Search Results (100)

Cellular senescence is a heterogeneous and dynamic state characterised by stable proliferation arrest, macromolecular damage and metabolic remodelling. Although markers such as SA-β-galactosidase staining, yH2AX foci and p53 activation are widely used as de facto standards, they are imperfect and differ in terms of sensitivity, specificity and dependence on context. We present a multifactorial imaging platform integrating scanning electron, flow cytometry and high-resolution confocal microscopy. This allows us to identify senescence phenotypes in three in vitro models: replicative ageing via serial passaging; dose-graded genotoxic stress under serum deprivation; and primary fibroblasts from young and elderly donors. We present a multimodal imaging framework to characterise senescence-associated phenotypes by integrating LysoTracker and MitoTracker microscopy and SA-β-gal/FACS, p16INK4a immunostaining provides independent confirmation of proliferative arrest. Combined nutrient deprivation and genotoxic challenge elicited the most pronounced and concordant organelle alterations relative to single stressors, aligning with age-donor differences. Our approach integrates structural and functional readouts across modalities, reducing the impact of phenotypic heterogeneity and providing reproducible multiparametric endpoints. Although the framework focuses on a robustly validated panel of phenotypes, it is extensible by nature and sensitive to distributional shifts. This allows both drug-specific redistribution of established markers and the emergence of atypical or transient phenotypes to be detected. This flexibility renders the platform suitable for comparative studies and the screening of senolytics and geroprotectors, as well as for refining the evolving landscape of senescence-associated states. Full article

This study explores an innovative delivery strategy for the management of skin conditions: lipid nanosystems incorporated into a gel matrix. Echinacea purpurea extract, known for its antibacterial, antioxidant, and wound-healing properties, was encapsulated into lipid-based nanosystems and subsequently incorporated into Carbopol-based gel. The extract, rich in chicoric and caftaric acids, exhibited strong antioxidant activity (IC₅₀ = 56.9 µg/mL). The resulting nanosystems showed nanometric size (about 200 nm), high entrapment efficiency (63.10–75.15%), and excellent short-term stability. Superior biocompatibility of the nanosystems, compared to the free extract, was demonstrated using an MTS assay on L-929 fibroblasts. Moreover, the cytoprotective potential of the lipid carriers was evident, as pre-treatment significantly increased cell viability under H₂O₂-induced oxidative stress. These findings suggest that lipid-based encapsulation enhances the therapeutic profile of E. purpurea. The optimal lipid formulation was incorporated into a Carbopol-based gel, which demonstrated an appropriate pH (5.15 ± 0.75), favorable textural properties, sustained polyphenol release, and overall good stability. This research highlights the potential of plant-derived bioactives in the development of dermatocosmetic products, aligning with current trends in eco-conscious and sustainable skincare. Full article

Medicinal plants remain central to traditional healthcare, yet their increasing integration into modern pharmacology necessitates robust toxicological evaluation. Origanum majorana L. (sweet marjoram), widely used in culinary and folk medicine, contains diverse secondary metabolites with both therapeutic and potential genotoxic activities. Despite its popularity, systematic in vivo and in vitro safety assessments remain limited. This study aimed to comprehensively evaluate the acute oral toxicity, cytotoxicity, and genotoxicity of O. majorana methanolic extract, providing baseline toxicological data to support its safe traditional use and potential pharmaceutical applications. The methanol extract of O. majorana leaves was tested in NIH-3T3 fibroblasts for cytotoxicity and genotoxicity. In vivo acute oral toxicity was assessed in rats according to OECD Guideline 420, with animals monitored over 14 days for clinical signs, hematological and biochemical alterations, and histopathological changes. The extract preserved fibroblast viability above 90% across all tested concentrations (10–200 µg/mL), indicating absence of cytotoxicity. However, comet and micronucleus assays revealed dose-dependent DNA damage, suggesting genotoxic potential at higher exposures. In vivo, no mortality or overt systemic toxicity was observed at doses up to 2000 mg/kg. Hematological analyses showed immunomodulatory shifts (increased neutrophils and monocytes, reduced eosinophils), while biochemical profiles indicated hepatoprotective and cardioprotective effects, with reduced ALT, AST, and LDH levels. Histopathological evaluation revealed only mild, focal changes consistent with adaptive rather than irreversible responses. O. majorana extract demonstrates a favorable acute safety profile with preserved hepatic and renal function, hematological modulation, and absence of in vitro cytotoxicity. Nevertheless, dose-dependent genotoxicity warrants caution for concentrated formulations. According to GHS classification, the extract aligns with Category 5 (acute oral toxicity, lowest hazard) and Category 2 (germ cell mutagenicity). These findings underscore the importance of dose management and further long-term genotoxicity studies before translational applications in nutraceutical or biomedical fields. Full article

