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Search Results (917)

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36 pages, 2830 KiB  
Review
Real-Time, Continuous Monitoring of Tissue Chips as an Emerging Opportunity for Biosensing
by John S. Cognetti and Benjamin L. Miller
Sensors 2025, 25(16), 5153; https://doi.org/10.3390/s25165153 - 19 Aug 2025
Viewed by 370
Abstract
Tissue chips (TCs), otherwise known as organs-on-a-chip (OoC), organ chips (OCs), or microphysiological systems (MPS), are rapidly gaining prominence as an extension of or even replacement for traditional animal models of disease physiology. They also have recognized utility in the context of drug [...] Read more.
Tissue chips (TCs), otherwise known as organs-on-a-chip (OoC), organ chips (OCs), or microphysiological systems (MPS), are rapidly gaining prominence as an extension of or even replacement for traditional animal models of disease physiology. They also have recognized utility in the context of drug development: for example, data from TCs can now be submitted in place of some animal testing to the FDA. In principle, TCs are structured to allow measurement of any number of outputs that yield information about the tissue. However, to date, measurements made during experiments with TCs have been largely restricted to immunofluorescence microscopy and benchtop assays performed on media extracted from the cell culture within the device. With the development of biosensors that are sensitive and have an ever-shrinking footprint, on-board biosensing is now in the early stages of exploration. This review discusses the importance of tissue chips and the advances in sensing that will aid the complexity and utility of tissue chip research moving forward. We cover several sensing modalities, including electrical and optical sensing modes. Finally, challenges and opportunities for the future are discussed. Full article
(This article belongs to the Special Issue Feature Review Papers in Optical Sensors)
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14 pages, 784 KiB  
Article
Cytotoxicity of Bulk-Fill Composites on Stem Cells from Human Exfoliated Deciduous Teeth—An In Vitro Study
by Ralitsa Bogovska-Gigova, Nikolay Ishkitiev, Marina Miteva and Krasimir Hristov
Materials 2025, 18(16), 3863; https://doi.org/10.3390/ma18163863 - 18 Aug 2025
Viewed by 231
Abstract
Background: This study aimed to evaluate the cytotoxicity of bulk-fill composite materials compared to conventional compomers on stem cells from human exfoliated deciduous teeth. Methods: 90 standardized resin composite discs (4 mm thick, 4 mm diameter) were fabricated using a 3D-printed plate, comprising [...] Read more.
Background: This study aimed to evaluate the cytotoxicity of bulk-fill composite materials compared to conventional compomers on stem cells from human exfoliated deciduous teeth. Methods: 90 standardized resin composite discs (4 mm thick, 4 mm diameter) were fabricated using a 3D-printed plate, comprising four bulk-fill composites (SDR, Tetric EvoCeram Bulk-Fill, VisCalor Bulk, Cention-N) and one compomer (Dyract XP). Samples were polymerized per the manufacturer’s instructions and sterilized. Stem cells were isolated from the pulp of exfoliated primary teeth. Cells were cultured and exposed to extracts of the composite materials soaked in culture medium for 24 h. Cytotoxicity was assessed using the MTT colorimetric assay, measuring cell viability via mitochondrial activity, and the Annexin V assay, quantifying apoptosis and necrosis via flow cytometry. Statistical analysis was performed using ANOVA and Tukey post hoc tests. Results: All materials significantly reduced cell viability compared to the control (p < 0.05), with optical density values indicating high cytotoxicity. Tetric EvoCeram exhibited the lowest necrosis and apoptosis levels, while Dyract XP showed the highest necrosis. Statistical analysis revealed no significant cytotoxicity differences among most bulk-fill composites (p < 0.05). Conclusion: Bulk-fill composites and conventional compomer tested exhibit comparable and significant cytotoxic effects on stem cells from human exfoliated primary teeth pulp. While these materials offer clinical advantages in pediatric dentistry due to ease and speed of application, their use underscores the dilemma of balancing operative efficiency with biological safety, and their cytotoxic profiles should be taken into consideration prior to application. Full article
(This article belongs to the Special Issue Biomaterials for Restorative Dentistry)
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19 pages, 1567 KiB  
Article
Novel Carbazole–Thiazole Conjugates: Synthesis and Biophysical Characterization
by Beata Donarska, Klaudia Seklecka, Joanna Cytarska, Katarzyna Piechowska, Przemyslaw Ledwon, Sławomir Kula, Przemysław Krawczyk, Angelika Baranowska-Łączkowska and Krzysztof Z. Łączkowski
Int. J. Mol. Sci. 2025, 26(16), 7945; https://doi.org/10.3390/ijms26167945 - 18 Aug 2025
Viewed by 223
Abstract
This presented study depicts the synthesis of three novel carbazole–thiazole conjugates, thoroughly investigating their spectroscopic properties as well as evaluating their biological activity as tyrosinase inhibitors. Additionally, we investigated the possibility of using Concanavalin A (ConA) complexes with dyes from a theoretical point [...] Read more.
