Search Results (1,234)

Lipid nanoparticles (LNPs) have become the cornerstone of nucleic acid delivery platforms, particularly in RNA-based vaccines and therapeutics. However, the conventional methods of LNP production, which are primarily reliant on microfluidic mixing of aqueous and organic solvent phases, pose limitations in terms of mRNA stability, residual organic contamination, scalability, cost, and environmental impact. These limitations prompted a renewed search for non-conventional strategies with the promise of improving mRNA-LNP encapsulation approaches. These emerging approaches aim to address key bottlenecks, including mRNA hydrolysis-driven degradation, high production losses, and complex downstream purification. Moreover, the ability to decouple LNP synthesis from mRNA encapsulation could enable streamlined, modular manufacturing workflows and customizable payload delivery, including single- or multiple-mRNA payloads, thereby expanding the therapeutic scope of LNPs. This review offers an early insight into the design principles and scalability potential of emerging non-conventional LNP encapsulation approaches, including solvent-free and microfluidics-free methodologies, and pre-built LNP workflows. We also examine trends in emerging LNP encapsulation tools, including high-shear mixing, sonication, membrane contraction, and other approaches. Finally, we extrapolate the suitability of the methods for scale-up approaches and their economic implications based on the process information. Full article

This narrative review examines the clinical application of tiletamine–zolazepam (TZ) in exotic pet and wildlife anesthesia, addressing the complexities inherent in managing a broad taxonomic spectrum with diverse physiological profiles and temperaments. As a fixed-dose combination, TZ is a cornerstone of multimodal protocols designed to achieve balanced anesthesia. Its lyophilized formulation permits reconstitution with various sedative solutions, facilitating low-volume administration, a critical requirement for the immobilization of wildlife and small exotic patients. Given the significant variability in species-specific responses and environmental influences, selecting and adapting appropriate TZ-based protocols remain a challenge for practitioners. By synthesizing heterogeneous data into expert-validated guidance, this review provides specialized and general veterinarians with practical considerations for the judicious use of TZ. Emphasis is placed on integrating TZ within multimodal protocols to mitigate arousal risks, ensure consistent immobilization, and facilitate rapid recovery. This approach seeks to optimize anesthetic outcomes and promote animal welfare across these physiologically diverse populations. Full article

Background/Objectives: Lipid nanoparticles (LNPs) containing ionizable lipids represent the most advanced non-viral delivery vehicles and have become state-of-the-art carriers for RNA therapeutics. However, further improvements in endosomal escape efficiency and biodegradability are still needed, especially for nucleic acids with transient activity such as messenger RNA (mRNA) and transfer RNA (tRNA). Methods: In this study, a novel library of highly biodegradable ionizable lipids featuring thioester groups within the linker region was designed and synthesized, thereby expanding the chemical linker toolbox for future ionizable lipid development. Results: Comprehensive in vivo structure–activity relationship studies led to the identification of CP-LC-1272 as a lead candidate that markedly enhances endosomal escape and exhibits superior in vivo biodegradability, attributed to the high acid-lability of thioester bonds. LNPs containing CP-LC-1272 maintained in vivo activity after six months of storage in lyophilized form and demonstrated superior in vivo efficiency compared to SM-102 in mRNA expression studies, as well as similar protein restoration in a tRNA delivery model targeting premature stop-codon mutations. Conclusions: The rapid biodegradability of these thioester-activated ionizable lipids (TAILs) suggests a reduced risk of accumulation, with the potential to enable safe repeated dosing or high-dosage RNA therapies, positioning TAILs as a versatile and safe platform for next-generation RNA therapeutics. Full article

In this study, Paliperidone Palmitate (PP), a second-generation antipsychotic, commonly used for the treatment of schizophrenia, was encapsulated in bio-based non-isocyanate polyurethane (NIPU) nanoemulsions. NIPU was synthesized via an isocyanate-free polyaddition route, addressing safety and environmental concerns associated with conventional polyurethanes. The drug-loaded nanoparticles were produced utilizing oil-in-water (O/W) emulsions followed by solvent evaporation and lyophilization. NIPU concentrations of 0.3% and 0.5% w/v, as well as 0.5% w/v PVA were employed, while PP was incorporated at 0.2%, 0.5% and 1% w/v. The formulations were characterized by FTIR, DSC and XRD analyses, and the mechanical strength of neat sponges was evaluated. The nanoparticle formation and size were assessed by DLS and SEM analyses. The water contact angle, porosity measurements and aquatic and enzymatic hydrolysis were additionally performed. The resulting nanocarriers exhibited controlled particle size, increased drug-loading values, structural stability and biodegradability. Lastly, the in vitro dissolution studies revealed a system-specific burst release behavior, and a controlled and sustained overall drug-release profile for majority of the formulations, thereby indicating the potential of NIPU nanocarriers for drug delivery applications, particularly where sustained therapeutic effects are required. Full article