The use of orthodontic aligners has increased significantly due to their convenience and esthetic advantages. However, understanding their microbiological behavior and cytotoxicity is essential. This study aimed to evaluate the metabolic activity (MA) and proliferation of different bacterial strains—assessed through colony-forming unit (CFU) counts—as well as the cytotoxicity of three widely used aligner systems: Spark, Invisalign, and Smile. Wettability and surface free energy (both dispersive and polar components) were determined using the sessile drop technique. The bacterial strains Streptococcus oralis, Actinomyces viscosus, Streptococcus gordonii, Enterococcus faecalis, and Porphyromonas gingivalis were cultured, and their behavior on the aligner surfaces was assessed under simulated oral cavity conditions in both aerobic and anaerobic environments using a bioreactor. Cytocompatibility was evaluated with HFF-1 human fibroblasts. Distinct strain-specific behaviors were observed. For Spark aligners, the contact angle was 70.5°, Invisalign 80.6°, and Smile 91.2°, and the surface free energy was 60.8, 66.7, and 74. 2 mJ/m², respectively, highlighting the high polar component of the Spark aligner of 31.9 mJ/m² compared to 19.3 and 20.2 mJ/m² for Invisalign and Smile, respectively. The Spark aligner exhibited the lowest metabolic activity for Streptococcus oralis (23.1%), Actinomyces viscosus (43.2%), Porphyromonas gingivalis (17.7%), and biofilm formation (2.4%), likely due to its higher hydrophilicity. The Smile aligner showed the lowest metabolic activity for Streptococcus gordonii (23.6%) and Enterococcus faecalis (51.1%), attributed to its low polar surface free energy component. CFU counts were minimal for all aligners and bacterial strains, including biofilm. All aligners demonstrated cytocompatibility above 70% (Spark: 71.0%, Invisalign: 75.7%, and Smile: 75.6%). These findings highlight the importance of considering aligner material properties in clinical practice and underscore the need for proper oral hygiene and aligner maintenance. Full article

Bixin, an apocarotenoid from Bixa orellana seeds, is a valuable natural pigment with industrial and pharmacological applications. Traditional extraction methods rely on organic solvents, but eco-friendly alternatives like silk fibroin solution (SFS) are emerging. This study evaluated SFS for bixin extraction from annatto seeds, optimizing conditions using Box-Behnken Design (BBD). The optimal parameters 1.5% SFS, 60 °C, and 60 min yielded 10.87 mg/mL (liquid extract of annatto seeds, LEAS + SFS) and 150.72 mg/g (solid extract of annatto seeds, SEAS + SFS). Cell viability was assessed in human dermal fibroblasts (HDFn) and RAW 264.7 murine macrophages via MTT assay. After 24 and 72 h, LEAS + SFS, SEAS + SFS, purified bixin (PB), and SFS maintained >70% viability in HDFn cells. Similarly, RAW 264.7 cells showed >70% viability after 24 h, indicating low cytotoxicity. These results highlight the biocompatibility of SFS-extracted bixin, supporting its potential in food, cosmetics, and biomedicine. The study demonstrates that SFS is an effective, sustainable alternative to traditional solvents, offering high extraction efficiency and minimal toxicity. This method aligns with green chemistry principles, providing a promising solution for bixin production. Full article

This study employs light microscopy, scanning electron microscopy, and transmission electron microscopy to describe the morphological changes occurring during the development of the domestic cat’s uterine horns, originating from the uterine segments of paramesonephric ducts (uPD). Comprehensive observations conducted on 60 specimens aged 28–63 days post-conception (p.c.) revealed that the formation of the endometrium and myometrium in the uterine horns begins around day 33 p.c., initiated by mesenchymal differentiation. During endometrial development, fibroblasts align first in perpendicular and then in oblique columns. The subdivision of the lamina propria into basal and functional layers becomes evident shortly before birth, with the functional layer remaining flat until the end of the prenatal period. The endometrial epithelium transforms from a simple columnar to a pseudostratified structure, undulating by day 63 p.c. Myometrial formation commences with the differentiation of myoblasts, which are arranged in a circular pattern. By the end of gestation, these myoblasts differentiate into smooth muscle cells, organizing into distinct inner circular and outer longitudinal sublayers. Although the fundamental layered architecture of the uterine wall is established before birth, its full maturation—including gland formation, epithelial transformation, and further development of the myometrium—continues postnatally. Full article