This presented study depicts the synthesis of three novel carbazole–thiazole conjugates, thoroughly investigating their spectroscopic properties as well as evaluating their biological activity as tyrosinase inhibitors. Additionally, we investigated the possibility of using Concanavalin A (ConA) complexes with dyes from a theoretical point of view, developing a promising protein-based strategy of delivery of dyes to the target cells. The tyrosinase inhibition assay showed that compounds K1 and K3 demonstrated higher activity than the kojic acid with IC50 values of 46 and 59 mM, respectively. Among the tested compounds, carbazole K3 exhibits the most pronounced nonlinear optical response due to its high electronic flexibility, strong solvatochromism, large excited-state dipole moments, and efficient intramolecular charge transfer. Additionally, all investigated carbazoles demonstrate high ability to form stable supramolecular complexes with ConA, which was confirmed using molecular docking studies. It was found experimentally and theoretically that the compound K3 has the best biophysical parameters, making it a promising candidate for potential diagnostic applications. Full article
(This article belongs to the Special Issue Spectroscopic Techniques in Molecular Sciences)
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18 pages, 3613 KiB  
Article
Early Biological Response to Poly(ε-caprolactone) PCL—Bioactive Glass Composites Obtained by 3D Printing as Bone Substitutes
by Alessandro Mosca Balma, Riccardo Pedraza, Ilaria Roato, Clarissa Orrico, Sara Meinardi, Stefano Bertinetti, Tullio Genova, Giovanna Gautier di Confiengo, Maria Giulia Faga, Donatella Duraccio, Giulio Malucelli, Marta Miola, Enrica Verné and Federico Mussano
Polymers 2025, 17(16), 2229; https://doi.org/10.3390/polym17162229 - 15 Aug 2025
Viewed by 480
Abstract
The increasing demand for smart bone substitutes has boosted the implementation of biomaterials possibly endowed with both pro-osteogenic and pro-angiogenic capabilities, among which bioactive glasses hold great potential. Hence, two Poly(ε-caprolactone) (PCL)-based composites were loaded at 10 wt.%, with either pristine (SBA3) or [...] Read more.
