueness of gCuHCF that lies in its multifunctionality, serving as a PO mimic, a chemosensor for ammonium ions, a biosensor for L-lactate, and exhibiting perovskite-like properties. This exceptional ability of gCuHCF to enhance fluorescence under blue light irradiation is being reported for the first time. Using gCuHCF as a PO-like NZ, novel oxidase-based sensors were developed, including an optical biosensor for L-arginine analysis and electrochemical biosensors for methanol and glycerol determination. Thus, gCuHCF, synthesized via Fcb2, presents a promising platform for the development of amperometric and optical biosensors, bioreactors, biofuel cells, solar cells, and other advanced devices. The innovative approach of utilizing biocatalysts for nanoparticle synthesis highlights a groundbreaking direction in materials science and biotechnology. Full article
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Figure 1
<p>Examples of visualization in the ABTS- and <span class="html-italic">o</span>-dianisidine-based assays: (<b>a</b>)—PO-like activity of the chCuHCF (1) and gCuHCF (2–5) samples with the following activities (U/mL): 1—1.38; 2—2.12; 3—1.98; 4—3.78; 5—2.44. The substrate for PO-like activity contains a constant H<sub>2</sub>O<sub>2</sub> concentration. (<b>b</b>)—The dependence of color intensity on increasing H<sub>2</sub>O<sub>2</sub> concentrations (1–5), compared to the control sample without H<sub>2</sub>O<sub>2</sub> (6).</p>
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<p>Optical spectra of gCuHCF solutions (<b>a</b>) at different concentrations (mg/mL): 0 (1), 0.25 (2), 0.5 (3), 1 (4), 2 (5), and the calibration graph for photometric gCuHCF determination (<b>b</b>).</p>
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<p>Dependence of the optical density of the reaction mixture on H<sub>2</sub>O<sub>2</sub> concentration (<b>a</b>,<b>b</b>) and kinetic data linearization using the Lineweaver–Burke method (<b>c</b>). The initial gCuHCF concentration is 4.2 mg/mL.</p>
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<p>Examples of calibration graphs in the ArgO/gCuHCF/<span class="html-italic">o</span>-DZ naked-eye method for Arg determination (<b>a</b>), and the linearization of kinetic data using the Lineweaver–Burke method (<b>b</b>). The reaction mixtures (<b>a</b>) contain increasing concentrations of Arg (mM): 0 (1), 2 (2), 5 (3), 10 (4), 25 (5), 50 (6), 75 (7), 100 (8), with ArgO (1 U/mL) and varying gCuHCF concentrations (mg/mL): 4 (A), 2 (B), and 1 (C).</p>
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<p>Characteristics of the AO/gCuHCF/GE as an ABS for methanol: The CV profiles as outputs on methanol addition (<b>a</b>) up to concentration (mM): 0 (1, black), 2.5 (2, red) and 5 (3, blue), and the dependence of the current response on increasing concentrations of the analyte for a wide range (<b>b</b>) and a linear range (<b>c</b>). The GE was modified with 20 mU of gCuHCF exhibiting PO-like activity and 200 mU of AO. Conditions: scan rate (for <b>a</b>) is 50 mV·s<sup>−1</sup> vs. Ag/AgCl (reference electrode), working potential (for <b>b</b>,<b>c</b>) is 150 mV; 50 mM phosphate buffer, pH 7.0, 20 °C.</p>
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<p>Characteristics of the GlycO/gCuHCF/GE as an ABS for glycerol: The CV profiles as outputs on glycerol addition (<b>a</b>) up to concentration (mM): 0 (black), 12 (red), and 24 (blue); and the dependence of the current response on increasing concentrations of the analyte in the wide (<b>b</b>) and linear (<b>c</b>) ranges. GE was modified with 20 mU of gCuHCF exhibiting PO-like activity and 150 mU of GlycO. Conditions: scan rate (for <b>a</b>) is 50 mV s<sup>−1</sup> vs. Ag/AgCl (reference electrode), working potential (for <b>b</b>,<b>c</b>) is 150 mV; 50 mM phosphate buffer, pH 8.0, 20 °C.</p>
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<p>CV profiles of fresh-prepared gCuHCF/GE as biosensor on lactate: outputs on substrate addition up to concentration (mM): 0 (<b>1</b>, black), 15 (<b>2</b>, red), 30 (<b>3</b>, green), and 50 (<b>4</b>, blue). Conditions: scan rate 50 mV·s<sup>−1</sup> vs. Ag/AgCl as reference electrode, 50 mM acetate buffer, pH 6.0, 20 °C. The sensing layer contains 4.1 mU Fc<span class="html-italic">b</span><sub>2</sub>.</p>
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<p>Characteristics of the gCuHCF/Ges as ABS-1 and ABS-2 for lactate: the dependence of the current response on increasing concentrations of the analyte is demonstrated in both wide (<b>a</b>,<b>d</b>) and linear ranges (<b>b</b>,<b>c</b>,<b>e</b>,<b>f</b>). ABSs contain different amounts of fresh gCuHCFs: 5 µL (<b>1</b>, black line) and 10 µL (<b>2</b>, red line). Measurements were performed in 50 mM acetate buffer, pH 6.0 (<b>a</b>–<b>c</b>), and in 50 mM phosphate buffer, pH 7.0 (<b>d</b>–<b>f</b>), working potential of +300 vs. Ag/AgCl (reference electrode), at 23 °C mV.</p>
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<p>Characteristics of gCuHCF/GE as a chemosensor for ammonium ions: CV profiles as outputs on ammonium chloride addition (<b>a</b>) up to concentration (mM): 0 (<b>1</b>, black), 30 (<b>2</b>, red), and 60 (<b>3</b>, blue); chronoamperogram (<b>b</b>), dependence of the amperometric response on increasing ammonium chloride concentrations (<b>c</b>–<b>f</b>) in wide (<b>c</b>,<b>e</b>) and linear ranges (<b>d</b>,<b>f</b>). Conditions: scan rate (for <b>a</b>) is 50 mV·s<sup>−1</sup> vs. Ag/AgCl (reference electrode); temperature: 23 °C. Working potentials: +150 mV (<b>b</b>,<b>c</b>,<b>d</b>) and −100 mV (<b>e</b>,<b>f</b>). Buffers: 50 mM sodium acetate, pH 4.5 (<b>b</b>–<b>d</b>) and 50 mM sodium phosphate, pH 8.0 (<b>e</b>,<b>f</b>).</p>
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<p>Microscopic characterization of freshly prepared gCuHCFs suspended in 50 mM phosphate buffer: SEM image (<b>a</b>) and fluorescence images (<b>b</b>–<b>f</b>) captured under different filters. The images include brightfield (<b>b</b>) and blue-light fluorescence (<b>c</b>–<b>f</b>), showing dynamic changes over 1 to 5 min after irradiation with a 100 ms exposure.</p>
Full article ">Figure 10 Cont.
<p>Microscopic characterization of freshly prepared gCuHCFs suspended in 50 mM phosphate buffer: SEM image (<b>a</b>) and fluorescence images (<b>b</b>–<b>f</b>) captured under different filters. The images include brightfield (<b>b</b>) and blue-light fluorescence (<b>c</b>–<b>f</b>), showing dynamic changes over 1 to 5 min after irradiation with a 100 ms exposure.</p>
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<p>Microscopic characterization of gCuHCFs samples: SEM images (<b>a</b>,<b>e</b>) and fluorescence images (<b>b</b>–<b>d</b>,<b>f</b>–<b>h</b>) captured under different filters. The FM images include brightfield (<b>b</b>,<b>f</b>) and DAPI-stained fluorescence (<b>c</b>,<b>d</b>,<b>g</b>,<b>h</b>), showing dynamic changes over 1 to 5 min after irradiation with a 100 ms exposure. gCuHCF samples were kept for 1 month at 4 °C as a suspension in water (<b>a</b>–<b>d</b>) and as a lyophilized powder, which was suspended in water before characterization (<b>e</b>–<b>h</b>).</p>
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11 pages, 5035 KiB  
Article
Selective Carbon Dioxide Hydrogenation to Olefin-Rich Hydrocarbons by Cu/FeOx Nanoarchitectures Under Atmospheric Pressure
by Muhammad I. Qadir, Naděžda Žilková, Libor Kvítek and Stefan Vajda
Nanomaterials 2025, 15(5), 353; https://doi.org/10.3390/nano15050353 - 24 Feb 2025
Viewed by 271
Abstract
The conversion of carbon dioxide into fuels and fine chemicals is a highly desirable route for mitigating flue gas emissions. However, achieving selectivity toward olefins remains challenging and typically requires high temperatures and pressures. Herein, we address this challenge using 12 nm copper [...] Read more.