This study aimed to evaluate the cytotoxicity of two resin materials, Tera Harz TC-85 DAC and Clear-A, along with the effects of two different post-printing protocols applied to Clear-A. Samples were produced using the Ackuretta Sol printer. The following three groups were formed based on the resins used and the post-curing methods applied: Group 1: Tera Harz TC-85 DAC resin + Tera Harz Cure; Group 2: Clear-A resin + Curie machine; and Group 3: Clear-A resin + Tera Harz Cure. All samples were sterilized in 70% ethanol for 5 min, rinsed with sterile deionized water, and incubated in Dulbecco’s Modified Eagle Medium at 37 °C for 72 h. Cytotoxicity assessment was performed by the XTT and RTCA methods using the human gingival fibroblast cell line. According to the XTT assay, undiluted resin extracts exhibited approximately 75–80% cell viability at 24 h, while further dilutions resulted in a viability exceeding 90%. No significant differences in viability were observed among the groups at any dilution at 48 and 72 h. The xCELLigence RTCA results aligned with the XTT findings, showing a transient decrease in cell viability within the first 24 h, followed by continued cell growth. This study demonstrated that extracts from all tested 3D-printed resins exhibited biocompatibility with human gingival fibroblasts. These findings support their potential for further applications in the dental and biomedical fields. Full article

Wound healing is a complex biological process that benefits from advanced biomaterials capable of modulating inflammation and promoting tissue regeneration. In this study, cerium oxide nanoparticles (CeO₂NPs) were green-synthesized using Hemerocallis citrina extract, which served as both a reducing and stabilizing agent. The CeO₂NPs exhibited a spherical morphology, a face-centered cubic crystalline structure, and an average size of 9.39 nm, as confirmed by UV-Vis spectroscopy, FTIR, XRD, and TEM analyses. These nanoparticles demonstrated no cytotoxicity and promoted fibroblast migration, while significantly suppressing the production of inflammatory mediators (TNF-α, IL-6, iNOS, NO, and ROS) in LPS-stimulated RAW264.7 macrophages. Gene expression analysis indicated M2 macrophage polarization, with upregulation of Arg-1, IL-10, IL-4, and TGF-β. Aligned polycaprolactone/polylactic acid (PCL/PLA) nanofibers embedded with CeO₂NPs were fabricated using electrospinning. The composite nanofibers exhibited desirable physicochemical properties, including porosity, mechanical strength, swelling behavior, and sustained cerium ions release. In a rat full-thickness wound model, the CeO₂ nanofiber-treated group showed a 22% enhancement in wound closure compared to the control on day 11. Histological evaluation revealed reduced inflammation, enhanced granulation tissue, neovascularization, and increased collagen deposition. These results highlight the therapeutic potential of CeO₂-incorporated nanofiber scaffolds for accelerated wound repair and inflammation modulation. Full article

The extracellular matrix (ECM) provides structural support to cells, thereby forming a functional tissue. In cancer, the growth of the tumor creates internal mechanical stress, which, together with the remodeling activity of tumor cells and fibroblasts, alters the ECM structure, leading to an increased stiffness of the pathological ECM. The enhanced ECM stiffness, in turn, stimulates tumor growth and activates tumor-promoting fibroblasts and tumor cell migration, leading to metastasis and increased therapy resistance. While the relationship between matrix stiffness and migration has been studied before, their connection to internal tumor stress remains unresolved. Here we used 3D ECM-embedded spheroids and hydrogel particle stress sensors to quantify and correlate internal tumor spheroid pressure, ECM stiffness, ECM remodeling, and tumor cell migration. We note that 4T1 breast cancer spheroids and SV80 fibroblast spheroids showed increased invasion—described by area, complexity, number of branches, and branch area—in a stiffer, cross-linked ECM. On the other hand, changing ECM stiffness only minimally changed the radial alignment of fibers but highly changed the amount of fibers. For both cell types, the pressure measured in spheroids gradually decreased as the distance into the ECM increased. For 4T1 spheroids, increased ECM stiffness resulted in a further reach of mechanical stress into the ECM, which, together with the invasive phenotype, was reduced by inhibition of ROCK-mediated contractility. By contrast, such correlation between ECM stiffness and stress-reach was not observed for SV80 spheroids. Our findings connect ECM stiffness with tumor invasion, ECM remodeling, and the reach of tumor-induced mechanical stress into the ECM. Such mechanical connections between tumor and ECM are expected to drive early steps in cancer metastasis. Full article