The increasing demand for smart bone substitutes has boosted the implementation of biomaterials possibly endowed with both pro-osteogenic and pro-angiogenic capabilities, among which bioactive glasses hold great potential. Hence, two Poly(ε-caprolactone) (PCL)-based composites were loaded at 10 wt.%, with either pristine (SBA3) or copper-doped (SBA3_Cu) silica-based bioactive glasses, through a solvent casting method with chloroform. Neat PCL was used as a control. Samples produced by 3D printing underwent SEM and EDX analyses, and the following were measured: tensile strength and hardness, surface roughness, ion release through ICP-OES, surface free energy, and optical contact angle. Adipose-derived mesenchymal stem cells (ASCs) and human microvascular endothelial cells (HMEC-1) were used to test the biocompatibility of the materials through cell adhesion, spreading, and viability assays. A significant improvement in tensile strength and hardness was observed especially with Cu-doped composites. Both SBA3 and SBA3_Cu added to the PCL favored the early adhesion and the proliferation of HMEC-1 after 3 and 7 days, while ASCs proliferated significantly the most on the SBA-containing composite, at all the time points. Cellular morphology analysis highlighted interesting adaptation patterns to the samples. Further biological characterizations are needed to understand thoroughly how specific bioactive glasses may interact with different cellular types. Full article
(This article belongs to the Section Polymer Composites and Nanocomposites)
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23 pages, 4468 KiB  
Article
Serrapeptase Eliminates Escherichia coli Biofilms by Targeting Curli Fibers, Lipopolysaccharides, and Phosphate Metabolism
by Georgios Katsipis, Michalis Aivaliotis and Anastasia A. Pantazaki
Microorganisms 2025, 13(8), 1875; https://doi.org/10.3390/microorganisms13081875 - 11 Aug 2025
Viewed by 548
Abstract
Escherichia coli biofilms are implicated in the development of persistent infections and increased antibiotic resistance, posing a significant challenge in clinical settings. These biofilms enhance bacterial survival by forming protective extracellular matrices, rendering conventional treatments less effective. Serrapeptase (SPT), a proteolytic enzyme, has [...] Read more.
Escherichia coli biofilms are implicated in the development of persistent infections and increased antibiotic resistance, posing a significant challenge in clinical settings. These biofilms enhance bacterial survival by forming protective extracellular matrices, rendering conventional treatments less effective. Serrapeptase (SPT), a proteolytic enzyme, has emerged as a potential anti-biofilm agent due to its ability to degrade biofilm components and disrupt bacterial adhesion. In this study, we report the inhibitory effect of SPT against E. coli biofilm and its effect on key virulence factors. In vitro assays, including crystal violet staining, optical and fluorescence microscopy, and viability measurements, revealed the dose-dependent inhibition of biofilm formation (IC50 = 14.2 ng/mL), reduced biofilm (−92%, 500 ng/mL) and planktonic viability (−45%, 500 ng/mL), and a marked loss of amyloid curli fibers. SPT treatment also lowered the levels of key virulence factors: cellular and secreted lipopolysaccharides (−76%, 8 ng/mL; −94%, 32 ng/mL), flagellin (−63%, 8 ng/mL), and peptidoglycan (−29%, 125 ng/mL). Mechanistically, SPT induced a phosphate-dysregulating response: secreted alkaline phosphatase activity rose (+70%, 125 ng/mL) while cellular DING/PstS proteins declined (−84%, 64 ng/mL), correlating strongly with biofilm inhibition. In silico docking further suggests direct interactions between SPT and the curli subunits CsgA and CsgB, potentially blocking fiber polymerization. Together, these findings position SPT as a powerful non-antibiotic biofilm disruptor against E. coli, offering a promising strategy to undermine bacterial persistence and resistance by targeting both structural matrix components and metabolic regulatory pathways. Full article
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14 pages, 5194 KiB  
Article
Drying-Induced Salt Deposition Patterns as a Tool for Label-Free Protein Quantification
by Arturo Patrone-Garcia, Miquel Avella-Oliver and Ángel Maquieira
Biosensors 2025, 15(8), 520; https://doi.org/10.3390/bios15080520 - 9 Aug 2025
Viewed by 316
Abstract
This work reports a label-free analytical strategy based on protein-induced modulation of salt crystallization patterns upon drying. This method relies on the consistent observation that protein-containing saline samples produce distinct salt deposition morphologies compared to protein-free controls. The work first demonstrates the concept [...] Read more.
