Search Results (2,722)

The physiological relevance of in vitro models is limited because conventional two-dimensional cell culture systems are unable to replicate the structural and functional complexity of native tissues. Extracellular matrix (ECM)-mimetic hydrogels have become important platforms for tissue engineering applications. This work developed hybrid hydrogels that mimic important biochemical and mechanical characteristics of cardiac tissue by combining decellularized bovine pericardium-derived (dBP) ECM, gellan gum (GG), and spermine (SPM). Although dBP offers tissue-specific biological cues, processing compromises its mechanical integrity. This limitation was overcome by adding GG, whose ionic gelation properties were optimized using DMEM and SPM. The hydrogels’ mechanical, biological, physicochemical, and structural characteristics were all evaluated. Under physiologically simulated conditions, the formulations showed quick gelation and long-term stability; scanning electron microscopy revealed an interconnected, ECM-like porous microarchitecture. While uniaxial compression testing provided Young’s modulus values comparable to native myocardium, rheological analysis revealed a concentration-dependent increase in storage modulus with increasing SPM content. H9C2 cardiomyoblasts were used in cytocompatibility studies to confirm that cell viability, morphology, and cytoskeletal organization were all preserved. All of these findings support the potential application of dBP−GG−SPM hydrogels in advanced in vitro cardiac models by showing that they successfully replicate important characteristics of cardiac ECM. Full article

There is a growing pressure for further breast cancer treatment development to improve patient quality of life and clinical effectiveness while reducing drug resistance and side effects. Clinical research, however, remains both expensive and time-consuming, highlighting the importance of advancing preclinical studies. The ability of a model to accurately replicate the biological environment is essential for generating reliable data. Although research on two-dimensional in vitro cell culture systems has been conducted for over a century, their limited complexity hinders further development of cancer treatments. Consequently, the response to tested medications is artificial, since some drugs appear to be effective only in the two-dimensional model and not in clinical settings. A multicellular spheroid (MCS) is a three-dimensional assembly of cancer cells, used in oncology to study cancer biology and drug sensitivity. MCSs serve as valuable in vitro models in preclinical drug development. They can also be used in a clinical setting as a personalized tool for drug susceptibility screening. The objective of this review is to highlight new possibilities for improved anticancer drug screening and the development of personalized treatment regimens with multicellular spheroids. Full article

Background: gemcitabine is a cytidine analog and major anticancer drug functioning as an antimetabolite. However, its administration by systemic route is accompanied by “burst” and side effects. To limit this, drugs are encapsulated in matrices; metal–organic frameworks (MOFs) are coordination polymers with strong potential for drug encapsulation and delayed release. Methods: mechano-chemical synthesis of solid-state binary complex lag(CYCU-3)(Gem) is described from aluminum MOF (Al-MOF) CYCU-3 and gemcitabine free base (Gem). Synthesis is conducted by liquid-assisted grinding (LAG) with dimethyl sulfoxide (DMSO) followed by its outgassing. The alternative “dry” synthesis results in dry(CYCU-3)(Gem). Materials were characterized by FTIR spectroscopy and XRD, and delayed Gem release was tested to phosphate buffered saline (PBS) at 37 °C. The in vitro toxicity to pancreatic cancer PANC−1 and healthy cells hTERT−HPNE E6/E7/K−RasG12D was assessed by fluorometric assay. Results: in lag(CYCU-3)(Gem) interactions MOF-drug are via non-covalent bonds at O-H and COO⁻ groups of CYCU-3 as found by FTIR marker peak shifts and crystal structure is retained, while dry(CYCU-3)(Gem) shows significant amorphization and loss of functional groups. The lag(CYCU-3)(Gem) but not dry(CYCU-3)(Gem) shows delayed Gem release for 6000 min. The suppression of PANC−1 cells by lag(CYCU-3)(Gem) is time-dependent and it correlates with delayed Gem release. For the first time, a concept of ternary stoichiometric complex lag(CYCU-3)₁(Gem)₁(CIT)₂ is tested that also contains natural organic compound citronellol (CIT), and its structure, bonding and release of Gem are compared to those of binary complex. Bonding is at the O-H groups of CYCU-3 and this complex shows delayed Gem release. Conclusions: binary and ternary complexes of Gem with CYCU-3 yield delayed release and cytotoxicity. LAG is promising for synthesis of solid-state complexes of gemcitabine for delayed release and time-dependent suppression of cancer cells. Full article