Limnospira spp., commercially known as spirulina, is widely recognized for its remarkable benefits due to its rich composition of bioactive compounds like phycobiliproteins, carotenoids, and phenolic compounds. These natural bioactive compounds not only serve as colorants but also offer potent antioxidant, anti-inflammatory, immunomodulatory, anticancer, antimicrobial, and anti-aging properties. As a result, spirulina and its components are increasingly used in cosmetic formulations to promote skin hydration, reduce wrinkles, and protect against UV radiation damage. Its bioactive components enhance fibroblast growth, boost collagen production, and prevent premature skin aging by inhibiting enzymes responsible for elastin degradation. Additionally, spirulina-based cosmetics have demonstrated wound-healing properties without genotoxic effects, with formulations containing C-phycocyanin particularly effective in shielding skin cells from UV-induced apoptosis. Despite these well-established benefits, there remains significant potential for the cosmetic industry to harness spirulina’s capabilities further. Research into the molecular mechanisms underlying its bioactive compounds in cosmetic formulations is still in its early stages, offering many opportunities for innovation. Emerging fields of biotechnology, such as nanotechnology and biocosmetics, could enhance the stability, efficacy, and delivery of spirulina-based ingredients, unlocking new possibilities for skin protection and rejuvenation. Furthermore, its proven biological properties align perfectly with the increasing consumer demand for safe, sustainable, and nature-inspired skincare solutions. Full article

Although a myocardial infarction occurs roughly every minute in the U.S. alone, medical research has yet to unlock the key to fully enabling post-hypoxic myocardial regeneration. Thymosin beta-4 (TB4), a short, secreted peptide, was shown to possess a beneficial impact regarding myocardial cell survival, coronary re-growth and progenitor cell activation following myocardial infarction in adult mammals. It equally reduces scarring, however, the precise mechanisms through which the peptide assists this phenomenon have not been properly elucidated. Accordingly, the primary aim of our study was to identify novel molecular contributors responsible for the positive impact of TB4 during the remodeling processes of the infarcted heart. We performed miRNA profiling on adult mice hearts following permanent coronary ligation with or without systemic TB4 injection and searched for targets and novel mechanisms through which TB4 may mitigate pathological scarring in the heart. Our results revealed a significant increase in miR139-5p expression and identified ROCK1 as a potential target protein aligned. Real-time PCR, Western blot and immunostaining on adult mouse hearts and human cardiac cells revealed the peptide indirectly or directly modulates ROCK1 protein levels both in vivo and in vitro. We equally discovered TB4 may reverse or inhibit fibroblast/myofibroblast transformation and the potential downstream mechanisms by which TB4 alters cellular responses through ROCK1 are cell type specific. Given the beneficial effects of ROCK1 inhibition in various cardiac pathologies, we propose a potential utilization for TB4 as a ROCK1 inhibitor in the future. Full article

Objective: This study aimed to establish a reproducible protocol for obtaining four plasma fractions (autologous platelet-rich plasma (C-PRP, PRP LCC) and platelet-rich fibrin (I-PRF, F-PRF)) from a single blood draw and to evaluate their effectiveness in enhancing skin density and thickness in facial aesthetic treatments across different age groups. Methodology: Twenty participants aged 30–60 years received three treatments at 4–6 week intervals, with C-PRP, PRP LCC, I-PRF, and F-PRF injections in targeted facial areas. Ultrasound measurements provided an objective method to assess the outcomes, and statistical analysis evaluated the changes in skin density and thickness. A comprehensive literature review contextualized our findings within the broader scientific discourse on PRP and PRF applications in aesthetic medicine. Results: The protocol successfully yielded four distinct plasma fractions from a single blood draw. Ultrasound and statistical analyses demonstrated significant improvements in skin density and thickness, particularly in the lower eyelid area. These improvements were consistent across all the age groups, suggesting that the therapy’s effectiveness is independent of age. The findings also align with previous research, underscoring PRP’s and PRF’s roles in stimulating fibroblast activity, promoting collagen synthesis, and enhancing skin quality. Full article

