Search Results (240)

Background: Effective vaccines are essential to overcome the limitations of livestock immunisation, particularly in low- and middle-income countries (LMICs), where scalable, thermostable, and easy-to-administer solutions are needed. Nanoparticle-based delivery systems, such as the Spontaneous Nanoliposome Antigen Particle (SNAP) technology using CoPoP liposomes, offer a promising alternative for subunit vaccine development, although their performance in large animal species remains poorly characterised. CoPoP enables the rapid non-covalent multimeric display of His-tagged protein antigens combined with immunomodulators on liposomes incorporating cobalt porphyrin–phospholipid (CoPoP). Objective: To evaluate the immunogenicity of CoPoP-based liposomes delivering the Theileria parva p67C antigen in cattle and compare their performance in murine models. Methods: Cattle and mice were immunised with p67C formulated in CoPoP liposomes incorporating QS-21 and/or PHAD immunomodulators. Humoral and cellular responses were assessed. Parallel in vitro stimulation of bovine PBMC with Quil-A was used to investigate the mechanistic effects of saponins on bovine cells. Results: CoPoP liposome formulations did not improve p67C immunogenicity in cattle, with antibody responses at least two-fold lower than previously reported results and no detectable cellular responses. In contrast, the same platform induced up to 2000-fold higher antibody titres in mice. This disparity is likely driven by differences in antigen dose relative to body mass, tissue architecture, lymphatic accessibility, and innate immune signalling differences. PHAD-mediated TLR4 activation appeared less effective in cattle, whereas QS-21 induced a broader immune activation, likely through conserved inflammasome pathways. Despite limited immunogenicity, antigen presentation by CoPoP liposomes was preserved. Conclusions: SNAP-based CoPoP liposomes showed strong immunogenicity in mice but limited efficacy in cattle, highlighting the challenges of cross-species translation. Optimisation of antigen dose and adjuvant selection for the targeted species is required, with QS-21 representing a more promising candidate than the TLR4 agonist. The scalability and versatility of SNAP technology support its continued development for multivalent livestock vaccines. Full article

Kiwifruit soft rot is a major cause of postharvest loss owing to rapid fruit decay during storage. This study focused on kiwifruit soft rot during the postharvest storage stage, when fungal development may be promoted by room temperature and high humidity. Soft rot symptoms were observed in the pericarp and fruit flesh. In this study, carvacrol-loaded nanoliposomes (CAR@NL) were prepared by an O/W emulsification–solvent evaporation method to control kiwifruit soft rot. The physicochemical properties of CAR@NL were characterized by laser particle size analysis, Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). Their antifungal activity and preservation efficacy were evaluated by in vitro antifungal assays and fruit storage experiments. The prepared CAR@NL showed an average particle size of approximately 280 nm, an encapsulation efficiency of 85.75%, and a drug loading capacity of 20.14%, along with favorable sustained-release properties. CAR@NL exhibited strong antifungal activity, with an EC₅₀ value of 41.76 mg/L. DAPI staining indicated no obvious effect on fungal DNA, whereas propidium iodide (PI) staining revealed increased fluorescence intensity with increasing concentration and treatment time, indicating disruption of hyphal membrane integrity and severe structural damage. Flow cytometric analysis further showed that, at 50 mg/L, the total apoptosis rate was 2.96% in the untreated control group, 5.22% in the CAR@NL-treated group, and 33.6% in the carbendazim-treated group, demonstrating the lower cytotoxicity of CAR@NL toward mammalian cells. In addition, CAR@NL showed good stability and preservation performance during fruit storage. Overall, CAR@NL may serve as a safe and effective postharvest agent for the control of kiwifruit soft rot. Full article