The conversion of carbon dioxide into fuels and fine chemicals is a highly desirable route for mitigating flue gas emissions. However, achieving selectivity toward olefins remains challenging and typically requires high temperatures and pressures. Herein, we address this challenge using 12 nm copper nanoparticles supported on FeOx micro-rods, which promote the selective hydrogenation of CO2 to light olefins (C2–C4) under atmospheric pressure. This catalyst achieves up to 27% conversion and 52% selectivity toward C2–C4 olefins, along with the production of C2–C4 paraffins, C5+ hydrocarbons (with all C1+ products totalling to up to about 75%), and methane, while suppressing CO formation to just 1% at 340 °C. The enhanced performance of the Cu/FeOx pre-catalyst is attributed to the efficient in situ generation of iron carbides (Fe5C2) in the presence of copper nanoparticles, as confirmed by ex situ XRD analysis. Copper facilitates the reduction of FeOx to form Fe5C2, a crucial intermediate for shifting the reaction equilibrium toward higher hydrocarbons. The hydrogenation of CO2 to higher hydrocarbons proceeds through the reverse water–gas shift reaction coupled with Fischer–Tropsch synthesis. Full article
(This article belongs to the Special Issue Nanocatalysts for Air Purification)
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Graphical abstract

Graphical abstract
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<p>(<b>a</b>,<b>b</b>) HR-TEM images of as-prepared Cu/FeOx; (<b>c</b>) STEM-HAADF of Cu/FeOx and EDS mapping of Fe, Cu and their overlap elements. The bars for EDS mapping images are 200 nm.</p>
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<p>SEM images of as-prepared Cu/FeOx catalyst (<b>a</b>–<b>c</b>), showing submicrometric structure of the FeOx rods and small nanoparticles of Cu (the orange rectangle with superscript 1 on (<b>c</b>)) on their surface. (<b>d</b>) EDX of the selected particles. The SEM images of the bare FeOx catalyst shown in <a href="#app1-nanomaterials-15-00353" class="html-app">Figure S1</a> reveal similar rods.</p>
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<p>XRD Rietveld refinement analysis of (<b>a</b>) as-prepared Cu/FeOx and (<b>b</b>) spent Cu/FeOx catalyst.</p>
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<p>Catalytic hydrogenation of CO<sub>2</sub> (<b>a</b>,<b>b</b>) by FeOx and (<b>c</b>,<b>d</b>) Cu/FeOx nanomaterials. Cat. 200 mg, CO<sub>2</sub>/H<sub>2</sub>/He (1/4/3.3, total flow 25 mL/min, reaction pressure 1 atm). After the first ramp, the catalyst was cooled down under He before starting the second ramp.</p>
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<p>(<b>a</b>,<b>b</b>) HR-TEM images of spent Cu/FeOx, and (<b>c</b>) STEM-HAADF and EDS mapping of Fe, Cu and C of the spent Cu/FeOx catalyst after 10 h. The bar in inset of Figure (<b>a</b>) is 20 nm, whereas the EDS mapping images have an 800 nm bar.</p>
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42 pages, 12382 KiB  
Review
Development of Wear-Resistant Polymeric Materials Using Fused Deposition Modelling (FDM) Technologies: A Review
by Zhiwang Li and Li Chang
Lubricants 2025, 13(3), 98; https://doi.org/10.3390/lubricants13030098 - 22 Feb 2025
Viewed by 486
Abstract
The advancement of 3D printing technology has changed material design and fabrication across various industries. Among its many applications, the development of high-wear-resistance polymer composites, particularly using Fused Deposition Modelling (FDM), has received increasing interest from both academic and industrial sectors. This paper [...] Read more.
The advancement of 3D printing technology has changed material design and fabrication across various industries. Among its many applications, the development of high-wear-resistance polymer composites, particularly using Fused Deposition Modelling (FDM), has received increasing interest from both academic and industrial sectors. This paper provides an overview of recent advances in this field, focusing on the selection of key printing parameters (such as layer thickness, print speed, infill density, and printing temperature) and material compatibility optimisation to enhance print quality and tribological performance. The effects of various tribo-fillers, such as fibres and nanoparticles, on the tribological properties of the printed polymer composites were studied. Generally, in the case of nano-sized particles, the wear rate can be reduced by approximately 3 to 5 times when the nanoparticle content is below 5 vol.%. However, when the nanoparticle concentration exceeds 10 vol.%, wear resistance may deteriorate due to the formation of agglomerates, which disrupts the uniform dispersion of reinforcements and weakens the composite structure. Similarly, in short fibre-reinforced polymer composites, a fibre content of 10–30 vol.% has been observed to result in a 3 to 10 times reduction in wear rate. Special attention is given to the synergistic effects of combining micro- and nano-sized fillers. These advancements introduce novel strategies for designing wear-resistant polymer composites without requiring filament fabrication, making 3D printing more accessible for tribological applications. In the last part of the review, the impact of emerging AI technologies on the field is also reviewed and discussed. By identifying key research gaps and future directions, this review aims to drive further innovation in the development of durable, high-performance materials for wide industry applications in aerospace, biomedical, and industrial engineering. Full article
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Figure 1

Figure 1
<p>Comparison of average wear depth for each print orientation (here, “horizontal”, “45° angle”, and “vertical” refer to the orientation of printed layers with respect to the sliding direction) [<a href="#B8-lubricants-13-00098" class="html-bibr">8</a>].</p>
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<p>Wear of ABS and PLA pins printed at three different raster angles and constant layer thicknesses of (<b>a</b>) 0.127 mm, (<b>b</b>) 0.254 mm, and (<b>c</b>) 0.32 mm [<a href="#B37-lubricants-13-00098" class="html-bibr">37</a>].</p>
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<p>Specific wear rate of FDM-printed PLA pins extruded at different extrusion temperatures [<a href="#B45-lubricants-13-00098" class="html-bibr">45</a>].</p>
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<p>Tribological performance of different volume fraction biocarbon-reinforced PLA [<a href="#B94-lubricants-13-00098" class="html-bibr">94</a>].</p>
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<p>Mechanical and wear properties in 3D-printed PLA silicon composites [<a href="#B9-lubricants-13-00098" class="html-bibr">9</a>].</p>
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<p>Typical evolutions of the COF and height loss as a function of the sliding time under different testing conditions: (<b>a</b>) COF with and without nanosilica taking in 0°-direction; (<b>b</b>) COF with and without nanosilica taking in 90°-direction; (<b>c</b>) height loss with and without nanosilica taking-in in 0°-direction and (<b>d</b>) height loss with and without nanosilica taking-in in 90°-direction [<a href="#B93-lubricants-13-00098" class="html-bibr">93</a>].</p>
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<p>EDX mappings of C- and Si-element of PEEK/CF30 after releasing PEEK/SiO220 in 90 direction. The sliding direction is from left to right [<a href="#B93-lubricants-13-00098" class="html-bibr">93</a>].</p>
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<p>Composition of printed PETG composites reinforced with SCFs and nano-clay particles [<a href="#B117-lubricants-13-00098" class="html-bibr">117</a>].</p>
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<p>Effect of load on the (<b>a</b>) SWR and (<b>b</b>) CoF of different PETG/SCF/OMMT nanocomposite samples (300 r/min, 500 m) [<a href="#B117-lubricants-13-00098" class="html-bibr">117</a>].</p>
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<p>Effect of sliding velocity on the (<b>a</b>) SWR and (<b>b</b>) CoF of different PETG/SCF/OMMT nanocomposite samples (25 N, 500 m) [<a href="#B117-lubricants-13-00098" class="html-bibr">117</a>].</p>
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<p>Comparison of experimental and numerical wear rates in the case of POM and glass fibre-reinforced (GFR) POM gear pairs [<a href="#B123-lubricants-13-00098" class="html-bibr">123</a>,<a href="#B124-lubricants-13-00098" class="html-bibr">124</a>].</p>
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<p>(<b>a</b>) Designed textured surface, (<b>b</b>) top-view of the printed textured surface of neat nylon, and (<b>c</b>) top-view of the printed textured [<a href="#B57-lubricants-13-00098" class="html-bibr">57</a>].</p>
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<p>Average specific wear rate: (<b>a</b>) nylon and SCFRN tested under dry and water lubricated conditions with a constant load of 20 N and (<b>b</b>) SCFRN tested under dry and water lubricated conditions with various loads [<a href="#B57-lubricants-13-00098" class="html-bibr">57</a>].</p>
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23 pages, 1565 KiB  
Review
Challenges and Advances in the Encapsulation of Bioactive Ingredients Using Whey Proteins
by Manuel Figueiredo, Zsuzsa Sárkány, Fernando Rocha and Pedro M. Martins
Foods 2025, 14(4), 691; https://doi.org/10.3390/foods14040691 - 17 Feb 2025
Viewed by 306
Abstract
Functional foods represent an emerging trend in the food industry. Fortifying foods with bioactive ingredients results in health benefits and reduces the risk of disease. Encapsulation techniques protect sensitive ingredients from degradation due to heat, light, moisture and other factors. Among encapsulating materials, [...] Read more.
