Search Results (387)

Pigeonpox is a significant infectious disease caused by Pigeonpox virus (PPV), which severely impacts the pigeon industry. Current control methods primarily rely on heterologous vaccines, such as those derived from avian poxviruses, but their protection is limited, creating an urgent need for the development of a specific vaccine. In this study, 720 samples collected from several regions of China between 2022 and 2023 were tested for PPV, followed by virus isolation, identification, and genetic evolutionary analysis. Based on these findings, complete genome sequencing and attenuation of the representative BJ-02 isolate were conducted, and the potential of this strain as a candidate for an attenuated vaccine was preliminarily evaluated. The survey showed PCR positive rates of 9.05%, 16.11%, and 12.50% in samples from Beijing, Guangdong, and Hainan, respectively. Six viral strains were isolated, all of which produced typical lesions on chorioallantoic membranes (CAM) and chicken embryo fibroblasts (CEF). Phylogenetic analysis based on the P4b gene revealed that the six viruses clustered within the same evolutionary branch, closely related to PPV and penguinpox virus strains from South Africa, India, and Taiwan, China. Complete genome sequencing of the BJ-02 strain showed its genomic structure to be similar to that of other fowlpox viruses, with some differences. After serial passage in CAM, PEF and CEF, the BJ-02 SD55 high-passage strain adapted well to in vitro culture, exhibited significantly reduced pathogenicity in chicken embryos and pigeons, and showed no reversion to virulence after five consecutive back-passages. Animal immunization tests demonstrated that the BJ-02 SD55 suspected attenuated strain induced specific antibodies and provided 100% protection against challenge with the virulent strain. In conclusion, PPV is widely prevalent in China. The BJ-02 strain, successfully isolated and attenuated through serial passage, demonstrates excellent immunogenicity and high safety, making it a promising candidate for a specific pigeonpox vaccine. Additionally, the complete genome characterization of BJ-02 contributes to the avian poxvirus genome database and provides critical data to support research on viral pathogenesis and the development of viral vector vaccines for avian and potentially mammalian species. Full article

Background/Objectives: Fungal keratitis remains a vision-threatening infection, and current amphotericin B (AmphB) eye drops suffer from low corneal residence time, poor aqueous solubility, and the need for frequent dosing. This study develops electrospun nanofiber-based ophthalmic inserts combining polyvinyl alcohol (PVA), gamma-cyclodextrin (γ-CD), and sodium taurocholate (STC) to enhance AmphB solubility and provide a non-invasive, rapidly dissolving ophthalmic dosage form. Methods: γ-CD and STC-enhanced AmphB-loaded PVA nanofiber-based ophthalmic inserts with varying γ-CD and STC concentrations were prepared by electrospinning and characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). Drug content, in vitro release (Weibull modeling), antifungal activity against Candida albicans, Fusarium solani, and Aspergillus fumigatus, ocular cytocompatibility using the Hen’s Egg Test on Chorioallantoic Membrane (HET-CAM), and accelerated stability (40 ± 2 °C, 75 ± 5% relative humidity, 4 weeks) were evaluated. Results: Bead-free nanofibers with mean diameters between 216 ± 33 nm and 310 ± 35 nm were obtained, and XRD confirmed complete amorphization of AmphB within the PVA nanofiber matrix, forming an amorphous solid dispersion. All formulations showed rapid and nearly complete AmphB release (≈100% within 60 min), with Weibull β values < 0.75, indicating Fickian diffusion-controlled release. AmphB-loaded PVA nanofiber-based ophthalmic inserts produced inhibition zones and broth susceptibility profiles comparable to AmphB in dimethyl sulfoxide (DMSO), demonstrating preserved antifungal activity. HET-CAM scores (0–0.9) classified the inserts as practically non-irritant, and SEM/FTIR after accelerated storage showed no relevant morphological or physicochemical changes. Conclusions: These γ-CD and STC-enhanced AmphB-loaded PVA nanofiber-based ophthalmic inserts provide a non-invasive, rapidly dissolving ophthalmic dosage form that combines amorphous AmphB, immediate drug availability, and good ocular tolerance, supporting their further development as a patient-friendly treatment option for fungal keratitis. Full article