Background/Objectives: This study addresses the need for scalable and predictive strategies linking mixing conditions to nanocarrier properties by developing and analyzing a coaxial jet antisolvent process for the continuous production of pharmaceutical nanocarriers. Methods: A single experimental platform was used to generate both curcumin-based nanoparticles and nanoliposomes, enabling direct comparison of how mixing regime and formulation variables influence product characteristics. Results: Fluid-dynamic behavior was first characterized using tracer and micromixing experiments, revealing a strong dependence of mixing time on flow conditions, with characteristic mixing times decreasing from >1000 ms under laminar conditions to approximately 10–30 ms in turbulent regimes. Nanoparticles and liposomes obtained under optimized conditions exhibited mean sizes in the range of 120–250 nm, with polydispersity indices typically below 0.2 under optimized turbulent conditions. To rationalize these observations, a computational framework was implemented, combining Reynolds-averaged computational fluid dynamics with a population balance formulation solved by the method of moments. The model provided spatially resolved insight into solvent exchange, supersaturation development, and nucleation–growth dynamics, showing good agreement with experimental trends and capturing the effect of mixing conditions on particle size across different regimes. Conclusions: Although simplified, the modeling approach establishes the basis for future extensions toward full population-balance distribution simulations capable of predicting complete particle size distributions, highlighting the ability of the coaxial jet mixer to control supersaturation and particle formation through tunable hydrodynamic conditions. This capability makes the system particularly attractive compared to conventional batch or less controllable mixing technologies, enabling a more rational and scalable design of pharmaceutical nanocarriers, with good encapsulation performance as discussed in the main text. Full article

Background: Effective anti-aging requires dual strategies to stimulate regeneration and counteract damage. While the combination of hydroxypinacolone retinoate (HPR) and carnosine (CA) holds great promise, their effectiveness is hampered by instability and poor skin penetration. Methods: To overcome these challenges, this study developed HPR and CA co-encapsulated nanoliposomes (HC-NLPs) via high-pressure homogenization as an advanced epidermal/dermal delivery system. Results: HC-NLPs markedly improved skin retention of HPR (58.97%) and CA (111.36%) compared to the free combination (Free-HC). In cellular studies, HC-NLPs displayed excellent biocompatibility and demonstrated a 4.7-fold higher cellular uptake. This led to enhanced proliferative (EdU positive rate increased by 78.32%) and migratory (wound closure improved by 31.5%) capacities. Moreover, HC-NLPs effectively reinforced multiple skin-protective processes associated with aging, including enhanced resistance to oxidative and glycation-induced damage, suppressed inflammatory responses, and strengthened cellular barrier integrity. In 3D skin models, HC-NLPs promoted collagen deposition and improved tissue morphology compared to Free-HC. Their superior in vivo antioxidant and anti-aging effects were further validated in Zebrafish assays. HC-NLPs effectively co-deliver HPR and CA, markedly improving their stability, skin penetration, and cellular internalization. Conclusions: The formulation demonstrates comprehensive pro-regenerative, anti-inflammatory, antioxidative, and anti-glycation effects, representing a promising nano-delivery strategy for advanced anti-aging skincare. Full article

Background/Objectives: Nerolidol (NER) is a sesquiterpene alcohol with recognized antimicrobial potential, whose applications as a pure substance are limited by hydrophobicity, instability, and cytotoxicity. Invasomes, i.e., liposomes with terpene ingredients, offer a strategy to improve their delivery; however, the NER loading limits compatible with vesicle integrity are still unclear. Here, Nerolidol-loaded invasomes were produced using a controlled simil-microfluidic coaxial injection process. Methods and Results: As a preliminary step, unloaded liposomes were fabricated to consolidate operating conditions and ensure their reproducible colloidal properties. Thereafter, formulations with progressively decreasing nominal NER loads were investigated to evaluate vesicle size, polydispersity, ζ-potential, encapsulation efficiency, effective loading, and stability. High nominal loads promoted turbidity, size increase (by agglomeration coalescence phenomena), and structural instability, whereas formulations containing approximately 1–2% NER achieved nearly complete encapsulation, Z-average ≈ 300 nm, |ζ| > 30 mV, and satisfactory physical stability. Antimicrobial and cytotoxic profiles of representative formulations, previously evaluated in an independent study are here reported only to contextualize the practical relevance of the optimized systems, while the present work primarily focuses on process–formulation aspects and loading/stability limitations. Conclusions: Overall, the present work identifies a realistic loading window for Nerolidol invasomes and highlights the suitability of the simil-microfluidic approach to obtain scalable, well-controlled formulations, providing a rational basis for their future biological assessment. Nerolidol invasome systems indeed can be considered a promising, versatile platform for antimicrobial applications, including prospective use in animal feed. Full article