Kombucha is a fermented beverage produced using a symbiotic consortium of acetic acid bacteria and yeasts, often marketed for its health-promoting properties. However, probiotic bacteria in kombucha are typically present at inconsistent levels and may not remain viable during fermentation. In this study, three Lactobacillus strains (Lacticaseibacillus rhamnosus ATCC 53103 (L. rhamnosus), Lactiplantibacillus plantarum subsp. plantarum ATCC 14917 (L. plantarum) and Lentilactobacillus hilgardii (L. hilgardii) isolate) were encapsulated in whey protein using the lyophilization method and added individually at the start of kombucha fermentation. Lactic acid bacteria (LAB)–enriched kombucha samples were evaluated for chemical composition (polyphenols, flavonoids, vitamin C and organic acids) and functional properties (antimicrobial, antiproliferative, antioxidant and anti-inflammatory activities) and compared to a traditionally obtained control kombucha, primarily demonstrating in vitro and experimental assessment. Encapsulation maintained LAB viability above 6–7 log CFU/mL throughout fermentation, producing kombucha with enhanced microbial stability. LAB–enriched samples exhibited increased L-lactic acid and antimicrobial activity. L. rhamnosus and L. hilgardii–enriched samples exhibited increased antiproliferative and anti-inflammatory activities, which may be associated with strain-dependent production of organic acids, polyphenol modulation and LAB-derived bioactive metabolites. Antioxidant activity varied depending on assay, and L. rhamnosus–enriched kombucha showed higher anti-inflammatory activity. These findings demonstrate that whey protein encapsulation can preserve LAB during fermentation, enhance specific bioactive properties and provide a platform for developing functional kombucha beverages with potential applications in the food industry. Full article

Phyllanthus emblica (amla) exhibits anticancer activity, but its extracts often suffer from poor stability and bioavailability. This study developed amla extract-loaded niosomes to enhance delivery and evaluate their anticancer activity against MCF-7 and HCT116 cell lines, supported by in silico analyses. Methodology: Amla extract was prepared using a 50% aqueous–alcoholic solvent system and lyophilized. Niosomes were prepared by the thin-film hydration method and characterized for physicochemical properties. Anticancer activity was evaluated through in vitro cytotoxicity studies, supported by molecular docking and in silico pharmacokinetic analyses. Results: Optimized niosomes exhibited spherical morphology, good homogeneity (PDI < 0.30), anionic surface charge, high entrapment efficiency (70.5 ± 5.9%), and sustained diffusion-controlled release. In vitro cytotoxicity demonstrated a strong concentration-dependent anticancer activity of amla-loaded niosomes across a range of concentrations (31.25–1000 µg/mL) against both MCF-7 and HCT116 cell lines. At 1000 µg/mL, cell viability decreased to 7.0% and 5.4% in MCF-7 and HCT116 cells, respectively, with calculated IC₅₀ values of 245 µg/mL and 158 µg/mL. Molecular docking and pharmacokinetic predictions supported the potential multi-target anticancer relevance of major phytochemicals, including hydrolyzable tannins, phenolic acids, flavonoid aglycones and glycosides, and highlighted bioavailability limitations for certain high-affinity glycosylated flavonoids, reinforcing the rationale for vesicular encapsulation. Conclusions: Amla extract-loaded niosomes represent a promising vesicular system for enhanced, sustained delivery of anticancer activity in vitro, with complementary in silico findings supporting mechanistic plausibility and translational rationale. Further studies are warranted to evaluate their performance in vivo. Full article