Functional foods represent an emerging trend in the food industry. Fortifying foods with bioactive ingredients results in health benefits and reduces the risk of disease. Encapsulation techniques protect sensitive ingredients from degradation due to heat, light, moisture and other factors. Among encapsulating materials, milk whey proteins are particularly attractive due to their availability, GRAS status and remarkable ligand-binding ability. Whey protein was once considered a by-product in the dairy industry but is now seen as a promising resource given its natural role as a nutrient carrier. This work reviews the encapsulation systems that employ whey proteins in the food industry. The structural features of β-lactoglobulin (β-LG), the main protein constituent of milk whey, are presented in the context of its ligand-binding properties. Different types of encapsulation systems using whey proteins are discussed, focusing on the recent advances in stable formulations of bioactives using whey protein, alone or in hybrid systems. Whey proteins are a valuable asset capable of binding sensitive bioactive compounds such as vitamins, polyphenols and antioxidants and forming stable complexes that can be formulated as nanoparticles, nanofibrils, emulsions and other micro- and nanostructures. Developing scalable, solid and stable encapsulation systems is identified as a main challenge in the field. Full article
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Figure 1
<p>Retinol binds into the calyx, the main hydrophobic binding site of β-LG. (<b>A</b>) The overall crystal structure of monomeric β-LG variant B in complex with retinol (PDB entry 1GX8). The β-LG structure is represented as cartoon where the colors highlight different types of secondary structures. The main fluorophore TRP19 is indicated at the bottom of the calyx as yellow sticks. Retinol is denoted as green sticks. (<b>B</b>) Electrostatic surface representation of β-LG. The surfaces are colored according to electrostatic potential contoured at ±8 kT/eV (blue, positive; red, negative). Left and right panels are related by 180° rotation. Images of β-LG: retinol complexes were prepared with Pymol (The PyMOL Molecular Graphics System, Version 2.5.0 Schrödinger, LLC, New York, NY, USA).</p>
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<p>Surface representation of monomeric β-LG variant B in complex with retinol (PDB entry 1GX8). Retinol is denoted as green sticks. Images of β-LG: retinol complexes were prepared with Pymol (The PyMOL Molecular Graphics System, Version 2.5.0 Schrödinger, LLC).</p>
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<p>Advantages and limitations of different types of whey protein nanocarriers. Images adapted from [<a href="#B74-foods-14-00691" class="html-bibr">74</a>].</p>
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15 pages, 1673 KiB  
Article
Prenatal SARS-CoV-2 Infection Alters Human Milk-Derived Extracellular Vesicles
by Somchai Chutipongtanate, Supasek Kongsomros, Hatice Cetinkaya, Xiang Zhang, Damaris Kuhnell, Desirée Benefield, Wendy D. Haffey, Michael A. Wyder, Gaurav Kwatra, Shannon C. Conrey, Allison R. Burrell, Scott M. Langevin, Leyla Esfandiari, David S. Newburg, Kenneth D. Greis, Mary A. Staat and Ardythe L. Morrow
Cells 2025, 14(4), 284; https://doi.org/10.3390/cells14040284 - 15 Feb 2025
Viewed by 454
Abstract
Human milk-derived extracellular vesicles (HMEVs) are key components in breast milk, promoting infant health and development. Maternal conditions could affect HMEV cargo; however, the impact of SARS-CoV-2 infection on HMEVs remains unknown. This study investigated the influence of SARS-CoV-2 infection during pregnancy on [...] Read more.
Human milk-derived extracellular vesicles (HMEVs) are key components in breast milk, promoting infant health and development. Maternal conditions could affect HMEV cargo; however, the impact of SARS-CoV-2 infection on HMEVs remains unknown. This study investigated the influence of SARS-CoV-2 infection during pregnancy on postpartum HMEV molecules. The median duration from SARS-CoV-2 test positivity to milk collection was 3 months. After defatting and casein micelle disaggregation, HMEVs were isolated from milk samples of nine mothers with prenatal SARS-CoV-2 and six controls by sequential centrifugation, ultrafiltration, and qEV-size exclusion chromatography. The presence of HMEV was confirmed via transmission electron microscopy. Nanoparticle tracking analysis demonstrated particle diameters of <200 nm and yields of >1 × 1011 particles per mL of milk. Western immunoblots detected ALIX, CD9, and HSP70, supporting the presence of HMEVs in the isolates. Cargo from thousands of HMEVs were analyzed using a multi-omics approach, including proteomics and microRNA sequencing, and predicted that mothers with prenatal SARS-CoV-2 infection produced HMEVs with enhanced functionalities involving metabolic reprogramming, mucosal tissue development, and immunomodulation. Our findings suggest that SARS-CoV-2 infection during pregnancy boosts mucosal site-specific functions of HMEVs, potentially protecting infants against viral infections. Further prospective studies should be pursued to reevaluate the short- and long-term benefits of breastfeeding in the post-COVID era. Full article
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<p>HMEV isolation and characterization. (<b>a</b>) The optimized HMEV isolation workflow. (<b>b</b>) Nanoparticle tracking analysis (NTA). (<b>c</b>) Transmission electron microscope (TEM). Scale bars, 1000 nm (left panel) and 200 nm (right panel), respectively. (<b>d</b>) Western immunoblotting against three common EV markers: Alix, Hsp70, and CD9. (<b>e</b>) Proteomics with PCA clearly separated HMEVs from HM, dHM, and MVs. (<b>f</b>) Relative protein intensities of three common EV markers and HMEV-specific markers [<a href="#B35-cells-14-00284" class="html-bibr">35</a>,<a href="#B36-cells-14-00284" class="html-bibr">36</a>], including butyrophilin, lactadherin, and XDH. dHM, defatted HM; Hsp70, heat shock protein 70; MV, microvesicle; PCA, principal component analysis; XDH, xanthine dehydrogenase.</p>
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<p>HMEV proteomic analysis revealed prenatal SARS-CoV-2 infection affected HMEV proteins at 2 weeks of lactation period. (<b>a</b>) Heatmap with unsupervised clustering of 52 significant proteins. Color scale represents protein fold-change over the median protein expression after log2 transformation. (<b>b</b>) Volcano plot showing 35 relevant proteins at the thresholds of 2× fold change and <span class="html-italic">p</span>-value &lt; 0.05. (<b>c</b>) Functional enrichment analysis of 35 HMEV-relevant proteins against GO biological process, KEGG, and reactome. Adjusted <span class="html-italic">p</span>-value &lt; 0.05 determined functional significance.</p>
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<p>HMEV miRNA analysis in prenatal SARS-CoV-2 infection versus control groups. (<b>a</b>) PCA based on all 2588 identified miRNAs. A total of 232 miRNAs passed a cutoff of 100 counts. (<b>b</b>) Mean-variance trend plot for quality control. (<b>c</b>) Volcano plot shows no differentially expressed miRNAs between groups. (<b>d</b>) Boxplot showing the read counts of the 20 most abundant miRNAs in HMEVs.</p>
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38 pages, 10117 KiB  
Article
Wild-Grown Romanian Eupatorium cannabinum: Advancing Phyto-Nanocarriers via Maltodextrin Micro-Spray Encapsulation—Metabolite Profiling, Antioxidant, Antimicrobial, and Cytotoxicity Insights
by Gabriela Vlase, Adina-Elena Segneanu, Ludovic Everard Bejenaru, Ionela Amalia Bradu, Crina Sicoe, Titus Vlase, George Dan Mogoşanu, Gabriela Buema, Dumitru-Daniel Herea, Maria Viorica Ciocîlteu and Cornelia Bejenaru
Polymers 2025, 17(4), 482; https://doi.org/10.3390/polym17040482 - 12 Feb 2025
Viewed by 471
Abstract
In Romanian ethnopharmacology, Eupatorium cannabinum species is known for its remarkable biological activity. We present an advanced approach to encapsulation using maltodextrin matrices to enhance the stability and efficacy of phytoconstituents and nanoparticles. Two distinct carrier systems were developed: (i) a direct micro-spray [...] Read more.