Clinoptilolite is a zeolite with a microporous structure that enables ion exchange, molecular sieving, and adsorption, conferring detoxifying, antioxidant, and anti-inflammatory properties. These properties have applications in food, medicine, catalysis, and environmental remediation. This study evaluated the safety of the lab-made Clinoptilolite as a potential food ingredient through a 90-day repeated-dose toxicity study in male and female Sprague Dawley rats. The test substance was administered via oral gavage at doses of 0, 5, 10, and 15 mg/kg bw/day, followed by a 28-day recovery period. In addition, genotoxicity was assessed using the Ames test, in vitro chromosomal aberration assay, and an in vivo micronucleus test. All studies were conducted in accordance with OECD and FDA guidelines. Results showed no adverse systemic, genotoxic, or irreversible effects at any dose, with minor clinical variations being incidental and reversible. Genotoxicity tests confirmed no mutagenic or clastogenic potential. Overall, the lab-made Clinoptilolite evaluated in this investigation was well tolerated, non-toxic, and showed no evidence of treatment-related toxicity at the doses tested. These findings provide supportive evidence for its consideration toward a Generally Recognized as Safe (GRAS) determination. Full article

Amyotrophic lateral sclerosis (ALS) is the most common motor neuron disease, marked by progressive degeneration of upper and lower motor neurons. Clinically, genetically, and pathologically heterogeneous, ALS poses a major challenge for disease modeling and therapeutic translation. Over the past two decades, induced pluripotent stem cells (iPSCs) have reshaped our understanding of ALS pathogenesis and emerged as a promising translational platform for therapy development. ALS modeling has further expanded with the advent of three-dimensional systems, including ALS-on-chip platforms and organoid models, which better capture cell–cell interactions and tissue-level phenotypes. Despite these advances, effective disease-modifying therapies remain elusive. Recent clinical trial setbacks highlight the need for improved trial design alongside robust, translational iPSC models that can better predict therapeutic response. Nonetheless, the outlook is promising as large iPSC patient cohorts, quantitative phenotyping combined with genetically informed patient stratification, and reverse translational research are beginning to close the gap between in vitro discovery and clinical testing. In this review, we summarize the major advances in iPSC technology and highlight key iPSC-based studies of sporadic ALS. We further discuss emerging examples of iPSC-informed therapeutic strategies and outline the challenges associated with translating iPSC-derived mechanistic insights and pharmacological findings into successful clinical therapies. Full article

The interpretation and diagnosis of central sleep apnea in pediatrics by nocturnal polysomnography is challenging due to its technical complexity, which involves the simultaneous recording of multiple physiological parameters related to sleep and wakefulness. Furthermore, the unfamiliar environment of a sleep laboratory can hinder sleep evaluation, and diagnostic backlogs are common due to restricted capacity at specialist tertiary centers. The ability to undertake home sleep studies in a familiar environment using simple, robust, and low-cost technology is attractive. The potential to repurpose the PneumoWave biosensor, a UKCA Class 1 device, registered as an accelerometer-based monitoring device that is intended to capture and store chest motion data continuously over a period of time for retrospective analysis, was explored in an in vitro model of central sleep apnea. The PneumoWave system contains a biosensor (PW010), which was able to record simulated apnea episodes of 5 to 20 s across physiologically relevant pediatric breathing rates using an in vitro manikin model and manual annotation. The findings confirm that the PneumoWave biosensor could be a useful technology to support home sleep apnea testing and warrant further exploration. Full article