This work reports a label-free analytical strategy based on protein-induced modulation of salt crystallization patterns upon drying. This method relies on the consistent observation that protein-containing saline samples produce distinct salt deposition morphologies compared to protein-free controls. The work first demonstrates the concept of this phenomenon and characterizes the structural features of the resulting salt patterns. Then, systematic experiments with different solution compositions, substrates, surface coatings, and protein types confirm the generality of this differential deposition behavior and its dependence on total protein concentration. Two complementary measurement approaches are evaluated: a custom laser-scattering setup for optical attenuation measurements and a digital image analysis method based on pixel intensity distributions. Both strategies enable quantitative protein detection in simple (casein) and complex (human serum) samples, offering good correlations between signal and concentration and detection limits in the range of 2–18 µg·mL−1 for digital image analysis and 162–205 µg·mL−1 for optical attenuation measurements. These findings introduce an appealing paradigm for protein quantification exploiting drying-mediated crystallization phenomena, with potential for simple and label-free bioanalytical assays. Full article
(This article belongs to the Special Issue Optical Sensors for Biological Detection)
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17 pages, 1848 KiB  
Article
Phenotypic Characterization of pilA, pilB, and pilD Mutants of Acinetobacter baumannii 5075: Impacts on Growth, Biofilm Formation, and Tazobactam Response
by Joel H. Salinas, Fatma Pinar Gordesli-Duatepe, Angelica Diaz-Sanchez and Nehal I. Abu-Lail
Antibiotics 2025, 14(8), 816; https://doi.org/10.3390/antibiotics14080816 - 9 Aug 2025
Viewed by 359
Abstract
Background/Objectives: The Type IV pilus assembly system in Acinetobacter baumannii is a major determinant of its pathogenicity, playing a role in surface-associated functions via the biogenesis of Type IV pili (T4P). Tazobactam (TAZ) is a well-characterized β-lactamase inhibitor, primarily used in combination with [...] Read more.
Background/Objectives: The Type IV pilus assembly system in Acinetobacter baumannii is a major determinant of its pathogenicity, playing a role in surface-associated functions via the biogenesis of Type IV pili (T4P). Tazobactam (TAZ) is a well-characterized β-lactamase inhibitor, primarily used in combination with β-lactam antibiotics such as piperacillin (PIP) to counteract bacterial resistance mechanisms. While A. baumannii resistance to β-lactam antibiotics has been well studied, the influence of T4P on its susceptibility to TAZ remains largely unexplored. For this reason, we investigated how multidrug-resistant A. baumannii 5075 (AB5075) responds to TAZ by assessing the roles of pilA, pilB, and pilD in bacterial growth and biofilm formation under direct TAZ exposure, with a focus on phenotypic characterization rather than molecular mechanisms. Methods: Bacterial growth kinetics were quantified by measuring the optical densities of cell suspensions and the colony forming units per volume (CFUs/mL) at different time intervals. Time-kill assays and microtiter dish biofilm formation assays were used to evaluate how effectively TAZ can inhibit growth and biofilm formation, respectively. Results: Time–kill assays confirmed that 32 µg/mL of TAZ inhibited growth in both wild-type (WT) and mutant strains, with the pilD mutant showing initial resistance before eventual inhibition. Biofilm assays showed that the pilA mutant had the highest biofilm formation at 8 h, surpassing the WT strain. A prolonged 32 µg/mL of TAZ exposure (24–36 h) significantly reduced biofilm production across all strains, with inhibition rates reaching 89% for the WT, 82% for the pilA mutant, 91% for the pilB mutant, and 86% for the pilD mutant. Conclusion: These findings deepen our understanding of the strain-specific roles of T4P components in growth and biofilm regulation in AB5075, and highlight the potential of TAZ as a therapeutic strategy against biofilm-associated infections. Full article
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19 pages, 3043 KiB  
Article
Enhancing Sensitivity of Commercial Gold Nanoparticle-Based Lateral Flow Assays: A Comparative Study of Colorimetric and Photothermal Approaches
by Jully Blackshare, Hansel A. Mina, Amanda J. Deering, Bartek Rajwa, J. Paul Robinson and Euiwon Bae
Sensors 2025, 25(16), 4904; https://doi.org/10.3390/s25164904 - 8 Aug 2025
Viewed by 348
Abstract
Lateral flow assays (LFAs) are extensively utilized in point-of-care diagnostics due to their affordability, simplicity, and rapid time-to-results. However, their low sensitivity remains a significant limitation, particularly for detecting foodborne pathogens at concentrations below regulatory thresholds. This study evaluated two distinct sensing modalities—photothermal [...] Read more.