In Romanian ethnopharmacology, Eupatorium cannabinum species is known for its remarkable biological activity. We present an advanced approach to encapsulation using maltodextrin matrices to enhance the stability and efficacy of phytoconstituents and nanoparticles. Two distinct carrier systems were developed: (i) a direct micro-spray encapsulation of E. cannabinum in maltodextrin to produce a maltodextrin-encapsulated carrier (MEC), and (ii) a two-step process involving the preparation of a new phytocarrier system based on gold nanoparticles (EC-AuNPs), followed by micro-spray encapsulation in maltodextrin to create the maltodextrin-encapsulated AuNPs system (MEC-AuNPs system). Comprehensive chemical profiling using GC–MS and ESI–QTOF–MS revealed 80 bioactive molecules, including terpenoids, alkaloids, flavonoids, and phytoecdysteroids. Morpho-structural (XRD, FTIR, Raman spectroscopy, SEM) and thermal analyses confirmed the successful integration of NPs within the matrices. EC-AuNPs and MEC-AuNPs exhibited superior antioxidant activity, significant antimicrobial efficacy against major bacterial pathogens (S. aureus, B. subtilis, B. cereus, P. aeruginosa, S. typhi, and E. coli), and enhanced cytotoxicity against MCF-7 and HT-29 cancer cell lines. This study highlights the potential of combining E. cannabinum with AuNPs and maltodextrin encapsulation to develop multifunctional therapeutic systems. The findings underscore the importance of phytoconstituent stabilization and nanotechnology in addressing global antimicrobial resistance and advancing innovative medical applications. Full article
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<p>Biomolecules classification bar chart of <span class="html-italic">E. cannabinum</span> sample.</p>
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<p>The volatile organic compound (VOC) odor profile of <span class="html-italic">E. cannabinum</span>.</p>
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<p>FTIR spectra of <span class="html-italic">E. cannabinum</span> sample, EC-AuNPs system, MEC carrier, and MEC-AuNPs system. EC-AuNPs, <span class="html-italic">E. cannabinum</span>–gold nanoparticles system; FTIR, Fourier-transform infrared; MEC, maltodextrin–<span class="html-italic">E. cannabinum</span>.</p>
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<p>Powder XRD patterns of <span class="html-italic">E. cannabinum</span> sample (<b>a</b>), EC-AuNPs system (<b>b</b>), and overlapping XRD patterns of <span class="html-italic">E. cannabinum</span> and EC-AuNPs system (<b>c</b>). XRD, X-ray diffraction.</p>
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<p>SEM image of <span class="html-italic">E. cannabinum</span> (<b>a</b>), EC-AuNPs system (<b>b</b>), MEC carrier system (<b>c</b>), and MEC-AuNPs carrier system (<b>d</b>). SEM, scanning electron microscopy.</p>
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<p>HR-TEM image of AuNPs. HR-TEM, high-resolution transmission electron microscopy.</p>
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<p>EDX analysis of the <span class="html-italic">E. cannabinum</span> (<b>a</b>) and EC-AuNPs system (<b>b</b>). EDX, energy dispersive X-ray.</p>
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<p>DLS patterns of <span class="html-italic">E. cannabinum</span> (<b>a</b>) and EC-AuNPs system (<b>b</b>). DLS, dynamic light scattering.</p>
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<p>PSD curves for 10 consecutive measurements (taken over a 2 min period) for MEC carrier (<b>a</b>) and MEC-AuNPs system (<b>b</b>). PSD, particles size distribution.</p>
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<p>Themoanalytical curves of <span class="html-italic">E. cannabinum</span> sample (<b>a</b>), EC-AuNPs system (<b>b</b>), MEC carrier (<b>c</b>), and MEC-AuNPs system (<b>d</b>). DTG, differential thermogravimetry; HF, heat flow; TG, thermogravimetry.</p>
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<p>Themoanalytical curves of <span class="html-italic">E. cannabinum</span> sample (<b>a</b>), EC-AuNPs system (<b>b</b>), MEC carrier (<b>c</b>), and MEC-AuNPs system (<b>d</b>). DTG, differential thermogravimetry; HF, heat flow; TG, thermogravimetry.</p>
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<p>Schematic representation of TPC (<b>a</b>), FRAP (<b>b</b>), and DPPH (<b>c</b>) assays results for <span class="html-italic">E. cannabinum</span>, EC-AuNPs system, MEC carrier, and MEC-AuNPs system. DPPH, 2,2-Diphenyl-1-picrylhydrazyl; FRAP, ferric reducing antioxidant power; TPC, total phenolic content.</p>
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<p>Viability of HT-29 and MCF-7 cells, 24 h after co-incubation with different concentrations of <span class="html-italic">E. cannabinum</span> sample, EC-AuNPs system, MEC carrier, and MEC-AuNPs system. Positive control wells contained untreated cells, MTT solution, and DMSO. Data are represented as mean ± SD (<span class="html-italic">n</span> = 3). DMSO, dimethyl sulfoxide; MTT, 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide; SD, standard deviation.</p>
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<p>In vitro cytotoxicity of <span class="html-italic">E. cannabinum</span>, AuNPs, the EC-AuNPs system, the MEC carrier, and the MEC-AuNPs system, as a function of concentration against MCF-7 and HT-29 cells (after 24 h). Data are represented as mean ± SD (<span class="html-italic">n</span> = 3).</p>
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21 pages, 5045 KiB  
Article
Comparative Toxicity of Micro, Nano, and Leachate Fractions of Three Rubber Materials to Freshwater Species: Zebrafish and Daphnia
by Miranda E. Jackson, Bryan J. Harper, Manuel Garcia-Jaramillo and Stacey L. Harper
Microplastics 2025, 4(1), 8; https://doi.org/10.3390/microplastics4010008 - 11 Feb 2025
Viewed by 609
Abstract
Rubber materials enter aquatic environments by stormwater runoff via sources such as playground mulch, athletic fields, and roadway surfaces. Tire rubbers are considered plastics as they comprise a substantial portion of synthetic polymers. Rubber particles are complex and variable depending on the type, [...] Read more.
Rubber materials enter aquatic environments by stormwater runoff via sources such as playground mulch, athletic fields, and roadway surfaces. Tire rubbers are considered plastics as they comprise a substantial portion of synthetic polymers. Rubber particles are complex and variable depending on the type, source, and age of rubber. In this study, zebrafish embryos and daphnids were exposed to nano-scale or micro-scale particles, or leachate from recycled rubber (RR), crumb rubber (CR), and cryo-milled tire tread (CMTT). Zebrafish embryos were evaluated for lethal and sub-lethal effects over a 120 h exposure, while daphnids were tested over a 48 h period. Nano-scale RR, CR, and CMTT particles elicited a hatch delay in zebrafish embryos with similar EC50 values (1.3 × 109–1.4 × 109 particles/mL). Micro-scale particles did not elicit any significant effects in developing zebrafish. Nano-scale particles of all rubber materials significantly increased hatch delay compared to leachate, suggesting an adverse nanoparticle effect unexplained by chemical leaching alone, indicating tire particle-specific effects. Daphnia RR micro- and nanoparticle exposures resulted in mortality, with LC50 values of 9.8 × 105 microparticles/mL and 5.0 × 108 nanoparticles/mL, respectively. Leachate exposures did not elicit significant Daphnia mortality. Sublethal micro- and nano-TP exposures significantly decreased microalgae ingestion by Daphnia after 24 h. The effects of tire-derived exposures observed pose a risk to aquatic organism survival at environmentally relevant concentrations. Full article
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<p>Experimental design describing tire particle size fraction filtration, particle and leachate characterization, and biological toxicity testing.</p>
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<p>SEM images of cryo-milled microparticle recycled rubber (RR), crumb rubber (CR), and pristine cryo-milled tire tread (CMTT).</p>
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<p>ICP-OES metals and limit of detection (LOD) concentrations in µg/L of RR, CR, and CMTT exposure leachate samples in fish water and Daphnia water with Euclidean clustering. Daphnia acute 48 h EC<sub>50</sub> toxicity information [<a href="#B53-microplastics-04-00008" class="html-bibr">53</a>] and broad aquatic toxicity information from the USEPA’s Hazard Module: <a href="https://hcd.rtpnc.epa.gov/#/hazard/report" target="_blank">https://hcd.rtpnc.epa.gov/#/hazard/report</a> accessed on 12 September 2024.</p>
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<p>Zebrafish hatch delay after 72 hpf concentration–response curves with 95% confidence intervals for (<b>A</b>) nano-scale tire rubber particles and (<b>B</b>) tire rubber leachate. Letters denote significance between materials based on a one-way ANOVA and Tukey post hoc analysis.</p>
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<p>Percentage leachate zebrafish hatch delay EC<sub>50</sub> values and standard error bars of nanoparticle (RR+, CR+, CMTT+) and leachate-only (RR, CR, CMTT) fractions. Asterisks denote significance between groups based on a two-way ANOVA.</p>
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<p>Daphnia 48 h mortality and immobility concentration-response curves with shaded areas representing the 95% confidence intervals for (<b>A</b>) nano-scale tire rubber particles and (<b>B</b>) micro-scale tire rubber particles. Letters denote significance between rubber types based on Fisher’s exact test.</p>
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<p><span class="html-italic">Daphnia</span> microalgal ingestion after 24 h of TP exposure with SD error bars: (<b>A</b>) micro-TP exposure; (<b>B</b>) nano-TP exposure. Asterisks denote significance from control based on a two-way ANOVA. <span class="html-italic">Daphnia</span> images from left to right include directly after 24 h RR exposure, 24 h unfed CMTT depuration, and 24 h fed CR depuration.</p>
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14 pages, 10364 KiB  
Article
SnO2-Based CMOS-Integrated Gas Sensor Optimized by Mono-, Bi-, and Trimetallic Nanoparticles
by Larissa Egger, Florentyna Sosada-Ludwikowska, Stephan Steinhauer, Vidyadhar Singh, Panagiotis Grammatikopoulos and Anton Köck
Chemosensors 2025, 13(2), 59; https://doi.org/10.3390/chemosensors13020059 - 8 Feb 2025
Viewed by 524
Abstract
Chemical sensors, relying on electrical conductance changes in a gas-sensitive material due to the surrounding gas, have the (dis-)advantage of reacting with multiple target gases and humidity. In this work, we report CMOS-integrated SnO2 thin film-based gas sensors, which are functionalized with [...] Read more.