Coxsackieviruses B are single-stranded RNA viruses belonging to the Enterovirus genus and are associated with various clinical outcomes, ranging from acute infections to chronic diseases, such as type 1 diabetes (T1D). It was previously shown that inoculation of Swiss albino mice with CVB4 by the intraperitoneal route induced both anti-CVB4 neutralizing and enhancing activities of serum. This study aimed to investigate the humoral immune response of mice inoculated with CVB4 by the mucosal route. Mice were inoculated orally or intranasally with CVB4, and the anti-CVB4 neutralizing activity of serum and saliva was assessed by a cell culture neutralization assay. Anti-enterovirus (EV) IgG and IgA antibodies were detected in serum and saliva, respectively, by ELISA. The serum-dependent enhancement of CVB4 infection in cultures of murine splenocytes was evaluated by detecting intracellular viral RNA using RT-qPCR. At day 45 post-inoculation, an anti-CVB4 neutralizing activity, the extent of which depends on the amount of inoculated infectious particles, was detected in the serum of mice exposed orally or intranasally. An increase in anti-CVB4 neutralizing activity was observed in the saliva of mice inoculated orally or intranasally during the follow-up. Oral or intranasal inoculation of CVB4 induced a systemic IgG and mucosal IgA response. In addition, serum from these mice harbored an anti-CVB4 enhancing activity in vitro. These data indicate that Swiss albino mice exposed to CVB4 via the mucosal route constitute a potentially useful model for testing strategies to promote the production of protective mucosal and systemic anti-CVB4 antibodies and for verifying whether or not enhanced antibodies are produced. Full article

The purpose of this study was to characterise the effect of cranberry (Vaccinium macrocarpon) juice on unicellular and multicellular systems, specifically food spoilage yeasts (Wickerhamomyces anomalus, Dekkera bruxellensis and Rhodotorula mucilaginosa) and intestinal cells (IEC-6 and Caco-2 cells). The effects of both raw cranberry juice and juice digested in vitro were investigated. The juices were analysed for polyphenol content using high-performance liquid chromatography coupled with mass spectrometry. The cranberry juice was evaluated for its impact on yeast surface hydrophobicity and anti-adhesive action using the MATH test and luminometry/microscopy, respectively. We also assessed the effects of raw and digested cranberry juices on IEC-6 and Caco-2 cells by measuring cell viability, metabolic modulation, genotoxicity, and antioxidant activity. Chromatographic analysis of the raw cranberry juice revealed the presence of diverse bioactive compounds, identified as hydroxybenzoic and hydroxycinnamic acids, flavonols, and anthocyanins. After digestion, the cranberry juice remained a rich source of phenolic acids. The yeast strain R. mucilaginosa was characterised by the highest hydrophobicity and adhesive abilities, but cell adhesion in the presence of raw cranberry juice was several times lower for all the tested strains. Both tested cranberry juices reduced ROS levels and were well tolerated by intestinal epithelial cells, without significant cytotoxic or genotoxic effects. Our findings provide new insights into the safety of using cranberry juice across unicellular and multicellular systems. However, further validation in real-world settings is necessary before practical applications. Full article

Strategies to suppress methanogenesis must preserve the functional integrity of the rumen microbial ecosystem. Essential oils (EOs) have emerged as promising modulators of rumen microbial function, though their responses vary widely with chemical structure and inclusion level. This study evaluated the efficacy of selected EOs using detailed in vitro fermentation assays. Nine EOs—cinnamon, lavender, garlic (GAR), lemongrass (LEG), peppermint (PPM), eucalyptus, coriander, oregano, and ginger (GIN)—were evaluated for their effects on rumen fermentation and methane (CH₄) production using a 24 h in vitro batch culture system. Eight EOs were tested at two doses (Low and High) specific to each EO, while GIN was evaluated at a single dose. All treatments were incubated in a rumen fluid–buffer mix (1:1 for fermentation parameters and 1:4 for gas and CH₄ measurements) with a 55:45 forage-to-concentrate substrate (pH 6.9). Overall treatment effects were significant for all measured fermentation parameters (p < 0.01). Most treatments reduced total gas production, CH₄ emissions, and CH₄/total gas ratios compared with the control (p < 0.05), although several responses were dose-dependent or directly divergent. Essential oils showed clear, composition-dependent responses: non-terpenoid EOs produced the strongest but also the most variable antimethanogenic effects, with GAR, particularly at the lower dose, consistently achieving the greatest CH₄ inhibition while maintaining a favorable fermentation pattern. Conversely, terpenoid-based EOs induced moderate, dose-responsive CH₄ reductions with minimal effects on overall fermentation. At the higher dose, PPM suppressed CH₄ without altering major volatile fatty acid (VFA) patterns aside from increases in valerate and branched-chain VFA, whereas LEG reduced CH₄ only when accompanied by marked fermentation depression. Monensin validated its role as an effective positive control. Overall, GAR, characterized by sulfur-based bioactives, emerged as the most effective candidate for CH₄ mitigation under the tested in vitro conditions, highlighting the importance of chemical composition and inclusion level in determining efficacy and reinforcing the need for in vivo validation. Full article