Epigallocatechin gallate (EGCG) represents the key phenolic compound in green tea, which has been verified to possess various biological effects but suffers from low stability and poor bioavailability. To address these issues, EGCG-loaded nanoliposomes (ELs) were screened and prepared using an ethanol injection–calcium acetate gradient (EtOH-CAG) method. An encapsulation efficiency of 94.61% was achieved, involving a particle size of 118.6 nm and a polydispersity index (PDI) of 0.23. Via layer-by-layer assembly, nanoliposomes modified with either ε-polylysine (ε-PL) monolayer (ELP) or ε-polylysine/sodium alginate (SA) bilayer (ELPA) exhibited substantially improved stability. Moreover, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and thermal analysis confirmed the effective loading of EGCG and the successful decoration of ε-PL and SA. Molecular docking analyses of dual ligands further characterized the surface modification mechanism, which was primarily mediated by hydrogen bonding and electrostatic interactions. ELPA maintained robust stability under conditions including 200 mM salt concentration, a pH range of 4–10, temperatures up to 55 °C, and a 25-day storage duration. The modified systems showed considerably enhanced cellular uptake without causing cytotoxicity. Collectively, the developed ε-PL/SA bilayer nanoliposomes offer an eco-friendly, efficient strategy to enhance EGCG stability and in vitro bioavailability in functional food applications. Full article

Background: Prostate cancer is a leading malignancy among males, and conventional chemotherapy is often limited by insufficient tumor selectivity and systemic toxicity. Prostate-specific membrane antigen (PSMA), which is highly expressed on prostate cancer cells, represents a promising target for precision drug delivery. In this study, we developed a folate-modified, PSMA-targeting nanoliposome loaded with docetaxel (DFL) to enhance tumor specificity and therapeutic efficacy. Methods: DFL was prepared using a thin-film hydration–sonication method and characterized through physicochemical analyses. Cellular uptake and cytotoxicity were evaluated in PSMA-high LNCaP cells, with PSMA knockdown used to assess target-dependent internalization. Antitumor efficacy was examined with a microfluidic system and LNCaP xenograft nude mice, and safety was evaluated by measuring hepatic and renal biomarkers and performing histopathological analysis of major organs. Results: DFL demonstrated favorable physicochemical properties and significantly enhanced cellular uptake and cytotoxicity in LNCaP cells relative to control formulations. PSMA knockdown markedly attenuated cellular sensitivity to DFL, confirming PSMA-dependent internalization. A 3D microfluidic perfusion platform further corroborated robust and selective DFL uptake under dynamic flow conditions, thereby strengthening the translational relevance of the targeting effect beyond static cultures. In vivo, DFL substantially inhibited tumor progression in LNCaP xenograft models, reducing both tumor volume and weight by more than 50%. TUNEL assays showed increased apoptosis, and immunohistochemistry revealed reduced Ki-67 expression with concomitant upregulation of Caspase-3. No significant alterations in hepatic or renal biomarkers were observed, and histopathological evaluation demonstrated no treatment-associated lesions in major organs. Conclusions: A folate-modified, PSMA-targeting docetaxel nanoliposome was successfully developed, demonstrating enhanced tumor-specific drug delivery and improved antitumor activity with favorable biocompatibility in preclinical models. DFL represents a promising nanomedicine strategy for the precision chemotherapy of prostate cancer. Full article