Lateral flow assays (LFAs) are extensively utilized in point-of-care diagnostics due to their affordability, simplicity, and rapid time-to-results. However, their low sensitivity remains a significant limitation, particularly for detecting foodborne pathogens at concentrations below regulatory thresholds. This study evaluated two distinct sensing modalities—photothermal speckle imaging and colorimetric line intensity analysis—for their potential to enhance the sensitivity of commercially available LFAs. Photothermal imaging quantified refractive index shifts induced by plasmonic heating of gold nanoparticles, while colorimetric analysis used smartphone-acquired images processed with machine learning. The photothermal method achieved a limit of detection (LOD) of 2.13 × 105 CFU/mL, while the colorimetric approach, using a logistic regression model with LASSO regularization, achieved an LOD of 105 CFU/mL. While both approaches demonstrated detection thresholds comparable to traditional visual interpretation, the colorimetric method provided an added advantage by enabling quantitative prediction of bacterial concentration through regression modeling. With further optimization of each sensing method, these findings demonstrate the feasibility of improving unmodified commercial LFAs through optical and computational enhancements, offering a promising pathway toward the development of portable biosensing systems for real-time food safety monitoring. Full article
(This article belongs to the Special Issue Feature Papers in Optical Sensors 2025)
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21 pages, 6387 KiB  
Article
Carbon Dot-Enhanced Doxorubicin Liposomes: A Dual-Functional Nanoplatform for Cancer Therapy
by Corina-Lenuta Logigan, Cristian Peptu, Corneliu S. Stan, Gabriel Luta, Crina Elena Tiron, Mariana Pinteala, Aleksander Foryś, Bogdan Simionescu, Constanta Ibanescu, Adrian Tiron and Catalina A. Peptu
Int. J. Mol. Sci. 2025, 26(15), 7535; https://doi.org/10.3390/ijms26157535 - 4 Aug 2025
Viewed by 464
Abstract
Liposomes (LPs) represent one of the most effective nanoscale platforms for drug delivery in cancer therapy due to their favorable pharmacokinetic and various body tissue compatibility profiles. Building on recent findings showing that carbon dots derived from N-hydroxyphthalimide (CDs-NHF) possess intrinsic antitumor activity, [...] Read more.
Liposomes (LPs) represent one of the most effective nanoscale platforms for drug delivery in cancer therapy due to their favorable pharmacokinetic and various body tissue compatibility profiles. Building on recent findings showing that carbon dots derived from N-hydroxyphthalimide (CDs-NHF) possess intrinsic antitumor activity, herein, we investigate the possibility of preparing complex nano-platforms composed of LPs encapsulating CDs-NHF and/or doxorubicin (DOX) for breast and lung cancer. Various LP formulations were prepared and characterized using Cryo-TEM and Cryo-SEM for morphological analysis, while zeta potential and fluorescence assessments confirmed their stability and optical properties. Cellular effects were evaluated through immunofluorescence microscopy and proliferation assays. LPs-CDs-NHF significantly reduced cancer cell viability at lower concentrations compared to free CDs-NHF, and this effect was further amplified when combined with doxorubicin. Mechanistically, the liposomal formulations downregulated key signaling molecules including pAKT, pmTOR, and pERK, indicating the disruption of cancer-related pathways. These findings suggest that LPs containing CDs-NHF, either alone or in combination with DOX, exhibit synergistic antitumor activity and hold strong promise as multifunctional nanocarriers for future oncological applications. Full article
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21 pages, 2189 KiB  
Article
Surface Modification, Characterization, and Cytotoxicity of Ti-6Al-4V Alloy Enriched by EDM Process
by Bárbara A. B. dos Santos, Elaine C. S. Corrêa, Wellington Lopes, Liszt Y. C. Madruga, Ketul C. Popat, Roberta M. Sabino and Hermes de Souza Costa
Appl. Sci. 2025, 15(15), 8443; https://doi.org/10.3390/app15158443 - 30 Jul 2025
Viewed by 497
Abstract
This study investigates the surface modification of Ti-6Al-4V alloy through the electrical discharge machining (EDM) process to improve its suitability for orthopedic and dental implant applications. The analysis focused on evaluating the morphological, wettability, roughness, hardness, and biocompatibility properties of the modified surfaces. [...] Read more.