Gluten-free (GF) products developed for individuals with celiac disease and gluten sensitivity often suffer from low protein and mineral content. Fish proteins offer a promising solution to address these deficiencies; however, the characteristic “fishy odor” and related technological challenges limit consumer acceptance. This study aimed to develop an innovative GF pasta with improved nutritional density and acceptable sensory quality by incorporating deodorized and lyophilized trout powder. GF pasta formulations were prepared using buckwheat flour, xanthan gum, and 5% or 10% odorless trout powder. Vinegar pretreatment was applied to reduce fish odor, while lyophilization was chosen to minimize nutrient losses. The samples were analyzed for nutritional composition, techno-functional properties, in vitro digestibility, and sensory attributes. Results showed that trout powder significantly increased protein and ash content compared to the control (p < 0.05). A slight darkening was observed in color analysis due to fish pigments and buckwheat phenolics, but overall visual stability remained high. In vitro digestibility revealed enhanced protein digestibility (p < 0.05) and a slight reduction in starch digestibility. Sensory evaluation demonstrated that odor scores (8) at 10% trout inclusion remained close to the control, reversing the commonly reported decline in acceptance with increasing fish content. These findings indicate that combining vinegar pretreatment with lyophilization enables the incorporation of fish proteins into GF pasta without sensory disadvantages, while simultaneously improving nutritional quality. Full article

Effective oral interventions for alcohol-induced metabolic stress and liver injury remain limited. Pre-absorptive gastrointestinal alcohol handling is gaining interest as a non-pharmacological strategy to reduce hepatic burden. In this study, we developed a formulation-integrated, food-compatible lyophilized recombinant whole-cell catalyst based on Escherichia coli Nissle 1917 engineered to express alcohol dehydrogenase and acetaldehyde dehydrogenase. Rather than focusing exclusively on strain-level genetic modification, the engineered cells were protected by lyophilization combined with a food-grade chitosan–alginate layer-by-layer coating, forming an artificial cell wall designed to enhance survivability during oral delivery. The formulation resisted simulated gastric acid, sodium taurocholate, and ethanol, retained enzymatic activity after storage, and demonstrated formulation stability. In alcohol-exposed mice, oral administration reduced blood ethanol and acetaldehyde levels, improved liver biochemical parameters, attenuated hepatic steatosis, and partially restored oxidative stress indicators. Integrated multi-omics analyses indicated coordinated gut-associated metabolic and inflammatory responses to alcohol and intervention, rather than a single dominant pathway. These findings provide hypothesis-generating evidence; causality remains to be established. Overall, this study demonstrates a proof-of-concept, food-compatible lyophilized recombinant whole-cell catalyst that integrates enzymatic function with formulation stability and gastrointestinal resilience, highlighting an applied, food-compatible microbial framework for exploring alcohol-related metabolic stress. Full article

Skin pigmentation is a crucial factor influencing the market value of gilthead seabream. A three-month feeding trial evaluated the effects of dietary inclusion of Rhodotorula mucilaginosa on skin and fillet pigmentation of gilthead seabream (Sparus aurata). Four diets containing yeast were tested in triplicate tanks using 120 fish in total. Skin and fillet colours were assessed via computer-based image analysis in CIELAB, RGB and HSB spaces. Analysis of total carotenoids was also performed. Yeast inclusion increased L* and Whiteness values in the operculum and enhanced lightness in ventral skin regions. In the abdominal area, RGB values decreased, particularly in the 3% diet. Fillet responses were limited to the red muscle, where the 3% diet significantly increased a* and Chroma values. Overall, inclusion of R. mucilaginosa exceeding 2% influenced seabream skin brightness and total carotenoid content, while 3% inclusion enhanced red muscle pigmentation, suggesting potential as a natural pigment source in seabream feeds. Full article

Paclitaxel is a first-line anticancer drug, but its low water solubility impedes bioavailability. The purpose of this study is to estalish a delivery strategy via carboxymethyl chitosan (CMCS)-encapsulated 6-deoxy-6-mercapto-β-cyclodextrins (dmβCDs)-modified concave cubic gold (CCGs) to achieve PTX release. CCGs were initially synthesized by the one-pot method and further modified by dmβCDs, the dmβCDs can effectively capture PTX molecules, followed by encapsulation with CMCS, and then prepare pH-responsive CMCS/dmβCDs/CCGs nanocarriers after lyophilization. Results indicated that desirable hexagonal CCGs with 50 ± 5 nm size can be obtained by adjusting H₂O₂ and HClO concentration. FT-IR, Raman and XRD spectra had confirmed dmβCDs successfully grafted to the surface of CCGs. Drug loading experiments demonstrated that the nanocarrier encapsulated PTX in amorphous powder or molecular form have a capacity of 55.05 µg/mL. Drug release experiments revealed PTX release from CMCS/dmβCDs/CCGs nanocarriers carrying a typical pH-responsive profile and indicating earlier release in an acidic environment than in a neutral or alkaline environment. The proposed method can be utilized to effectually achieve high-efficiency solubilization and targeted release inside tumor cells of PTX. Full article