Background/Objectives: Gramicidin S (GS) is a cyclic antimicrobial peptide with strong antibacterial activity but significant cytotoxicity toward mammalian cells. This study evaluated GS-induced cytotoxicity in L929 fibroblast cells using both traditional 2D monolayer cultures and more physiologically relevant 3D spheroid models, and assessed whether liposomal encapsulation could mitigate toxicity and improve biocompatibility. Methods: L929 cells were cultured in monolayers and spheroids and treated with free GS or GS encapsulated in liposomes of varying lipid compositions. Cell viability and morphology were evaluated after 24 h of exposure using standard cytotoxicity assays. Results: Control liposomes, regardless of tested lipid type or concentration, showed no adverse effects on cell morphology or viability. Free GS caused pronounced, dose-dependent cytotoxicity in monolayers, decreasing viability to 11.0 ± 1.9% and 0.5 ± 1.1% at 50 and 75 µg/mL, respectively. By contrast, encapsulation in liposomes significantly reduced toxicity (p < 0.05), preserving 80.3–82.2% viability at 75 µg/mL depending on formulation, corresponding to protection factors exceeding 160-fold (80.3% vs. 0.5%). Spheroid cultures showed slightly higher resistance to GS; free GS reduced viability to 2.9%, while liposomal GS preserved it above 84.8%, depending on lipid composition. Conclusions: Liposomal encapsulation effectively reduces GS-induced cytotoxicity, likely by limiting direct membrane disruption. Moreover, spheroid models provide a more physiologically relevant and predictive platform for toxicity testing, while the results support nanoliposomes as a practical delivery strategy to enhance the safety of antimicrobial peptides during preclinical development. Full article

Background/Objectives: Knee osteoarthritis stands as the highest prevalent joint disease worldwide, affecting millions of adults and significantly impairing mobility and quality of life. Pro-inflammatory cells and cytokines are considered key players in the pathophysiology of the disease. In previous work, two anti-inflammatory therapeutic approaches were developed: a secretome enriched in anti-inflammatory cytokines, and nanoliposome-bound TRAIL (LUV-TRAIL), with proven efficacy against rheumatoid arthritis in rabbits. Methods: In this work, we evaluated the ability of these treatments to prevent the development of osteoarthritis (OA) in an ovine model following meniscectomies. Two weeks after the surgeries, knees were treated with several rounds of single or combined therapy, and then sheep were left untreated for several months. Knee damage was followed by X-ray analysis and, after sacrifice, assessed through macroscopic inspection, histological determinations, and inflammatory cytokine measurements. Results: The combined therapy had a significant positive effect against osteoarthritis development. Specifically, the combination is capable of improving knee injury in the first stages of OA in several parameters, such as synovial hyperplasia and tibial plateau damage, which are two of the most frequently damaged areas. Other markers, such as synovial inflammation and X-ray and macroscopic images, also presented a tendency to improved scores. Conclusions: The combination of the secretome with LUV-TRAIL represents a promising therapy worth exploring further in osteoarthritis treatment and/or prevention. Full article

Plant-derived biostimulants represent an innovative approach to enhancing crop productivity, resilience, and quality within sustainable agricultural systems by improving nutrient uptake, stress tolerance, and plant defense mechanisms while reducing reliance on synthetic inputs. However, their effectiveness is often limited by poor stability and low bioavailability. Recent advances in nanotechnology, particularly liposomal formulations, address these limitations by enhancing the stability, solubility, and delivery efficiency of bioactive plant compounds. Liposomes facilitate the penetration and systemic transport of active ingredients within plant tissues and enable controlled release at the target site, thereby increasing biostimulant efficacy. This review summarizes current knowledge on plant-derived biostimulants, their classification, nano-formulation, molecular mechanisms, and roles in mitigating abiotic and biotic stress. Special emphasis is placed on liposome-based formulations, including supercritical CO₂ extracts and nano-liposomal delivery systems, with examples such as garlic extract and the EliceVakcina^® complex. Finally, the potential of liposomal technologies in integrated crop protection and sustainable agriculture is discussed. Full article