This study investigates the surface modification of Ti-6Al-4V alloy through the electrical discharge machining (EDM) process to improve its suitability for orthopedic and dental implant applications. The analysis focused on evaluating the morphological, wettability, roughness, hardness, and biocompatibility properties of the modified surfaces. Samples were subjected to different dielectric fluids and polarities during EDM. Subsequently, optical microscopy, roughness measurements, Vickers microhardness, contact angle tests, and in vitro cytotoxicity assays were performed. The results demonstrated that EDM processing led to the formation of distinct layers on the sample surfaces, with surface roughness increasing under negative polarity by up to ~304% in Ra and 305% in Rz. Additionally, wettability measurements indicated that the modified surfaces presented a lower water contact angle, which suggests enhanced hydrophilicity. Moreover, the modified samples showed a significant increase in Vickers microhardness, with the highest value reaching 1520 HV in the recast layer, indicating improvements in the mechanical properties. According to ISO 10993-5, all treated samples were classified as non-cytotoxic, presenting RGR values above 75%, similar to the untreated Ti-6Al-4V alloy. Therefore, it is concluded that surface modification through the EDM process has the potential to enhance the properties and safety of biomedical implants made with this alloy. Full article
(This article belongs to the Special Issue Titanium and Its Compounds: Properties and Innovative Applications)
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41 pages, 11320 KiB  
Review
Electrochemical Biosensors Driving Model Transformation for Food Testing
by Xinxin Wu, Zhecong Yuan, Shujie Gao, Xinai Zhang, Hany S. El-Mesery, Wenjie Lu, Xiaoli Dai and Rongjin Xu
Foods 2025, 14(15), 2669; https://doi.org/10.3390/foods14152669 - 29 Jul 2025
Viewed by 707
Abstract
Electrochemical biosensors are revolutionizing food testing by addressing critical limitations of conventional strategies that suffer from cost, complexity, and field-deployment challenges. Emerging fluorescence and Raman techniques, while promising, face intrinsic drawbacks like photobleaching and matrix interference in opaque or heterogeneous samples. In contrast, [...] Read more.
Electrochemical biosensors are revolutionizing food testing by addressing critical limitations of conventional strategies that suffer from cost, complexity, and field-deployment challenges. Emerging fluorescence and Raman techniques, while promising, face intrinsic drawbacks like photobleaching and matrix interference in opaque or heterogeneous samples. In contrast, electrochemical biosensors leverage electrical signals to bypass optical constraints, enabling rapid, cost-effective, and pretreatment-free analysis of turbid food matrices. This review highlights their operational mechanisms, emphasizing nano-enhanced signal amplification (e.g., Au nanoparticles and graphene) and biorecognition elements (antibodies, aptamers, and molecularly imprinted polymers) for ultrasensitive assay of contaminants, additives, and adulterants. By integrating portability, scalability, and real-time capabilities, electrochemical biosensors align with global food safety regulations and sustainability goals. Challenges in standardization, multiplexed analysis, and long-term stability are discussed, alongside future directions toward AI-driven analytics, biodegradable sensors, and blockchain-enabled traceability, ultimately fostering precision-driven, next-generation food safety and quality testing. Full article
(This article belongs to the Section Food Analytical Methods)
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13 pages, 2153 KiB  
Article
Interaction of MG63 Human Osteosarcoma-Derived Cells on S53P4 Bioactive Glass: An In Vitro Study
by Valentin Schmidt, Beáta Polgár, Vanda Ágnes Nemes, Tímea Dergez, László Janovák, Péter Maróti, Szilárd Rendeki, Kinga Turzó and Balázs Patczai
J. Funct. Biomater. 2025, 16(8), 275; https://doi.org/10.3390/jfb16080275 - 29 Jul 2025
Viewed by 664
Abstract
Bioactive glass materials have been used for decades in orthopedic surgery, traumatology, and oral and maxillofacial surgery to repair bone defects. This study aimed to evaluate in vitro the survival and proliferation of MG63 human osteosarcoma-derived cells on S53P4 bioactive glass (BonAlive® [...] Read more.