Hyaluronic acid (HA) is a natural biopolymer widely used in wound dressings for its supportive role in the healing process. In this study, we investigated the occurrence of microorganisms in chronic wounds and evaluated the antimicrobial activity of newly synthesized HA–Starch-based materials enriched with acidifying agents. Microbial isolates obtained from chronic wounds were tested for susceptibility using the agar diffusion method. The prepared materials exhibited significant antimicrobial activity against both reference strains and multidrug-resistant clinical isolates. Further characterization by scanning electron microscopy and elemental analysis confirmed uniform microfiber morphology and the expected elemental composition of the fibers. Cytotoxicity assessments performed using the xCELLigence system demonstrated the potential safety of developed materials. Overall, the results indicate that HA–Starch-based materials containing acidifying compounds exhibit strong in vitro antimicrobial activity against chronic-wound isolates, supporting their potential for further evaluation in wound care applications. Full article

The rapid evolution of coronaviruses (CoVs) requires researchers to develop specific yet broad-spectrum detection methods to monitor their constant genomic changes. The goal of the present study was to establish a current pan-coronavirus RT-PCR system capable of detecting a wide variety of CoVs and useful for the investigation of virus diversity and host spectrum. For optimization, one-step and two-step nested RT-PCRs with three RT enzymes were examined, amplifying a ~600 bp long product of the RNA-dependent RNA polymerase. As templates, the in vitro transcribed RNA of ten pathogenic CoVs (SARS-CoV, SARS-CoV-2, NL-63, OC43, feline CoV, porcine epidemic diarrhea virus or PEDV, transmissible gastroenteritis virus or TGEV, canine CoV, bat CoV, and infectious bronchitis virus) were applied instead of the often-used DNA standards. A limit of detection of 5–50 copies/reaction was achieved with a random hexamer-primed two-step RT-PCR and a touchdown cycling profile, representing a lower detection limit and higher specificity compared to previously published primer sets. Swine origin pooled samples (n = 121), collected from apparently healthy herds in Hungary, were tested with the novel RT-PCR system. Sequences of porcine respiratory CoV/TGEV and porcine hemagglutinating encephalomyelitis virus were identified in 24 oral fluid and nasal swab pools, demonstrating the circulation of these viruses in this country, as well as the suitability of the new PCR for their detection. The results highlighted the importance of adequate RT enzyme selection and the use of RNase inhibitors in sample preparation and conservation. Full article

Soybean root rot caused by Rhizoctonia solani is a yield-limiting disease in Northeast China, particularly under continuous monoculture and cool climatic conditions. Despite its agronomic impact, the epidemiology and fungicide resistance profile of the pathogen remain inadequately characterized. In this study, a comprehensive survey conducted in Heilongjiang Province yielded 990 pathogenic isolates belonging to 11 fungal species. Among them, 55 strains were identified as R. solani based on combined morphological and molecular analyses. These isolates induced typical symptoms of root and stem browning with constriction. Pathogenicity tests on 30 R. solani isolates indicated that 83.3% were highly pathogenic. The pathogen exhibited a distinct geographic distribution, with the highest percentage of pathogen isolation recorded in Jiamusi (26.6%), which accounted for 61.8% of all R. solani isolates. In vitro fungicide sensitivity assays demonstrated that fludioxonil and prochloraz were highly effective (EC₅₀ < 0.0050 µg·mL⁻¹), whereas resistance was observed to tebuconazole, difenoconazole, pyraclostrobin, and carbendazim. Pot experiments confirmed that fludioxonil seed treatment (15 g a.i./100 kg seeds) provided superior control efficacy (63.07%) compared to prochloraz (46.85%). These findings establish R. solani as a dominant causal agent of soybean root rot in the region and support the prioritized use of fludioxonil for sustainable disease management. By elucidating the pathogenicity, distribution, and resistance patterns of R. solani, this study provides critical insights for controlling soybean root rot in cold-climate production systems and facilitates the development of targeted management strategies. Full article