Background/Objectives: Nickel exposure is a significant environmental and occupational risk factor associated with the onset and progression of chronic liver diseases due to its capacity to induce persistent oxidative stress, inflammation, and hepatocellular injury. This study aimed to evaluate the enhanced hepatoprotective and antioxidant/anti-inflammatory effects of naringin-loaded nanoliposomes (NRG-NLPs), a novel nanoformulation designed to improve the bioavailability of naringin, a citrus-derived flavonoid phytochemical, against nickel sulfate (NiSO₄)-induced hepatotoxicity in male Wistar rats. Methods: Ninety rats were allocated into six groups (n = 15 each): control, NRG, NRG-NLPs, NiSO₄, NiSO₄ + NRG, and NiSO₄ + NRG-NLPs. Treatments consisted of oral administration of NRG or NRG-NLPs (80 mg/kg/day) and intraperitoneal injections of NiSO₄ (20 mg/kg/day) for three weeks. Endpoints included assessment of growth performance, serum biochemistry, hepatic antioxidant status, inflammatory mediators, apoptotic gene expression, nickel tissue accumulation, and histopathological and ultrastructural liver changes. Results: NiSO₄ exposure induced marked hepatic injury, evidenced by reduced body weight, adverse serum biochemical profiles, increased hepatic enzymes and bilirubin, elevated oxidative damage markers (MDA, protein carbonyls), increased proinflammatory cytokines, and upregulation of HMGB1, PI3K, mTOR, JAK/STAT, and proapoptotic genes, accompanied by aberrant nickel accumulation and severe histopathological alterations. Co-treatment with NRG-NLPs significantly ameliorated biochemical and histological disturbances, restored antioxidant defense systems (SOD, CAT, GPx, GSH, Nrf2, HO-1), and modulated key pathways of inflammation (NF-κB, TNF-α, IL-6), fibrosis (TGF-β), cell survival, and apoptosis more effectively than crude naringin. NRG-NLPs also substantially reduced hepatic nickel deposition and preserved near-normal liver architecture. Conclusions: These findings demonstrate that nanoformulated naringin confers superior hepatoprotective benefits against nickel-induced liver injury through enhanced bioavailability and multi-pathway modulation, supporting its translational potential as a citrus-derived medicinal phytochemical and dietary bioactive for the prevention and therapeutic intervention of oxidative and inflammatory chronic liver disease. Full article

In this study, we aimed to investigate the potential of utilizing red grape pomace as a source of polyphenolic compounds in the growing, fragmented winemaking sector in Poland. For polyphenol extraction, we compared two methods: conventional extraction using water and alcohol solutions, and the supercritical CO₂ technique with ethanol as a cosolvent. The conventional method yielded at least 30% more polyphenols compared to the advanced SC-CO₂ technique. Experimentally chosen conditions, including a solvent composition of ethanol–water (1:1; v/v) containing 3% HCl, a liquid-to-solid ratio of 25:1 mL/g, and 2 min of ultrasound pretreatment and conventional extraction at a temperature of 30 °C over 4.5 h, enabled an extraction efficiency of 101 mg of total polyphenols per 1 g of raw material used, with an antioxidant capacity equivalent to 600 µmol of Trolox. According to HPLC analyses, the main components of the investigated biomass were epicatechin, anthocyanins and p-coumaric acid. The extract was encapsulated in liposomes, revealing no negative effect on their stability or aggregation under the conditions tested (21 days). The study suggests that conventional water–ethanol extraction can be a relatively safe and effective method for managing winemaking residuals, increasing the competitiveness of small producers through the production of high-value antioxidant additives. Full article