Chemical sensors, relying on electrical conductance changes in a gas-sensitive material due to the surrounding gas, have the (dis-)advantage of reacting with multiple target gases and humidity. In this work, we report CMOS-integrated SnO2 thin film-based gas sensors, which are functionalized with mono-, bi-, and trimetallic nanoparticles (NPs) to optimize the sensor performance. The spray pyrolysis technology was used to deposit the metal oxide sensing layer on top of a CMOS-fabricated micro-hotplate (µhp), and magnetron sputtering inert-gas condensation was employed to functionalize the sensing layer with metallic NPs, Ag-, Pd-, and Ru-NPs, and all combinations thereof were used as catalysts to improve the sensor response to carbon monoxide and to suppress the cross-sensitivity toward humidity. The focus of this work is the detection of toxic carbon monoxide and a specific hydrocarbon mixture (HCmix) in a concentration range of 5–50 ppm at different temperatures and humidity levels. The use of CMOS chips ensures low-power, integrated sensors, ready to apply in cell phones, watches, etc., for air quality-monitoring purposes. Full article
(This article belongs to the Special Issue Advanced Chemical Sensors for Gas Detection)
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<p>(<b>a</b>) Micro-hotplate chip and (<b>b</b>) single µhp.</p>
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<p>High-resolution TEM investigation of (<b>a</b>) Ru NP, (<b>b</b>) Ag NP, and (<b>c</b>) Pd NP.</p>
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<p>TEM investigation of (<b>a</b>) Ru NP, (<b>b</b>) Ag NP, and (<b>c</b>) Pd NP and the size distribution of (<b>d</b>) Ru NP, (<b>e</b>) Ag NP, and (<b>f</b>) Pd NP.</p>
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<p>TEM investigation of (<b>a</b>) PdRu NP, (<b>b</b>) AgPd NP, and (<b>c</b>) AgRu NP and the size distribution of (<b>d</b>) PdRu NP, (<b>e</b>) AgPd NP, and (<b>f</b>) AgRu NP.</p>
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<p>TEM investigation of (<b>a</b>) AgPdRu NP and the size distribution of (<b>b</b>) AgPdRu NP.</p>
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<p>Resistance measurement of a bare SnO<sub>2</sub> sensor operated at 200 °C. The background gas has three different humidity levels: 50%, 25%, and 75%. At each humidity level, three CO gas pulses (duration 5 min) with 5 ppm, 25 ppm, and 50 ppm concentrations are introduced.</p>
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<p>Response of a bare SnO<sub>2</sub> sensor at 200 °C operation temperature to three CO test gas pulses with 5 ppm (black), 25 ppm (red), and 50 ppm concentration (blue) at humidity levels of 25%, 50%, and 75%.</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub> functionalized with monometallic Ag (red), Pd (blue), and Ru (green) NPs to different CO concentrations (5 ppm, 25 ppm, and 50 ppm) at different humidity levels ((<b>a</b>) 25%, (<b>b</b>) 50%, and (<b>c</b>) 75%).</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub>-functionalized bimetallic AgPd, RuPd, and AgRu NPs to different CO concentrations (5 ppm, 25 ppm, and 50 ppm) at different humidity levels ((<b>a</b>) 25%, (<b>b</b>) 50%, and (<b>c</b>) 75%).</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub> functionalized with Ru, AgRu, and PdAgRu NPs to different CO concentrations (5 ppm, 25 ppm, and 50 ppm) at different humidity levels ((<b>a</b>) 25%, (<b>b</b>) 50%, and (<b>c</b>) 75%).</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub> functionalized with (<b>a</b>) monometallic, (<b>b</b>) bimetallic, and (<b>c</b>) trimetallic NP-functionalized sensors to 50 ppm CO concentrations at different humidity levels (25%, 50%, and 75%).</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub> functionalized with monometallic NP to different HC<sub>mix</sub> concentrations (5 ppm, 25 ppm, and 50 ppm) at different humidity levels ((<b>a</b>) 25%, (<b>b</b>) 50%, and (<b>c</b>) 75%).</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub> functionalized with bimetallic NPs to different HC<sub>mix</sub> concentrations (5 ppm, 25 ppm, and 50 ppm) at different humidity levels ((<b>a</b>) 25%, (<b>b</b>) 50%, and (<b>c</b>) 75%).</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub> functionalized with Ru, AgRu, and PdAgRu NPs to different HC<sub>mix</sub> concentrations (5 ppm, 25 ppm, and 50 ppm) at different humidity levels ((<b>a</b>) 25%, (<b>b</b>) 50%, and (<b>c</b>) 75%).</p>
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<p>Response of a bare SnO<sub>2</sub> sensor (black) and SnO<sub>2</sub> functionalized with (<b>a</b>) monometallic, (<b>b</b>) bimetallic, and (<b>c</b>) trimetallic NPs to 50 ppm HC<sub>mix</sub> concentrations at different humidity levels (25%, 50%, and 75%).</p>
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<p>Relative change in the responses of the functionalized sensors compared to the bare SnO<sub>2</sub> for (<b>a</b>–<b>c</b>) 50 ppm CO and (<b>d</b>–<b>f</b>) 50 ppm HC<sub>mix</sub> concentrations at different humidity levels ((<b>a</b>,<b>d</b>) 25%, (<b>b</b>,<b>e</b>) 50%, and (<b>c</b>,<b>f</b>) 75%).</p>
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16 pages, 2069 KiB  
Review
Retinopathy of Prematurity and MicroRNAs
by Giuseppe Maria Albanese, Giacomo Visioli, Ludovico Alisi, Marta Armentano, Francesca Giovannetti, Luca Lucchino, Marco Marenco, Paola Pontecorvi and Magda Gharbiya
Biomedicines 2025, 13(2), 400; https://doi.org/10.3390/biomedicines13020400 - 7 Feb 2025
Viewed by 491
Abstract
Retinopathy of Prematurity (ROP), a leading cause of blindness in preterm infants, arises from dysregulated angiogenesis and inflammation. Without timely intervention, ROP can progress to severe outcomes, including dense fibrovascular plaques and retinal detachment. MicroRNAs (miRNAs) regulate key pathways such as hypoxia response, [...] Read more.
Retinopathy of Prematurity (ROP), a leading cause of blindness in preterm infants, arises from dysregulated angiogenesis and inflammation. Without timely intervention, ROP can progress to severe outcomes, including dense fibrovascular plaques and retinal detachment. MicroRNAs (miRNAs) regulate key pathways such as hypoxia response, VEGF signaling, and vascular remodeling. Studies have identified miRNAs (e.g., miR-210, miR-146a, and miR-21) as potential biomarkers and therapeutic targets. Preclinical evidence supports miRNA-based therapies (e.g., miR-18a-5p and miR-181a), targeting HIF-1α and VEGFA to mitigate neovascularization, with nanoparticle delivery systems enhancing stability and specificity. These strategies, combined with anti-VEGF agents, show significant potential for improving ROP management. While promising, miRNA therapies require validation in clinical trials to ensure safety and efficacy. This review discusses the role of miRNAs in ROP, highlighting their relevance as diagnostic and therapeutic tools. Full article
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<p>Schematic representation of the development of retinal neovessels in Retinopathy of Prematurity. During the intrauterine stage, vessels develop normally, but in the case of preterm birth, vascularization remains incomplete. Compared to the intrauterine stage, from birth to the 31st gestational week, the retina is exposed to relative hyperoxia, leading to the inhibition of VEGF and IGF-1. However, between 31 and 34 weeks, as the metabolic demand of the retina increases, it becomes exposed to relative hypoxia, triggering excessive VEGF production and the formation of neovessels [<a href="#B16-biomedicines-13-00400" class="html-bibr">16</a>].</p>
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<p>Schematic representation of the retinal zones used to classify the location of Retinopathy of Prematurity (ROP) according to the International Classification of Retinopathy of Prematurity (ICROP). The zones are defined as follows: Zone I represents the posterior retina within a circle centered on the optic nerve; Zone II extends from the edge of Zone I to the nasal ora serrata; and Zone III forms the outermost crescent of the retina.</p>
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<p>Retinography of a premature infant with aggressive Retinopathy of Prematurity (A-ROP), demonstrating marked vascular tortuosity characteristic of plus disease, vascular loops (white arrows), and flat extraretinal vascularization (white arrowheads). A-ROP, as defined in the third edition of the International Classification of ROP (ICROP3), includes rapidly progressing forms such as aggressive posterior ROP (AP-ROP), which can bypass the typical staged progression of the disease.</p>
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<p>Schematic representation of the roles of key miRNAs in Retinopathy of Prematurity (ROP).</p>
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26 pages, 2538 KiB  
Review
Non-Invasive Delivery of CRISPR/Cas9 Ribonucleoproteins (Cas9 RNPs) into Cells via Nanoparticles for Membrane Transport
by Toshihiko Tashima
Pharmaceutics 2025, 17(2), 201; https://doi.org/10.3390/pharmaceutics17020201 - 6 Feb 2025
Viewed by 587
Abstract
The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system is a promising biotechnology tool for genome editing. However, in living organisms, several pharmacokinetic challenges arise, including off-target side effects due to incorrect distribution, low bioavailability caused by membrane impermeability, and instability [...] Read more.