Background/Objectives: Compounded vaginal creams are widely used for conditions such as hormone replacement therapy, vaginal dryness, low libido, vaginal infections, etc. Recent research highlights the potential of using anhydrous bases to extend shelf life, particularly when combined with self-emulsifying and mucoadhesive properties that improve mucosal retention and enhance drug bioavailability. This study provides in vitro and ex vivo evaluation of an anhydrous vaginal base. Methods: Key quality indicators such as irritation potential, leakage potential, pH compatibility, mucoadhesion, and self-emulsification were assessed using the chorioallantoic membrane Hen’s Egg Test, MTT assay, texture analysis, and fluorescence microscopy. Results: The anhydrous vaginal base demonstrated high cell viability (>78%) and non-irritant potential (IS = 2.5) in in vitro assays. It maintained physiological vaginal pH (4.56 ± 0.05), showed strong mucoadhesive properties comparable to commercial products, and exhibited minimal leakage. Ex vivo studies confirmed its prolonged retention on vaginal tissues. The anhydrous vaginal base formed stable emulsions upon contact with vaginal fluid simulant, effectively distributing both lipophilic and hydrophilic compounds. Conclusions: Compared to water-containing bases, an anhydrous vaginal base shows advantages: longer retention time and lower leakage; adaptability to varying vaginal fluid levels; and efficient dispersion of both hydrophilic and lipophilic active pharmaceutical ingredients. These features support its potential use in compounded vaginal products, minimizing stability risks and enhancing patient compliance and therapeutic outcomes. Full article

Background: Plant-derived essential oils possess valuable bioactivities, but their application is limited by volatility and irritation, which may be addressed through natural polymer encapsulation. This study aimed to investigate the bioactivity of Hedychium coronarium rhizome essential oil and evaluate the effect of microencapsulation on its physicochemical characteristics, biological stability, and irritation profile. Methods: Essential oil was extracted from H. coronarium rhizomes by hydrodistillation and chemically characterized. Enzyme inhibitory activities against elastase, hyaluronidase, and tyrosinase were assessed. Microencapsulation was performed using gum Arabic or maltodextrin at 1–5% w/w oil loadings. The resulting powders were evaluated for morphology, entrapment efficiency, hygroscopicity, water activity, biological stability, and irritation potential using the hen’s egg test on the chorioallantoic membrane. Results: The essential oil demonstrated strong enzyme inhibition, particularly against hyaluronidase (IC₅₀ = 0.1 ± 0.0 µg/mL), along with notable elastase and tyrosinase inhibition. Encapsulation significantly reduced irritation scores from 13.3 ± 1.4 for the free oil to 3.6–4.2 for encapsulated systems (p < 0.05). Gum Arabic produced rough, porous particles with lower hygroscopicity, while maltodextrin yielded smoother particles with lower water activity. Both encapsulated powders significantly enhanced biological stability compared with the ethanolic solution. Conclusions: Natural polymer-based microencapsulation effectively reduced the irritation potential and improved the handling properties of H. coronarium essential oil, supporting its potential application in topical bioactive delivery systems. Full article