This study emphasizes overcoming the challenges of targeted drug delivery in colon cancer therapy by developing gastrointestinal (GI) stable, pectin-coated nanoliposomes for the oral delivery of Apigenin-Cyclodextrin Complex as an in situ gel formation. Initially, the formulation was strategically designed using design expert software for formulation optimization. FTIR and XRD studies were conducted to ensure physical compatibility and to confirm the encapsulation of apigenin within the formulation. In process optimization, among all seventeen formulations run tested, PNL (Api-Cy)-13 was identified for the highest drug loading, favourable size dimension of particle with zeta potential, and spherical external morphology through SEM analysis. The metered drug release during an in vitro study for PNL (Api-Cy)-13 was remarkably high (more than 75% of drug availability in the colonic environment, precisely in contrast to only 20% in the gastric phase in a sustained release manner), focused on colon drug targeting as an in situ gel. Furthermore, apigenin release from PNL (Api-Cy)-13 in an ex vivo chick ileum permeability study was observed both in the absence and presence of 1% vancomycin. An incremental apigenin release in the absence of the antibiotic (1% vancomycin) indicated gut microbial-associated and pectinase-mediated drug release. Here, pectin degradation materializes by the colonic microbial environment, which facilitates desirable incremental colonic drug permeation. Finally, an in vitro MTT assay and a competitive flowcytometric cell uptake study with PNL (Api-Cy)-13 using HCT-116 cells proved significant superiority in cytotoxicity profile for apigenin when delivered as an optimized coated nanoliposome in comparison to free apigenin or other non-modified nano-formulation. Also, the inhibition of the cell efflux process was validated by Multidrug Resistance 1 (MDR1) gene regulation. These observations establish an undoubted promise for the novel biopolymer, pectin-based apigenin-cyclodextrin nanoliposomes as targeted therapy in colon cancer with significant in vivo pharmacokinetics and safety profile. Full article

In the development of functional foods with therapeutic value, nanoliposomal carriers offer a promising strategy for enhancing the stability and efficacy of bioactive compounds in dairy matrices. This study evaluated the sensory acceptance and physicochemical stability of yogurt enriched with anthocyanin-loaded nanoliposomes during 21 days of refrigerated storage, assessing the impact of nanoencapsulation on compound preservation and quality. Nanoliposomes were synthesized using ultrasonic film dispersion and characterized for antioxidant and erythroprotective activities. Antioxidant capacity was assessed through DPPH, ABTS, and FRAP assays, while erythroprotective effects were evaluated via oxidative hemolysis using human erythrocytes of different ABO/RhD phenotypes. These were incorporated into artisanal yogurt, followed by physicochemical, microbiological, rheological, and sensory analyses. Anthocyanins showed strong antioxidant capacity, especially in ABTS (93.24%), DPPH (21.34%), and FRAP (1023.24 µM TE/g D.W.), reflecting their radical scavenging and reducing power. They also exhibited high erythroprotective activity, with greater antihemolytic effects in O RhD− blood and enhanced photoprotection against UVA in O RhD+ blood. Yogurt enriched with nanoliposomes showed improved color stability, reduced syneresis, and favorable rheological and sensory characteristics. These findings support nanoliposomes as molecular delivery systems in functional dairy matrices with potential nutraceutical applications targeting oxidative stress. Further work should explore molecular mechanisms and validate health-promoting effects. Full article

The growing threat of multidrug-resistant fungal pathogens, especially Candida auris, has underscored the need for effective antifungal vaccines. This commentary highlights recent advances in peptide-based vaccination using the SNAP (Spontaneous Nanoliposome Antigen Presentation) platform, focusing on the FM-SNAP vaccine, a bivalent liposomal formulation targeting the surface-expressed peptides fructose bisphosphate aldolase (Fba) and methionine synthase (Met6). Compared to earlier constructs such as MP12, FM-SNAP achieves superior immunogenicity and long-lasting protection at lower antigen doses. It elicits balanced Th1/Th2 cytokine responses and demonstrates durable efficacy in both immunocompetent and complement-deficient mouse models. The platform’s compatibility with clinically approved adjuvants (MPLA and QS-21), modular peptide design, and potential for multi-pathogen applications underscores its translational promise. FM-SNAP exemplifies a next-generation vaccine strategy that is both scalable and adaptable for high-risk immunocompromised populations. Full article