The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system is a promising biotechnology tool for genome editing. However, in living organisms, several pharmacokinetic challenges arise, including off-target side effects due to incorrect distribution, low bioavailability caused by membrane impermeability, and instability resulting from enzymatic degradation. Therefore, innovative delivery strategies must be developed to address these issues. Modified nanoparticles offer a potential solution for the non-invasive delivery of CRISPR/Cas9 ribonucleoproteins (Cas9 RNPs). Cas9 RNPs encapsulated in nanoparticles are protected from enzymatic degradation, similar to how microRNAs are shielded within exosomes. It is well-established that certain materials, including proteins, are expressed selectively in specific cell types. For example, the α-7 nicotinic receptor is expressed in endothelial and neuronal cells, while the αvβ3 integrin is expressed in cancer cells. These endogenous materials can facilitate receptor-mediated endocytosis or transcytosis. Nanoparticles encapsulating Cas9 RNPs and coated with ligands targeting such receptors may be internalized through receptor-mediated mechanisms. Once internalized, Cas9 RNPs could perform the desired gene editing in the nucleus after escaping the endosome through mechanisms such as the proton sponge effect or membrane fusion. In this review, I discuss the potential and advantages of delivering Cas9 RNP-encapsulated nanoparticles coated with ligands through receptor-mediated endocytosis or transcytosis. Full article
(This article belongs to the Special Issue Nanoparticle-Mediated Targeted Drug Delivery Systems)
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<p>The interference mechanism of the type II CRISPR-Cas system involves the cleavage of target double-stranded DNA, originating from bacteria, at two endonuclease domains on the Cas9 protein: the HNH domain (named for its characteristic histidine and asparagine residues) and the RuvC domain (named after an <span class="html-italic">Escherichia coli</span> protein involved in DNA repair). CRISPR RNA (crRNA) and trans-activating CRISPR RNA (tracrRNA) recognize the target gene based on a specific protospacer adjacent motif (PAM).</p>
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<p>Single-guide RNA (sgRNA) is an engineered RNA molecule formed by combining CRISPR RNA (crRNA) and trans-activating CRISPR RNA (tracrRNA). PAM refers to the protospacer adjacent motif. The HNH domain is an endonuclease domain named for its characteristic histidine and asparagine residues, while the RuvC domain is an endonuclease domain named after a protein in <span class="html-italic">Escherichia coli</span> that plays a role in DNA repair.</p>
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<p>The pathway of NTLA-2001 involves lipid nanoparticles encapsulating Cas9 messenger RNA (mRNA) and single-guide RNA (sgRNA) targeting transthyretin (TTR), coated with apolipoprotein E (APOE) protein. APOE binds to low-density lipoprotein receptor (LDLR) to facilitate endocytosis. Endosomal escape is likely achieved through the breakdown of the lipid nanoparticles and the disruption of the endosomal membrane.</p>
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<p>The structures of lipid nanoparticle components include 2,3-dioleoyl-glycero-1-phosphocholine (DOPC), 1,2-dimyristoyl-rac-glycero-3-methoxypolyethyleneglycol-2000 (DMG-PEG), cholesterol, 1,10-[[2-[4-[2-[2-[bis(2-hydroxydodecyl)amino]ethylamino]ethyl]piperazin-1-yl]ethyl]azanediyl]bis(dodecan-2-ol) (C12-200), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP).</p>
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<p>The outline of a lipid nanoparticle (LNP)-based Cas9 RNP delivery system uses optimally designed single-stranded oligonucleotides (ssODNs). The mechanism of endosomal escape is unknown; however, it is likely due to membrane fusion, although this remains uncertain.</p>
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<p>The structures of lipid nanoparticle components such as 5A2-SC8, 1,2-dioleyloxy-3-dimethylaminopropane (DODMA), and 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).</p>
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<p>The pathway of LNPs encapsulating Cas9 ribonucleoprotein (Cas9 RNP), surrounded by cationic lipids such as 5A2-SC8 and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) (depicted by a red circle), is described. Cas9 RNP is a complex of Cas9 protein and single-guide RNA (sgRNA). The LNPs are internalized into cells via lipid raft-mediated endocytosis. Although the mechanisms of endosomal escape remain unknown, DOTAP, as a permanent cationic lipid, may disrupt the membranes of both LNPs and endosomes.</p>
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<p>The structure of the nanoassembly is composed of Cas9 ribonucleoproteins (RNPs) with an oligo (20) Glu tag (Cas9E20) and a gold nanoparticle, linked through electrostatic connections between Glu (purple) and Arg (yellow).</p>
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<p>The pathway of nanoparticles, composed of the Cas9 ribonucleoprotein (RNP) formed by Cas9 and single-guide RNA (sgRNA), and the single-stranded oligonucleotide (ssODN) self-assembled with mPEG-PC7A through hydrophobic interactions, involves the release of the Cas9 RNP and ssODN complex into the cytosol via endocytosis. This process is followed by self-disassembly due to acidification in the endosomes and endosomal escape based on the proton sponge effect. As indicated in yellow, proton acceptance by amines in the endosomes triggers an influx of chloride ions and water, leading to osmotic rupture of the endosomal/lysosomal membrane.</p>
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<p>The pathway of exosome<sup>RNP</sup> nanocomplex internalization involves endocytosis and endosomal escape of the Cas9 ribonucleoprotein (RNP), composed of Cas9 and single-guide RNA (sgRNA), with these components acting as their cargos via membrane fusion.</p>
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<p>The interaction between leukemia stem cells and injected mesenchymal stem cell membrane-coated nanofibrils (MSCM-NFs) loaded with LNPs encapsulating Cas9 RNP targeting the critical genes interleukin-1 receptor accessory protein (IL1RAP) and CXCL12α was studied. CXCL12α, a ligand for C-X-C chemokine receptor 4 (CXCR4), recruited leukemia stem cells. As a result, vascular cell adhesion molecule 1 (VCAM-1) derived from the mesenchymal stem cell membrane bound integrin α4 (VLA-4) on the surface of leukemia stem cells. Endocytosed LNPs encapsulating Cas9 RNP induced efficient gene editing in leukemia stem cells following endosomal escape.</p>
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9 pages, 1300 KiB  
Article
Production of Uniform Droplets and Lipid Nanoparticles Using Perfluoropolyether-Based Microfluidic Devices
by Mincheol Cho, Eun Seo Kim, Tae-Kyung Ryu, Inseong Choi and Sung-Wook Choi
Micromachines 2025, 16(2), 179; https://doi.org/10.3390/mi16020179 - 31 Jan 2025
Viewed by 743
Abstract
Microfluidic devices are greatly affected by the materials used. The materials used in previous studies had problems in various aspects, such as processing, adsorption, and price. This study will investigate the materials needed to overcome such problems. Various microfluidic devices based on the [...] Read more.
Microfluidic devices are greatly affected by the materials used. The materials used in previous studies had problems in various aspects, such as processing, adsorption, and price. This study will investigate the materials needed to overcome such problems. Various microfluidic devices based on the perfluorinated compound perfluoropolyether (PFPE) were fabricated and mixed with hydrophilic and amphiphilic monomers, including poly(ethylene glycol) diacrylate, polyethylene glycol monomethacrylate, poly(ethylene glycol) methyl ether methacrylate, acrylic acid, and 2-hydroxyethyl methacrylate. A PFPE-based sheet with a repeating structure of hydrophobic and hydrophilic groups was fabricated. Thus, the hydrophilicity of highly hydrophobic PFPE was enhanced. The fluidic channel was engraved on a PFPE-based sheet using laser cutting and a fabricated microfluidic device. The channels of microfluidic devices are micro-scale (100 µm~300 µm). The lipid nanoparticles and droplets generated through the microfluidic device demonstrated uniform particles continuously. Full article
(This article belongs to the Special Issue Microfluidic Nanoparticle Synthesis)
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<p>Schematic illustration of (<b>A</b>) fabrication, (<b>B</b>) emulsion, and (<b>C</b>) the production of LNPs in a microfluidic device.</p>
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<p>(<b>A</b>) Water contact angles, (<b>B</b>) swelling ratio, and (<b>C</b>) photographs of PFPE-based slabs with different contents of PFPE-based materials (<span class="html-italic">n</span> = 5).</p>
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<p>(<b>A</b>) Snapshot of PFPE-based slab channel, where fabricating LNPs were captured by high-speed camera and slab design for fabricating LNPs. (<b>B</b>) Z-average size and PDI of LNPs obtained by different PFPE-based material slabs.</p>
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<p>(<b>A</b>,<b>B</b>) Average diameter and CV value of the emulsions at different flow rates of the continuous phase, which was fabricated by different PFPE-based slabs, and the frequency graph of the emulsion diameter formed at TFR (1.7 mL/h) and FRR (2:15) flow rate conditions.</p>
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19 pages, 3219 KiB  
Article
Impact of UV-Irradiated Mesoporous Titania Nanoparticles (mTiNPs) on Key Onco- and Tumor Suppressor microRNAs of PC3 Prostate Cancer Cells
by Andrea Méndez-García, Luis Alberto Bravo-Vázquez, Padmavati Sahare and Sujay Paul
Genes 2025, 16(2), 148; https://doi.org/10.3390/genes16020148 - 25 Jan 2025
Viewed by 254
Abstract
Background: Mesoporous titanium dioxide nanoparticles (mTiNPs) are known for their chemical stability, non-toxicity, antimicrobial and anticancer effects, as well as for their photocatalytic properties. When this material is subjected to UV radiation, its electronic structure shifts, and during that process, reactive oxygen species [...] Read more.