Background: Nitric oxide (NO) has been linked to the pathogenesis of asbestos-related pleural diseases, including an extremely aggressive cancer called malignant pleural mesothelioma (MPM). Given that MPM cells are characterized by a higher expression of NO synthases and elevated NO production relative to normal cells, the use of NO-donor compounds could potentially saturate the cancerous cells with NO, triggering their death. Methods: We developed a novel class of NO prodrugs by merging two NO-releasing components, 1,2,5-oxadiazole 2-oxides (furoxans) and 1,2,4-oxadiazoles, and studied their NO-releasing characteristics in a time-dependent manner using the Griess assay. The cytotoxicity against two human MPM cell lines and non-cancerous lung fibroblasts was evaluated using a colorimetric MTT assay. Results: All compounds exhibited excellent NO-donating properties, surpassing the capacity of two reference NO donor compounds, 3-carbamoyl-4-(hydroxymethyl)furoxan (CAS-1609) and 4-ethoxy-3-phenylsulphonylfuroxan (CHF-2363), by at least 1.5–3 times. All oxadiazole hybrids demonstrated high cytotoxicity against MPM cell lines in a low micromolar range, comparable or higher than the cytotoxicity of the standard-of-care drug cisplatin. Conclusions: Notably, the novel compounds displayed a markedly greater selectivity towards cancerous cells than cisplatin when compared with non-cancerous lung fibroblasts, aligning with the intended design. Full article

Corneal fibroblasts are central to normal and abnormal wound healing in the cornea. During the wound healing process, several biochemical and biophysical signals that are present in the extracellular matrix (ECM) play critical roles in regulating corneal fibroblast behavior. The translocation and activation of Yes-associated protein (YAP)—a main transcriptional factor in the Hippo signaling pathway—is one example of mechanotransduction involving these signals. However, how corneal fibroblasts integrate these simultaneous cues is unknown. In this study, we utilized well-defined micropatterns of aligned collagen fibrils and other ECM proteins to explore the effects of cell density, topography, geometric confinement, and ECM composition on the translocation of YAP in corneal fibroblasts. We observed that when human corneal fibroblasts (HTKs) were confined to narrow micropatterns (50 μm and 100 μm) of proteins, there was a high degree of cell alignment irrespective of cell seeding density. However, the location of YAP was dependent upon the cell seeding density, ECM composition, and topography. YAP was more nuclear-localized on substrates coated with aligned collagen fibrils or fibronectin as compared to substrates coated with monomeric collagen, random collagen fibrils, or poly-L-Lysine. In addition, we also observed that YAP nuclear localization was significantly reduced when HTKs were cultured on aligned collagen fibrils, monomeric collagen, or fibronectin in the presence of monoclonal blocking antibodies against α₅ or β₁ integrin subunits. Finally, we observed that HTK cells formed fibrillar fibronectin on both monomeric collagen and aligned collagen fibrils. These findings provide new insights into how simultaneous biochemical and biophysical cues affect YAP localization in corneal fibroblasts. Full article

The adhesion and alignment of osteoblasts and fibroblasts on titanium alloy (Ti-6Al-4V) surfaces can be adjusted over a wide range by femtosecond laser treatment and anodization. The great differences in cell behavior between different experimental conditions raised further questions about the role of cell migration, which will be addressed in this study. For that, Ti-6Al-4V surfaces were laser-structured to obtain a surface covered with ripples, i.e., laser-induced periodic surface structures (LIPSS), or micro-cones superimposed with ripples. Then, cells were seeded either directly onto the non-structured or laser-structured areas on the titanium alloy samples or beside such samples where they can reach the surface by cell migration. After two weeks in culture, the cell coverage of the samples was evaluated by scanning electron microscopy (SEM). The results showed that cells directly seeded onto the non-structured or laser-structured areas covered the surface nearly completely and eventually aligned along the ripple direction for the laser-structured areas. In contrast, for cell-seeding beside the samples, the laser-structured areas remain nearly cell-free while the non-structured areas were covered with cells in a similar non-oriented manner as for direct cell-seeding. These results on reduced osteoblast migration due to laser structuring are in line with the findings in animal experiments. There, the new bone formation of laser-processed samples was 26.1% ± 16.9% lower in comparison to untreated samples of the same type, which can be explained by hindered cell migration on the laser-processed areas of the screws. Full article