Background/Objectives: Sea urchin gonads (“roe”) are a valuable seafood product and a chemically complex matrix whose composition varies with physiology and environment. We present a biphasic extraction and ¹H NMR workflow to build a reusable reference inventory of polar metabolites and apolar lipid features in Paracentrotus lividus. Methods: Gonads from 37 adults (23 males, 14 females) collected at two sites (Alicante and Jávea–Dénia, Spain; October 2024) were lyophilized, extracted with methanol/chloroform/water, and analyzed by 400 MHz ¹H NMR in buffered aqueous solution (polar) and CDCl₃ (apolar). Polar metabolite identification combined 1D patterns with database matching and ¹H–¹³C HSQC confirmation on representative samples, yielding 71 annotated resonances corresponding to 37 metabolites spanning amino acids, osmolytes/quaternary amines, carbohydrates/aminosugars, and nucleoside/purine-related compounds. Results: Polar fingerprints enabled supervised modelling: PLS-LDA separated sexes with low cross-validated error, and SPA/COSS ranking highlighted glycine, alanine, creatine and osmolyte-associated signals as key discriminants; pathway mapping supported the enrichment of amino-acid and one-carbon/purine networks. Apolar spectra were annotated at the motif level and used for lipid-index estimation, indicating substantial unsaturation but low docosahexaenoic acid (DHA) and modest sex effects. Conclusions: The curated peak lists and reporting framework facilitate reproducible NMR annotation and future comparative studies of P. lividus gonads. Full article

Probiotics hold considerable promise for treating and preventing inflammatory disease; however, their application is often limited by unclear anti-inflammatory mechanisms and reduced viability following lyophilization. In this study, I thoroughly evaluated the anti-inflammatory potential of Lactiplantibacillus plantarum FS4722 (L. plantarum FS4722) and substantially enhanced strain viability through optimization of the lyoprotectant formulation. Functional assays demonstrated that the fermented supernatant, heat-inactivated bacterial suspension, and cell lysate derived from L. plantarum FS4722 effectively suppressed transcription and expression of inflammatory cytokines in LPS-stimulated RAW 264.7 macrophages. The fermented supernatant exhibited the strongest inhibitory effects, surpassing the reference probiotic Lacticaseibacillus rhamnosus GG (LGG). Mechanistic investigations revealed that anti-inflammatory activity is primarily mediated via inhibition of the MAPK and NF-κB signaling pathways. Furthermore, using component screening combined with response surface methodology, the lyoprotectant formulation (10.00% trehalose, 1.00% sodium carboxymethyl cellulose, and 5.00% skim milk) was optimized, resulting in a lyophilization survival rate of 82.32% while maintaining cellular integrity; in this accelerated stability assessment, the strain retained 78.89% of its activity after 28 days of storage at 4 °C. Collectively, this study provides a robust and efficient approach for probiotic formulation while systematically elucidating the underlying anti-inflammatory mechanisms, thereby offering practical guidance for the development and clinical application of high-performance probiotic products. Full article