Graphene oxide (GO) is a promising nanocarrier for the delivery of oligonucleotides. It offers a high loading capacity, efficient cellular uptake, and surface functionalization. MicroRNA-21 (miR-21) is a well-characterized oncomiR commonly overexpressed in hepatocellular carcinoma (HCC). In HCC, miR-21 contributes to tumor progression, inflammation, and angiogenesis. In a previous in vitro study, we showed that GO alone induces the upregulation of pro-inflammatory and tumor-related genes in HepG2 cells. However, conjugation with an antisense miR-21 (GO-antisense miRNA 21) reverses this effect, suggesting a potential therapeutic application. This study aims to evaluate the antitumor and anti-angiogenic efficacy of the GO-antisense miR-21 nanosystem in ovo using the chick embryo chorioallantoic membrane (CAM) model. Fertilized chicken eggs (n = 4 per group) were randomized into untreated, GO-treated, and GO–antisense miR-21-treated cohorts. A dose of 200 μL (GO 10.0 µg/mL: antisense miR-21 5.0 pmol/mL) was administered intratumorally. Tumor size, volume, and vascularization were monitored through stereomicroscopy and histological analysis. The expression of inflammatory and tumor-associated genes (IL-8, MCP-1, TIMP-2, ICAM-1 and NF-kB) was assessed by quantitative PCR. Given its prominent response, IL-8 protein expression was further analyzed via immunofluorescence. To evaluate tumor-specific delivery, FITC-labeled GO was tracked by confocal microscopy. Our data revealed that treatment with unfunctionalized graphene oxide (GO) unexpectedly promoted tumor vascularization and led to a significant increase in tumor weight. This was accompanied by upregulation of inflammatory markers. In contrast, GO-antisense miR-21 significantly reduced the tumor volume and vessel density. It also successfully downregulated all target genes. Confocal imaging demonstrated preferential accumulation of the nanosystem within the tumor mass. Our results highlight the dual anti-inflammatory and anti-angiogenic effects of GO-antisense miRNA 21 in ovo and support its potential as a targeted nanoplatform for HCC treatment. Full article

Hydrogel-based scaffolds are central to three-dimensional (3D) epithelial culture systems, yet commonly used matrices such as Matrigel^® suffer from batch variability, undefined composition, and limited translational relevance. Here, we comparatively evaluated an animal-free nanocellulose hydrogel (GrowDex^®) and Matrigel^® in a hybrid vascularized intestinal–chorioallantoic membrane (CAM) model. Pre-cultured epithelial–immune constructs (Caco-2/HT29-MTX with immune components) were embedded in both matrices and grafted onto the CAM for 72 h. Histological and immunohistochemical analyses revealed that nanocellulose-based constructs maintained more cohesive epithelial coverage, improved scaffold integrity, and yielded a more continuous cytokeratin-positive layer at the scaffold–CAM interface. In contrast, Matrigel^® constructs frequently exhibited heterogeneous epithelial distribution and central discontinuities. While both matrices enabled CAM engraftment, the chemically defined nanocellulose hydrogel demonstrated enhanced structural robustness during in vivo exposure and histological processing. These findings highlight the suitability of standardized nanocellulose hydrogels for reproducible scaffold-based epithelial models in vascularized environments. Full article

Background/Objectives: Osteosarcoma treatment options remain limited due to tumor metastasis and the toxicity of conventional chemotherapy, warranting new therapeutic strategies. A well-founded strategy is the use of flavonoids, a class of phytochemicals possessing pharmaceutical properties that contribute to anticancer effects, including antioxidant and anti-inflammatory properties. This study aimed to evaluate the anticancer potential of flavonoids in osteosarcoma and investigate their interaction with doxorubicin. Methods: In this study, five flavonoids were screened for cytotoxicity and selectivity across four osteosarcoma cell lines and healthy fibroblasts (MRC-5). The interaction between myricetin and doxorubicin was assessed using a fixed-ratio combination approach. Cell migration and invasion were evaluated using cell exclusion/wound healing and 2D co-culture assays. EMT-related gene expressions were assessed by RT-qPCR. Antitumor activity was evaluated in vivo using a chick chorioallantoic membrane (CAM) xenograft model. Results: Myricetin emerged as the most selective compound, exhibiting cytotoxicity against osteosarcoma cells while sparing MRC-5 fibroblasts. Notably, myricetin synergized with doxorubicin (ratio 69:1), enhancing its cytotoxicity and significantly reducing osteosarcoma cell migration in vitro. Myricetin downregulated SNAI1 and MMP9, suggesting modulation of epithelial–mesenchymal transition (EMT)-related pathways. Complementarily, in the CAM xenograft model, myricetin reduced xenograft tumor size, confirming its anticancer activity in vivo. Conclusions: Collectively, these findings emphasize the anticancer potential of myricetin in osteosarcoma through inhibition of the SNAI1/MMP-9 signaling axis. Full article