Bioactive glass materials have been used for decades in orthopedic surgery, traumatology, and oral and maxillofacial surgery to repair bone defects. This study aimed to evaluate in vitro the survival and proliferation of MG63 human osteosarcoma-derived cells on S53P4 bioactive glass (BonAlive® granules). Microscopic visualization was performed to directly observe the interactions between the cells and the material. Osteoblast-like cells were examined on non-adherent test plates, on tissue culture (TC)-treated plates and on the surface of the bioglass to assess the differences. Cell survival and proliferation were monitored using a CCK-8 optical density assay. Comparing the mean OD of MG63 cells in MEM on TC-treated plates with cells on BG, we detected a significant difference (p < 0.05), over each time of observation. The sustained cell proliferation confirmed the non-cytotoxic property of the bioglass, as the cell number increased continuously at 48, 72, 96, and 168 h and even did not plateau after 168 h. Since the properties of bioglasses can vary significantly depending on their composition and environment, a thorough characterization of their biocompatibility is crucial to ensure their effective and appropriate application—for example, during hip and knee prosthesis insertion. Full article
(This article belongs to the Section Bone Biomaterials)
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18 pages, 6694 KiB  
Article
Effects of a ROCK Inhibitor on Retinal Ganglion Cells In Vivo and In Vitro
by Wanjing Chen, Yoko Iizuka, Fumihiko Mabuchi and Kenji Kashiwagi
J. Clin. Med. 2025, 14(15), 5344; https://doi.org/10.3390/jcm14155344 - 29 Jul 2025
Viewed by 383
Abstract
Objective: To investigate the neuroprotective effects of a Rho-associated kinase (ROCK) inhibitor on retinal ganglion cells (RGCs) in vitro and in vivo. Methods: For in vivo studies, a unilateral optic nerve crush mouse model was established. Then, 100 mM Y-27632 (a [...] Read more.
Objective: To investigate the neuroprotective effects of a Rho-associated kinase (ROCK) inhibitor on retinal ganglion cells (RGCs) in vitro and in vivo. Methods: For in vivo studies, a unilateral optic nerve crush mouse model was established. Then, 100 mM Y-27632 (a ROCK inhibitor) or saline was applied to the experimental eyes once a day for 14 days. The effects of the ROCK inhibitor were evaluated by counting the surviving RGCs in the enucleated flat retina tissues and measuring the inner retinal thickness using optical coherence tomography (OCT), the amplitude of the electroretinogram (ERG), and the change in intraocular pressure (IOP). For the in vitro study, RGCs were isolated from five-day-old mice using a modified immunopanning method with magnetic beads. The isolated RGCs were incubated for 72 h with various concentrations of Y-27632, after which TUNEL assays were performed to determine the number of surviving RGCs. Results: Y-27632 has neuroprotective effects, as it significantly increased the number of surviving RGCs by approximately 6.3%. OCT and ERG data also revealed that Y-27632 induced neuroprotective effects in vivo; furthermore, Y-27632 reduced IOP by approximately 18.3%. The in vitro study revealed the dose-dependent neuroprotective effects of Y-27632, with the highest dose of Y-27632 (1000 nM) increasing the RGC survival rate after 72 h of incubation compared with that of the control. Conclusions: The ROCK inhibitor Y-27632 may exert some neuroprotective effects on RGCs when it is used as an eye drop through an IOP-independent mechanism. Full article
(This article belongs to the Section Ophthalmology)
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24 pages, 1138 KiB  
Review
Eyes Wide Open: Assessing Early Visual Behavior in Zebrafish Larvae
by Michela Giacich, Maria Marchese, Devid Damiani, Filippo Maria Santorelli and Valentina Naef
Biology 2025, 14(8), 934; https://doi.org/10.3390/biology14080934 - 24 Jul 2025
Viewed by 467
Abstract
Early diagnosis is critical for the effective management of neurodegenerative disorders, and retinal alterations have emerged as promising early biomarkers due to the retina’s close developmental and functional link to the brain. The zebrafish (Danio rerio), with its rapid development, transparent embryos, and [...] Read more.