Background: Mesoporous titanium dioxide nanoparticles (mTiNPs) are known for their chemical stability, non-toxicity, antimicrobial and anticancer effects, as well as for their photocatalytic properties. When this material is subjected to UV radiation, its electronic structure shifts, and during that process, reactive oxygen species are generated, which in turn exert apoptotic events on the cancer cells. Objectives: We evaluated the cytotoxic effects of UV-irradiated mTiNPs on prostate cancer (PCa) cell line PC3 with the aim of demonstrating that the interaction between UV-light and mTiNPs positively impacts the nanomaterial’s cytotoxic efficiency. Moreover, we assessed the differential expression of key oncomiRs and tumor suppressor (TS) miRNAs, as well as their associated target genes, in cells undergoing this treatment. Methods: PBS-suspended mTiNPs exposed to 290 nm UV light were added at different concentrations to PC3 cells. Cell viability was determined after 24 h with a crystal violet assay. Then, the obtained IC50 concentration of UV-nanomaterial was applied to a new PC3 cell culture, and the expression of a set of miRNAs and selected target genes was evaluated via qRT-PCR. Results: The cells exposed to photo-activated mTiNPs required 4.38 times less concentration of the nanomaterial than the group exposed to non-irradiated mTiNPs to achieve the half-maximal inhibition, demonstrating an improved cytotoxic performance of the UV-irradiated mTiNPs. Moreover, the expression of miR-18a-5p, miR-21-5p, and miR-221-5p was downregulated after the application of UV-mTiNPs, while TS miR-200a-5p and miR-200b-5p displayed an upregulated expression. Among the miRNA target genes, PTEN was found to be upregulated after the treatment, while BCL-2 and TP53 were underexpressed. Conclusions: Our cytotoxic outcomes coincided with previous reports performed in other cancer cell lines, strongly suggesting UV-irradiated mTiNPs as a promising nano-therapeutic approach against PCa. On the other hand, to the best of our knowledge, this is the first report exploring the impact of UV-irradiated mTiNPs on key onco- and TS microRNAs in PCa cells. Full article
(This article belongs to the Section Human Genomics and Genetic Diseases)
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<p>The methodology was followed during the application of mTiNP treatments to PC3 cells and in the subsequent evaluation of miRNA/target gene expression. (<b>A</b>) UV-irradiated mTiNPs and (<b>B</b>) non-UV-irradiated mTiNPs (created with a licensed version of Biorender).</p>
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<p>Results of the characterization of the TiNPs: (<b>a</b>,<b>b</b>) SEM images of synthesized mTiNPs; (<b>c</b>) particle size distribution of mTiNPs; (<b>d</b>) nitrogen adsorption/desorption isotherms; (<b>e</b>) pore size distribution of the mTiNPs; (<b>f</b>) FTIR analysis of dry mTiNPs.</p>
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<p>Microscopic images of PC3 cells at 40× magnification. (<b>A</b>) Negative Control; (<b>B</b>) Positive Control (without serum); (<b>C</b>) 10 µg/mL non-UV; (<b>D</b>) 10 µg/mL UV; (<b>E</b>) 120 µg/mL non-UV; (<b>F</b>) 120 µg/mL UV.</p>
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<p>Viable PC3 cells (%) after 24 h exposure to different concentrations of UV irradiated mTiNPs (orange) and mTiNPs without photocatalytic treatment (purple) (<b>A</b>). The viability of HEK 293 to different concentrations of UV-irradiated mTiNPs is also shown in (<b>B</b>).</p>
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<p>miRNA expression profile of PC3 cells treated with UV-irradiated mTiNPs. The analysis was performed via qPCR on total RNA samples isolated from PC3 cells that had been previously subjected to UV-irradiated mTiNPs (48.96 µg mL<sup>−1</sup>) for 24 h. The U6 was used as the endogenous control for normalization. Each bar graph represents the mean value of the relative fold changes ± the standard error of the biological replicates (* <span class="html-italic">p</span>-value &lt; 0.05).</p>
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<p>qPCR-based target gene expression in PC3 cells treated with UV-irradiated mTiNPs. The bars represent the mean value of the relative fold changes ± the standard error of the biological replicates (* <span class="html-italic">p</span>-value &lt; 0.05).</p>
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<p>Schematic illustration of the chemical reactions mTiNPs undergo when excited with UV light and the change in miRNA and associated gene expression when in contact with PCa cells. mTiNPs, immersed in an aqueous solution containing water and oxygen molecules, are irradiated with UV light, causing the electrons and holes to transition to the conduction band, generating an electronic shift that allows the generation of reactive oxygen species within PCa cells, causing a differential expression of the indicated miRNAs and associated target genes (created with a licensed version of Biorender).</p>
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32 pages, 10992 KiB  
Article
Small Extracellular Vesicles from Breast Cancer Cells Induce Cardiotoxicity
by Jhon Jairo Osorio-Méndez, Luis Alberto Gómez-Grosso, Gladis Montoya-Ortiz, Susana Novoa-Herrán and Yohana Domínguez-Romero
Int. J. Mol. Sci. 2025, 26(3), 945; https://doi.org/10.3390/ijms26030945 - 23 Jan 2025
Viewed by 304
Abstract
Cardiovascular diseases and cancer are leading global causes of morbidity and mortality, necessitating advances in diagnosis and treatment. Doxorubicin (Doxo), a potent chemotherapy drug, causes long-term heart damage due to cardiotoxicity. Small extracellular vesicles (sEVs) carry bioactive molecules—such as proteins, lipids, and nucleic [...] Read more.
Cardiovascular diseases and cancer are leading global causes of morbidity and mortality, necessitating advances in diagnosis and treatment. Doxorubicin (Doxo), a potent chemotherapy drug, causes long-term heart damage due to cardiotoxicity. Small extracellular vesicles (sEVs) carry bioactive molecules—such as proteins, lipids, and nucleic acids—that can modulate gene expression and signaling pathways in recipient cells, including cardiomyocytes. Through the delivery of cytokines, microRNAs, and growth factors, sEVs can influence cell survival, which plays a critical role in the development of cardiotoxicity. This study investigates the role of sEVs derived from breast cancer cells treated or not with Doxo and their potential to induce cardiomyocyte damage, thereby contributing to cardiotoxicity. We isolated sEVs from MCF-7 cells treated or not to Doxo using ultracentrifugation and characterized them through Nanoparticle Tracking Analysis (NTA), Scanning Electron Microscopy (SEM), and Western Blotting (WB) for the markers CD63, CD81, and TSG101. We analyzed cytokine profiles using a Multiplex Assay and Cytokine Membrane Array. We exposed Guinea pig cardiomyocytes to different concentrations of sEVs. We assessed their viability (MTT assay), shortening, reactive oxygen species (ROS–DHE dye) production, mitochondrial membrane potential (JC-1 dye), and calcium dynamics (FLUO-4 dye). We performed statistical analyses, including t-tests, ANOVA, Cohen’s d, and η2 to validate the robustness of the results. Treatment of MCF-7 cells with 0.01 μM Doxorubicin resulted in increased sEVs production, particularly after 48 h of exposure (~1.79 × 108 ± 2.77 × 107 vs. ~5.1 × 107 ± 1.28 × 107 particles/mL, n = 3, p = 0.0019). These sEVs exhibited protein profiles in the 130–25 kDa range and 93–123 nm sizes. They carried cytokines including TNF-α, IL-1β, IL-4, IFN-γ, and IL-10. Exposure of cardiomyocytes to sEVs (0.025 μg/mL to 2.5 μg/mL) from both Doxo-treated and untreated cells significantly reduced cardiomyocyte viability, shortened cell length by up to 20%, increased ROS production, and disrupted calcium homeostasis and mitochondrial membrane potential, indicating severe cellular stress and cardiotoxicity. These findings suggest that Doxo enhances sEVs production from breast cancer cells, which plays a key role in cardiotoxicity through their cytokine cargo. The study highlights the potential of these sEVs as biomarkers for early cardiotoxicity detection and as therapeutic targets to mitigate cardiovascular risks in chemotherapy patients. Future research should focus on understanding the mechanisms by which Doxorubicin-induced sEVs contribute to cardiotoxicity and exploring their diagnostic and therapeutic potential to improve patient safety and outcomes in cancer therapy. Full article
(This article belongs to the Special Issue Exosomes and Non-Coding RNA Research in Health and Disease)
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<p>Determination of Doxo concentration to obtain conditioned media and subsequent isolation of sEVs. (<b>A</b>) Representative photographs of MCF-7 cells unexposed (Control) and exposed to Doxo (0.01 and 0.07 µM) at 10<span class="html-italic">X</span> and 20<span class="html-italic">X</span> magnifications at 48 h. (<b>B</b>) Evaluation of MCF-7 cell viability at 24-, 48-, and 72-h post-treatment (hpt) with different concentrations of Doxo (0, 0.01, 0.03, 0.05, and 0.07 µM). (<b>C</b>) Assessment of the effect of different concentrations of Doxo on the number of MCF-7 cells after 24, 48, and 72 hpt with different concentrations of Doxo. The viability estimations represent three replicates from three independent biological samples (three different cell cultures and treatments). We performed significance analysis of the assays using ANOVA with significance levels denoted as * for <span class="html-italic">p</span> &lt; 0.05, ** for <span class="html-italic">p</span> &lt; 0.01, and *** for <span class="html-italic">p</span> &lt; 0.001. These results suggest that Doxo’s concentration of 0.001 µM is useful for obtaining a conditioned medium to isolate sEVs.</p>
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<p>Physical and biochemical characterization of the sEVs population isolated from MCF-7 conditioned media. (<b>A</b>,<b>B</b>) Results of nanoparticle tracking analysis of sEVs isolated at 100,000× <span class="html-italic">g</span> from supernatants of MCF-7 cells treated or not with 0.01 µM Doxo for 48 h. The graph shows the mean (black line) and standard error of the mean from the five analyses (in red), along with the average concentration/size of FTLA per experiment. (<b>C</b>) SEM analysis showed a spheroid shape with a size of less than 200 nm (scale bar = 500 nm). The arrows show the sEVs. (<b>D</b>) Particle concentration at the three conditioning times (12, 24, and 48 h) by NTA analysis. (<b>E</b>) Particle size at each conditioning time (<b>F</b>) Quantification of protein content in intact sEV and lysed with 2% SDS and cycles of heating–cooling with liquid nitrogen. We performed significance analysis of the assays using ANOVA with significance levels denoted as * for <span class="html-italic">p</span> &lt; 0.05 and **** for <span class="html-italic">p</span> &lt; 0.0001. (<b>G</b>) Western blot analysis of sEVs markers (CD81, CD63, and TSG101) and control marker of exosomal origin (CYT C). The nanoparticle tracking analysis of isolated sEVs is representative of two different measurements. We evaluated the total cell extract from MCF-7 cells and the sEVs fraction from MCF-7 cells treated with DMSO (vehicle control) and Doxo for 12 h. Findings demonstrate successful isolation of sEVs with consistent size, shape, and protein markers across conditions, ensuring biological relevance.</p>