Background: Solid microneedles (MNs) are effective transdermal delivery devices but are commonly fabricated from metallic or non-biodegradable materials, raising concerns related to sustainability, waste management, and processing constraints. This study aimed to evaluate the suitability of the biodegradable biopolyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) (PHBHVHHx) as a structuring material for solvent-free fabrication of solid MN arrays and to assess their mechanical performance, insertion capability, and drug delivery potential. Methods: PHBHVHHx MN arrays were fabricated by solvent-free micromolding at 200 °C. The resulting MNs were morphologically characterized by scanning electron microscopy. Mechanical properties were assessed by axial compression testing, and insertion performance was evaluated using a multilayer Parafilm skin simulant model. Diclofenac sodium was used as a model drug and applied via surface coating using a FucoPol-based formulation. In vitro drug release was assessed in phosphate-buffered saline under sink conditions and quantified by UV–Vis spectroscopy. Results: PHBHVHHx MN arrays consisted of sharp, well-defined conical needles (681 ± 45 µm length; 330 µm base diameter) with micro-textured surfaces. The MNs withstood compressive forces up to 0.25 ± 0.03 N/needle and achieved insertion depths of approximately 396 µm in the Parafilm model. Drug-coated MNs retained adequate mechanical integrity and exhibited a rapid release profile, with approximately 73% of diclofenac sodium released within 10 min. Conclusions: The results demonstrate that PHBHVHHx is a suitable biodegradable thermoplastic for the fabrication of solid MN arrays via a solvent-free process. PHBHVHHx MNs combine adequate mechanical performance, reliable insertion capability, and compatibility with coated drug delivery, supporting their potential as sustainable alternatives to conventional solid MN systems. Full article

Background: Combating antimicrobial resistance and developing dressings that match all aspects of wound healing will always be challenging. Methods: In this study, hydrogel membranes composed of sodium alginate (SA), polyvinyl alcohol (PVA), and Pluronic-f-127 (F-127) loaded with colistin (C) were formulated. The formulations were divided into two groups: group 1 (SA-PVA-C) and group 2 (SA-PVA-F127-C). Results: The membranes were characterized using multiple techniques, which confirmed component compatibility, physical cross-linking, an amorphous structure, and suitable surface morphology with acceptable porosity. Mechanical testing showed that both groups were suitable for wound-dressing applications. Differences in drug release across media (water, normal saline, and phosphate) were non-significant (p value > 0.05). Drug-loaded membranes (n = 3) from both groups showed antibacterial activity against multidrug-resistant Gram-negative Pseudomonas aeruginosa (ZOI = 20.33 ± 2.51 mm, 21.66 ± 2.08 mm). Conclusions: Overall, the developed hydrogel membranes (both group 1 and group 2) demonstrated promising in vitro potential as colistin delivery systems for wound infection management. Full article

Background/Objectives: Assessing the bioavailability of nutrients and bioactive compounds in vitro commonly relies on coupling standardized gastrointestinal digestion models with intestinal epithelial cell systems. However, digests produced using static digestion protocols such as INFOGEST often impair epithelial barrier integrity, limiting their direct application to intestinal models and reducing reproducibility across studies. Methods: This work systematically compared five commonly used digest conditioning strategies, including acidification, centrifugation, rapid freezing, and ultrafiltration using 10 kDa and 3 kDa molecular weight cut-off membranes, to identify the approach that best preserves intestinal epithelial viability and barrier function while enabling exposure at physiologically relevant concentrations. INFOGEST digests of yogurt were initially evaluated, followed by validation using biscuit and canned mackerel digests. Cell viability and monolayer integrity were assessed in differentiated Caco-2 cells using MTT assay and transepithelial electrical resistance (TEER) measurements. Results: Among the tested approaches, ultrafiltration using 3 kDa membranes consistently preserved epithelial viability and barrier integrity at a 1:10 dilution across all food matrices, whereas other conditioning methods failed to maintain TEER despite acceptable cell viability. At lower dilutions, food-dependent effects emerged, highlighting the importance of matrix-specific evaluation. Conclusions: These findings identify 3 kDa ultrafiltration as an effective and minimally invasive strategy to improve the compatibility of INFOGEST digests with intestinal cell models. By enabling reproducible exposure conditions that preserve epithelial integrity, this approach supports more reliable in vitro assessment of nutrient bioavailability and contributes to methodological standardization in nutrition research. Full article