This work presents a method to encapsulate plant extracts within a binary polysaccharide carrier and to characterize the physicochemical and rheological performance of the resulting biocomposites in the context of food use. Using a starch/psyllium matrix, extracts from Sambucus nigra (SN), Aronia melanocarpa (AM), and Echinacea purpurea (EP) were effectively protected and incorporated through a stepwise workflow encompassing matrix preparation, encapsulation, structural verification, and functional assessment. SEM revealed a porous network containing uniformly distributed, extract-loaded spherical structures (~800–1500 nm), while FTIR supported the presence of hydrogen bonding and hydrophobic interactions that contributed to system stability. The prepared nanoemulsions showed shear-thinning (pseudoplastic) behavior, indicating favorable processing characteristics, whereas most physicochemical and bioactivity measurements were performed on lyophilized composites. The dried materials preserved extract-specific color signatures (ΔE > 5) and exhibited distinct thermal responses: AM produced a pronounced plasticizing effect (Tg reduced by >20 °C), while the incorporation of extracts generally delayed thermal degradation, consistent with polyphenol–starch interactions. Phase-transition behavior was also altered, with melting peaks suppressed for SN and AM and melting temperatures lowered for EP. Surface analysis indicated increased hydrophobicity and a reduced polar component of surface free energy, suggesting improved moisture barrier potential. Antioxidant capacity closely tracked total phenolic content (r > 0.94), with caffeic acid contributing strongly, particularly in EP-based systems. Antimicrobial activity depended on extract type (broad-spectrum for EP, selective for SN, minimal for AM), and the comparatively higher sensitivity of Gram-negative bacteria points to improved phenolic availability and membrane interactions upon encapsulation. Collectively, these results highlight the starch/psyllium matrix as a flexible platform for stabilizing plant extracts while enabling tunable functional attributes for functional food applications. Full article

Whole-grain rice (Oryza sativa L.) is a rich source of polyphenols. The in vivo mechanisms linking its phytochemical profile to antioxidant and anti-inflammatory effects remain incompletely defined. This study investigated the antioxidant and anti-inflammatory activity of a whole-grain rice flour 70% ethanol extract (OSEE) and correlated these effects with its phenolic composition. OSEE showed high total phenolic content 0.121 ± 0.002 mg GAE/g d.w.), a lower total flavonoid content (61.83 ± 4.03 µg QE/g d.w.), and a phenolic profile dominated by phenolic acids (~87%), with ferulic and protocatechuic acids among the most abundant constituents. OSEE displayed significant in vitro antioxidant activity in DPPH, FRAP, hydrogen peroxide, and nitric oxide scavenging assays. In vivo activity was evaluated in male Wistar rats with turpentine oil-induced acute inflammation using both therapeutic (post-induction) and prophylactic (pre-induction) protocols, testing three oral doses of lyophilized extract (1.0, 0.50, and 0.25 g/kg/day). In vivo, OSEE attenuated systemic oxidative stress (TOS, TAC, OSI, AOPP, MDA, NOx, 3-nitrotyrosine, total thiols) and the expression of pro-inflammatory markers (NF-κB p65, IL-1β, IL-18, caspase-1) in a dose-dependent manner with both protocols, with the highest dose producing the most consistent reductions, while the expression level of the anti-inflammatory factor IL-10 remained unchanged. PCA supported a shift in biomarker networks toward a non-inflamed profile. These findings indicate that OSEE exerts coordinated antioxidant and anti-inflammatory effects in vivo that are strongly associated with its phenolic composition. Full article

Collagen is a major extracellular-matrix protein widely used in regenerative medicine, yet conventional terrestrial sources raise biosafety and acceptability concerns, motivating the search for marine alternatives. This study evaluates the jellyfish Rhizostoma pulmo (R. pulmo) from the Azov Sea as a sustainable collagen source and assesses its suitability for biomedical materials. Acid-soluble collagen was extracted using 0.5 M acetic acid and purified by salt precipitation and dialysis, followed by physicochemical/structural characterization (sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE), Limulus amebocyte lysate (LAL) endotoxin testing, transmission electron microscopy (TEM), and immunofluorescence with type I collagen antibodies) and biological evaluation in vitro (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity on MRC5 fibroblasts; adhesion and proliferation assays on HeLa cells). The extracted collagen showed a high yield (~26.2%), a type I-like electrophoretic profile with α-, β-, and γ-components, fibrillar ultrastructure by TEM, and positive type I collagen immunoreactivity; endotoxin levels were low (0.461 EU/µL), and no cytotoxicity was detected under the tested conditions. Porous collagen sponges/scaffolds were fabricated by lyophilization, displaying interconnected pores with an average size of ~80 µm and pH-dependent swelling, and they supported 3D cell growth and tumor-cell dissemination in an in vitro breast carcinoma scaffold model. Overall, Azov Sea R. pulmo collagen demonstrates promising structural quality, low endotoxin burden, and cytocompatibility, supporting its potential as a marine biomaterial for sponge/scaffold-based tissue engineering and wound-related applications. Full article