Despite advances in periodontal regenerative therapies, consistent tissue regeneration remains challenging, with cells playing an essential role in successful repair. Therefore, this study tested different dental bone substitutes embedded in the chorioallantoic membrane (CAM) combined with periosteum-derived micrografts obtained using a chair-side device (Rigenera HBW system). Cell populations within the micrografts were identified and characterised via immunofluorescence and flow cytometry (CD31, CD105, CD34, CD90, CD73, and CD45). A CAM model was employed to examine the angiogenic potential of micrografts combined with bone substitutes, which were analysed through quantitative blood vessel/vascularisation assessments using the Ikosa software (2025), along with histological and immunohistochemical evaluations such as smooth muscle actin (SMA), H&E, and Masson’s trichrome staining. Statistical analysis was performed using GraphPad Prism 10. The addition of periosteum-derived micrografts resulted in angiogenic enhancement compared to the controls. Notable enhancement of total vessel area, total length, and branching points were obtained when Fisiograft^® (p = 0.0007, p = 0.0002, and p < 0.0001, respectively), New Shore^® (p = 0.0006, p = 0.0149, and p = 0.0083, respectively), and Bio-Oss^® (p = 0.0038 and p = 0.0010, respectively) were combined with micrografts, compared to the positive controls. The histological and immunohistochemical analyses confirmed increased vascularisation (positive staining for SMA) in the micrograft groups. Periosteum-derived micrografts represent a promising adjunct to conventional bone-grafting materials, promoting vascularisation and potentially enhancing tissue regeneration and healing outcomes. Full article

This study evaluated Apis mellifera brood fat extracts as a sustainable alternative to beeswax for anti-inflammatory topical delivery, including their formulation into nanostructured lipid carriers (NLCs). Brood fat was extracted using acetone, ethyl acetate (EA), and hexane, and the resulting extracts were characterized for fatty acid composition and physicochemical properties. Safety was assessed using the hen’s egg chorioallantoic membrane test and cytotoxicity testing in RAW 264.7 macrophages. Anti-inflammatory activity was assessed by inhibition of lipopolysaccharide-induced interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) production. The most suitable extract was formulated into NLCs using sugar squalane as liquid lipid, and the effects of lipid ratio and preparation method were investigated. The results showed that the ethyl acetate extract had the highest yield. Compared with beeswax, all fat extracts exhibited a favorable oleic acid–rich fatty acid profile with comparable crystallinity and thermal behavior, while showing significantly enhanced anti-inflammatory activity (p < 0.05). All extracts and their NLCs were non-irritating and non-cytotoxic. Ethyl acetate extract-based NLCs exhibited favorable particle sizes (72.1 ± 0.3 nm) and narrow polydispersity (0.14 ± 0.00), with high-pressure homogenization producing smaller particles compared to probe sonication without affecting IL-6 or TNF-α inhibition. Therefore, A. mellifera brood fat extract is a sustainable anti-inflammatory lipid source with strong potential as an alternative to beeswax in topical nano-formulations. Full article

Background: Surgical tendon rupture repair suffers from scar formation, leading to tendons with inferior mechanics and consequently to re-ruptures, as well as from adhesion formation to the surrounding tissue, reducing the range of motion. In an approach of re-purposing the phytochemical Bakuchiol to be incorporated in the polymer DegraPol^® (DP), we fabricated a novel implant material by emulsion electrospinning. Methods: To characterize the emulsion electrospun novel materials, we used Scanning Electron Microscopy (SEM) to determine the fiber diameter and pore size. In addition, we used Fourier Transformed Infrared Spectroscopy (FTIR). Finally, we planted the materials onto the chorioallantoic membrane of the chicken embryo (CAM assay) to assess tissue integration and collagen expression. Results: While the pure DP meshes were very well integrated in the CAM assay and showed a significantly higher collagen deposition within the scaffold, the DP + Bakuchiol meshes exhibited poor tissue integration, showing rather the beginning of a fibrous encapsulation. Conclusions: The novel electrospun material DP + Bakuchiol could be used as an anti-adhesion barrier to prevent tendon adhesion. Full article