Early diagnosis is critical for the effective management of neurodegenerative disorders, and retinal alterations have emerged as promising early biomarkers due to the retina’s close developmental and functional link to the brain. The zebrafish (Danio rerio), with its rapid development, transparent embryos, and evolutionarily conserved visual system, represents a powerful and versatile model for studying retinal degeneration. This review discusses a range of behavioral assays—including visual adaptation, motion detection, and color discrimination—that are employed to evaluate retinal function in zebrafish. These methods enable the detection of subtle visual deficits that may precede overt anatomical damage, providing a non-invasive, efficient strategy for early diagnosis and high-throughput drug screening. Importantly, these behavioral tests also serve as sensitive functional readouts to evaluate the efficacy of pharmacological treatments over time. Compared to traditional murine models, zebrafish offer advantages such as lower maintenance costs, faster development, optical transparency for live imaging, and ethical benefits due to reduced use of higher vertebrates. However, variability in experimental protocols highlights the need for standardization to ensure reliability and reproducibility. Full article
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18 pages, 2560 KiB  
Article
In Vitro Insights into the Anti-Biofilm Potential of Salmonella Infantis Phages
by Jan Torres-Boncompte, María Sanz-Zapata, Josep Garcia-Llorens, José M. Soriano, Pablo Catalá-Gregori and Sandra Sevilla-Navarro
Antibiotics 2025, 14(8), 744; https://doi.org/10.3390/antibiotics14080744 - 24 Jul 2025
Viewed by 521
Abstract
Background/Objectives: As bacteriophage-based strategies to control bacterial pathogens continue to gain momentum, phage therapy is increasingly being explored across various fields. In the poultry industry, efforts to minimize the public health impact of Salmonella have spurred growing interest in phage applications, particularly [...] Read more.
Background/Objectives: As bacteriophage-based strategies to control bacterial pathogens continue to gain momentum, phage therapy is increasingly being explored across various fields. In the poultry industry, efforts to minimize the public health impact of Salmonella have spurred growing interest in phage applications, particularly as prophylactic and disinfecting agents. Although the disinfecting potential of bacteriophages has been recognized, in-depth studies examining their efficacy under varying environmental conditions remain limited. This study focused on evaluating the effectiveness of bacteriophages as disinfecting agents against biofilm-forming Salmonella Infantis under different environments. Methods: A comprehensive screening of biofilm-producing strains was conducted using Congo Red Agar and 96-well plate assays. Two strains with distinct biofilm-forming capacities were selected for further analysis under different environmental conditions: aerobic and microaerobic atmospheres at both 25 °C and 37 °C. The resulting biofilms were then treated with four phage preparations: three individual phages and one phage cocktail. Biofilm reduction was assessed by measuring optical density and CFU/well. Additionally, scanning electron microscopy was used to visualize both untreated and phage-treated biofilms. Results: The results demonstrated that all S. Infantis strains were capable of forming biofilms (21/21). All three phage candidates exhibited biofilm-disrupting activity and were able to lyse biofilm-embedded Salmonella cells. Notably, the lytic efficacy of the phages varied depending on environmental conditions, highlighting the importance of thorough phage characterization prior to application. Conclusions: These findings underscore that the effectiveness of bacteriophages as surface disinfectants can be significantly compromised if inappropriate phages are used, especially in the presence of biofilms. Full article
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