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<p>Cytokine profiling of 42 targets in sEVs from Doxo-treated and untreated MCF-7 cells. (<b>A</b>) Membrane cytokine arrays showing internal positive controls and highlighted cytokines (green boxes) with significant differential expression. (<b>B</b>) Mean pixel density analysis of differentially expressed cytokines. Data represent two independent experiments with biological duplicates (<span class="html-italic">n</span> = 2). Significance levels: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and **** <span class="html-italic">p</span> &lt; 0.0001. These results suggest that Doxo enhances the pro-inflammatory cytokine content of sEVs, potentially contributing to cardiotoxicity.</p>
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<p>Concentration and distribution profiles of six cytokines (IFN-γ, IL-1β, IL-4, IL-6, IL-10, TNF-α) across soluble, lysed, and unlysed sEV fractions from Doxo-treated and untreated (Ctrl) MCF-7 cells. Statistical analysis via ANOVA; significance levels: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and **** <span class="html-italic">p</span> &lt; 0.0001. Data include three technical replicates from three biological replicates per treatment condition (<span class="html-italic">n</span> = 3). These data highlight the role of cytokine compartmentalization in modulating sEVs-mediated effects on recipient cells.</p>
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<p>MTT reductase activity in isolated Guinea pig cardiomyocytes treated with different concentrations of vesicular proteins. This figure shows the corrected mean of formazan produced by cardiomyocytes incubated for 12 h with extracellular vesicles (sEVs) (100,000× <span class="html-italic">g</span>) derived from MCF-7 cells treated with 0.01 µM Doxo or the corresponding DMSO vehicle control. We evaluated the biological effects of sEVs using three different concentrations of vesicular protein: 0.025, 0.25, and 2.5 µg/mL, all dispersed in DMEM medium. The study exposed the control group of cardiomyocytes to DMEM medium only. (<b>A</b>) Formazan production results for cardiomyocytes treated with sEVs from control (0.25 µg/mL) and Doxo-treated (0.25 µg/mL) MCF-7 cells. (<b>B</b>) The figure shows the formazan production kinetics for each experimental group. (<b>C</b>) Statistical analysis was performed at 0, 50, and 70 min after MTT addition using a two-tailed unpaired Student’s <span class="html-italic">t</span>-test (<span class="html-italic">n</span> = 6). Significance levels were indicated as * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001. Results indicate that sEVs from Doxo-treated cells significantly impair cardiomyocyte metabolic activity compared to control sEVs.</p>
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<p>Effect of different concentrations of sEVs on the length of isolated Guinea pig cardiomyocytes. (<b>A</b>) A representative image shows cardiomyocyte shortening and loss of rod-like shape after treatment with 0.025 µg/mL concentration of sEVs from media conditioned for 12, 24, and 48 h. (<b>B</b>) Percentage of cardiomyocyte shortening treatment with a 0.025 µg/mL concentration of sEVs of three different conditioning times (12, 24, and 48 hc). Results represent three independent experiments using cardiomyocytes isolated from three different preparations and treated with sEVs (<span class="html-italic">n</span> = 15 per experiment). We performed statistical analysis using ANOVA for shortening after 48 h of treatment. Significance levels are indicated as * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and **** <span class="html-italic">p</span> &lt; 0.0001. Findings demonstrate that sEVs induce significant cardiomyocyte shortening, correlating with cellular stress and reduced viability.</p>
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<p>Effect of sEVs from three different conditioning times (12, 24, and 48 h of conditioning (hc)) on ROS (<b>A</b>,<b>B</b>), intracellular calcium (<b>C</b>,<b>D</b>), and mitochondrial membrane potential (<b>E</b>,<b>F</b>) in isolated Guinea pig cardiomyocytes. (<b>A</b>) Representative fluorescence microscopy images of cardiomyocytes in a bright field and fluorescence at approximately 585 to 590 nm. (<b>B</b>) Assessment of ROS production in isolated cardiomyocytes treated with 0.025 μg/mL of sEVs for 24 h. (<b>C</b>) Representative fluorescence microscopy images showing cardiomyocytes in a bright field and fluorescence at 514 to 529 nm. (<b>D</b>) Measurement of intracellular calcium levels in cardiomyocytes treated with 0.025 μg/mL of sEVs for 24 h. (<b>E</b>) Representative fluorescence microscopy images of cardiomyocytes in a bright field, with fluorescence at 514 to 529 nm and 585 to 590 nm. (<b>F</b>) Evaluation of mitochondrial membrane potential in cardiomyocytes treated with 0.025 μg/mL of sEVs for 24 h. ANOVA statistical analysis was performed on the mean fluorescence values for 15 cardiomyocytes per group (<span class="html-italic">n</span> = 15). The analysis used three measurements from three different isolations and treatments. Significance levels are indicated as * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001. We used the Shapiro–Wilk and Kolmogorov–Smirnov tests to assess normal data distribution and applied Bartlett’s test to assess equality of variance. ANOVA or ANOVA/Brown–Forsythe followed Bartlett’s test. Tukey’s or Tamhane’s T2 test was used for multiple comparisons. Effect sizes were determined using eta squared (η<sup>2</sup>) and Cohen’s d tests. These results underline the role of sEVs in inducing cardiomyocyte dysfunction through oxidative stress, calcium imbalance, and mitochondrial depolarization.</p>
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14 pages, 4687 KiB  
Article
Core–Shell Composite GaP Nanoparticles with Efficient Electroluminescent Properties
by Duo Chen, Ruiyuan Bi, Lifeng Xun, Xiaoyan Li, Qingyu Hai, Yao Qi and Xiaopeng Zhao
Materials 2025, 18(3), 487; https://doi.org/10.3390/ma18030487 - 22 Jan 2025
Viewed by 347
Abstract
Gallium-based light-emitting diodes (LEDs), including AlGaInP and GaN, have become the most widely used light-emitting devices in modern scientific research and practical applications. However, structures like carrier injection layers, active layers, and quantum well layers ensure the high luminescence efficiency of LEDs but [...] Read more.
Gallium-based light-emitting diodes (LEDs), including AlGaInP and GaN, have become the most widely used light-emitting devices in modern scientific research and practical applications. However, structures like carrier injection layers, active layers, and quantum well layers ensure the high luminescence efficiency of LEDs but also limit their applications at the micro- and nanoscale. Although the next generation of micrometer-scale light-emitting diodes (Micro-LEDs) has alleviated these issues to some extent, challenges such as edge effects and etching damage caused by size reduction lead to lower luminous efficiency and shorter lifetimes. Inspired by LED structure, this study designed and synthesized core–shell composite GaP:Zn/GaP/GaInP and GaP:Te/GaP nanoparticles using a thermal injection method. After high-temperature annealing, these composite materials demonstrated efficient electroluminescent performance under electric field excitation through band-edge transitions and the ZnGa-OP recombination mechanism. Experimental results show that the GaP:Zn/GaP/GaInP-GaP:Te/GaP composite samples with doping concentrations of 15%Zn-8%Te, a core–shell precursor ratio of 1:1:1, and reaction times of 1 h:20 min:20 min exhibit the best electron–hole injection efficiency and bound-recombination efficiency. Under excitation by an external electric field, they demonstrated optimal electroluminescence performance, with a relative luminous intensity of 11,109.21 at 600 nm, approximately 15 times higher than that of the initial condition samples. In addition, this study systematically investigated the structure, morphology, and elemental composition of the composite materials using various characterization techniques, including X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS). These GaP-doped nanoparticles with a core–shell composite structure, inspired by LED design, exhibited outstanding electroluminescent performance, providing new insights into the development of novel micro- and nanoscale electroluminescent materials. Full article
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<p>XRD patterns of GaP:15%Zn/GaP/GaInP and GaP:8%Te/GaP samples.</p>
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<p>TEM images and particle size distribution of GaP nanoparticles (The red circle highlights a single GaP nanoparticle). (<b>a</b>,<b>b</b>) GaP:Zn/GaP/GaInP. (<b>c</b>,<b>d</b>) GaP:Te/GaP. (<b>e</b>,<b>f</b>) Pure GaP core.</p>
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<p>SEM images and EDS spectra of GaP:Zn/GaP/GaInP and GaP:Te/GaP after mixing, annealing, and grinding (doping concentration: 15%Zn-8%Te, core–shell reaction time: 1 h:20 min:20 min, precursor ratio: 1:1:1). (<b>a</b>–<b>g</b>) SEM images of individual particles. (<b>h</b>–<b>l</b>) Ga, P, Zn, Te, and In element mapping of the particles shown in (<b>g</b>).</p>
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<p>XPS high-resolution spectra of the matrix elements in GaP:Zn/GaP/GaInP-GaP:Te/GaP (doping concentration: 15%Zn-8%Te, core–shell reaction time: 1 h:20 min:20 min, precursor ratio: 1:1:1) and pure GaP samples. (<b>a</b>) C 1s. (<b>b</b>) O 1s. (<b>c</b>) Ga 2p. (<b>d</b>) P 2p.</p>
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<p>XPS high-resolution spectra of the doped elements in GaP:Zn/GaP/GaInP-GaP:Te/GaP (doping concentration: 15%Zn-8%Te, core–shell reaction time: 1 h:20 min:20 min, precursor ratio: 1:1:1) and pure GaP samples. (<b>a</b>) Zn 2p. (<b>b</b>) Te 3d. (<b>c</b>) In 3d.</p>
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<p>Electroluminescence of GaP:Zn/GaP and GaP:Te/GaP samples. (<b>a</b>) Different doping concentrations of Zn and Te. (<b>b</b>) Electroluminescence intensity of 15%Zn-8%Te sample recorded at 600 nm with different voltage excitation. (<b>c</b>) Effect of grinding time on the electroluminescence performance of 15%Zn-8%Te samples.</p>
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<p>Electroluminescence of GaP:15%Zn/GaP/GaInP and GaP:8%Te/GaP samples with different core–shell precursor ratios and reaction times.</p>
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