Circulating tumor cells (CTCs) represent a powerful, minimally invasive window into tumor biology and disease evolution. Technological progress over the past decade has markedly improved the ability to isolate, preserve, and interrogate viable CTCs, transforming them from simple prognostic markers to functional tools for precision oncology. Advances in microfluidic platforms, immunomagnetic enrichment, aptamer-based capture, and nanostructured interfaces have expanded the efficiency and fidelity of CTC recovery, enabling comprehensive molecular profiling and ex vivo analysis. These innovations have paved the way for the development of CTC-derived preclinical models, including xenografts, organoids, and chorioallantoic membrane assays, which recapitulate patient-specific tumor heterogeneity and support individualized drug-sensitivity testing. In this review, we summarize current technologies for CTC isolation, outline recent achievements in functional and pharmacological characterization, and discuss the translational impact of CTC-derived models. We further identify persistent challenges and emerging opportunities, highlighting how integration of multi-omics platforms, artificial intelligence, and standardized workflows may accelerate the clinical implementation of CTC-guided personalized therapy. Full article

Background/Objectives: Insect oils have gained attention as sustainable cosmetic ingredients due to their bioactive lipid content. This study aimed to characterize oils from cricket and to evaluate their safety, biological activities, and performance in sunscreen formulations. Methods: Oils were extracted from Gryllus bimaculatus, Teleogryllus mitratus, and Acheta domesticus by cold pressing following hot-air drying. Fatty acid composition was determined using gas chromatography–mass spectrometry. Safety was assessed by cytotoxicity testing in normal human dermal fibroblasts (NHDF) and the hen’s egg chorioallantoic membrane (HET-CAM) assay. Antioxidant and anti-inflammatory activities were evaluated by intracellular reactive oxygen species (ROS) and nitric oxide (NO^•) assays. Based on biological performance, T. mitratus oil (TMO) was incorporated into sunscreen creams containing physical and chemical ultraviolet (UV) filters. Physical stability, viscosity, pH, sun protection factor (SPF), persistent pigment darkening/ultraviolet A protection factor (PPD/UVA-PF), and blue light protection were evaluated. Results: All cricket oils were non-cytotoxic to NHDF cells and were classified as non-irritating in the HET-CAM assay. TMO exhibited the strongest antioxidant activity, reducing intracellular ROS and significantly inhibiting NO^• production in lipopolysaccharide-stimulated cells. Only TMO showed measurable UVA protection (PPD/UVA-PF = 12.1, PA+++). Sunscreen creams formulated with TMO achieved higher photoprotective efficacy than olive oil-based creams, with SPF values up to 40.51 and PPD/UVA-PF up to 39.17. The inclusion of foundation pigments further increased SPF to 43.09 and enhanced blue light protection to 35.1%. Conclusions: TMO is a safe and effective multifunctional ingredient that enhances sunscreen performance and supports sustainable cosmetic formulation. Full article

Background/Objectives: This study aimed to evaluate the chemical composition and antimicrobial activity of Cordia verbenacea essential oil (EOCV) and to develop and characterize a gel-based nanoemulsion of C. verbenacea essential oil (NECV). Methods: The EOCV was chemically characterized by gas chromatography (GC-FID and GC-MS). The nanoemulsion was prepared using EOCV, the surfactants Cremophor and Plurol Oleique, and phosphate buffer at pH 5.5, and was subjected to experiments to determine its stability, irritant potential and in vitro skin permeation. Results: The main chemical compounds identified in EOCV were α-pinene (33.05%) and β-caryophyllene (25.11%). The EOCV exhibited antimicrobial activity with MIC and MBC values ranging from 6.3 to 25.0 µL/mL for the yeasts Candida albicans and C. krusei, 11.3 to 25.0 µL/mL for the Gram-positive bacteria Bacillus cereus, Staphylococcus aureus, and Enterococcus faecalis, and 12.5 to 75.0 µL/mL for the Gram-negative bacteria Acinetobacter baumannii, Klebsiella pneumoniae, and Escherichia coli. The NECV showed a droplet size of 126.80 nm, a polydispersity index (PDI) of 0.49, a zeta potential of −18.50 mV and a pH of 5.3 and remained stable for 60 days of storage at 25 °C and 4 °C. The HET-CAM irritation test showed that the formulation is non-irritating. The in vitro skin permeation assay showed that the NECV penetrated the deeper layers of the skin, demonstrating its ability to overcome the Stratum corneum barrier. Conclusions: These results are highly promising regarding the potential use of NECV for topical application for the treatment of infected skin wounds. Full article

A novel co-encapsulation platform based on curcumin-loaded liposomes (Cur-Lip) incorporated into thermosensitive hydrogels (TSH) was developed to address the physicochemical and biological limitations of topical curcumin (Cur) delivery. Response Surface Methodology (RSM) was used to optimize Pluronic^® F-127, glycerol, and alginate concentrations with respect to gelation time and viscosity. The optimized formulation (22% Pluronic^® F-127, 5% glycerol, and 0.5% alginate) exhibited rapid time sol–gel transition (~86 s), suitable viscosity (~377 mPa·s), excellent model fitting (R² = 0.99) and prediction accuracy. Three formulations (TSH, Cur-TSH, and Cur-Lip-TSH) were subsequently prepared and displayed appropriate thermoresponsive behavior. The Cur-Lip system showed high encapsulation efficiency (~78%). Upon incorporation into the TSH, Cur-Lip-TSH displayed increased viscosity and mechanical strength at physiological temperature. In vitro studies confirmed its cytocompatibility toward human keratinocytes, significant antibacterial activity against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa, and no irritation potential as assessed by the Hen’s Egg Test on the Chorioallantoic Membrane assay (HET-CAM). Overall, Cur-Lip-TSH represents a safe and robust thermosensitive platform that provides a foundation for future studies on controlled curcumin release and topical performance. Full article

Background/Objectives: Bladder cancer remains a therapeutically challenging malignancy due to high recurrence rates, progression to muscle-invasive disease, and frequent resistance to cisplatin-based chemotherapy. Cold physical plasma (hereafter referred to as plasma) has emerged as a locally applicable modality that generates reactive oxygen species (ROS) and shows preclinical antitumor activity, offering a potential strategy to enhance cisplatin efficacy while enabling dose reduction. Here, we investigated combination treatment with cisplatin and argon plasma generated by the clinically approved kINPen jet in human bladder cancer models. Methods: Three bladder cancer cell lines representing distinct entities were used, namely the urothelial carcinoma lines RT-112 and T24, and the squamous cell carcinoma line SCaBER. IC₂₅ values for plasma and cisplatin monotherapy were established by resazurin assay and used to design combination regimens. Treatment interactions were quantified by coefficient of drug interaction (CDI) analysis and monitored kinetically by long-term live-cell imaging. Plasma-derived ROS were measured in PBS and DMEM, and their functional relevance was assessed in SCaBER cells using catalase and N-acetylcysteine. In ovo validation was performed in the tumor chorioallantoic membrane (TUM-CAM) model, where tumor mass, vascularization, cellular marker expression, and cytokine secretion were analyzed. Results: Plasma and cisplatin exhibited opposing monotherapy sensitivity profiles across cell lines, creating a favorable basis for combination treatment. CDI analysis revealed clear synergy in SCaBER at intermediate cisplatin concentrations, additive effects in RT-112, and additive to mildly synergistic effects in T24. ROS profiling and scavenger experiments identified hydrogen peroxide as a key mediator of plasma and plasma–cisplatin cytotoxicity in SCaBER. In the TUM-CAM model, plasma and cisplatin monotherapies showed notable antitumoral potential. At the same time, plasma–cisplatin combination therapy elicited only modest effects on tumor growth and vascularization compared to monotreatments but induced distinct, cell line-specific alterations in cytokine and marker expression. Conclusions: These findings demonstrate that plasma can potentiate cisplatin cytotoxicity in bladder cancer cells and reshape tumor-associated molecular signatures, supporting further optimization and preclinical evaluation of plasma–cisplatin combination therapy. Full article