Search Results (1,220)

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

Oak trees are widely distributed nationwide and account for approximately 24% of the total forest area in South Korea. However, these species are currently threatened by oak wilt disease caused by Dryadomyces quercus-mongolicae, leading to significant economic and ecological losses in the forestry industry. This study evaluated the effectiveness of culture suspension and lyophilized powder formulations of Streptomyces blastmyceticus in controlling oak wilt disease on Mongolian oak (Quercus mongolica). Field experiments were conducted using trunk and root injection methods in Q. mongolica plantations. The non-conductive area (NCA) of sapwood and colonization rate of the oak wilt fungus were analyzed and compared across treatments. In the Chuncheon experiment, Kangwon province, only the root injection of fungicide showed a significant difference compared to the culture suspension treatments. There were no significant differences between culture suspension and lyophilized powder treatments in Uiwang, Gyeonggi Province. Specifically, both preventive and curative treatments using culture suspension and lyophilized powder of S. blastmyceticus resulted in significantly different NCA values compared to the negative control (8.7%) and positive control (88.5%). The NCA for culture suspension ranged from 33.3% to 49.9%, and for lyophilized powder, from 37.3% to 43.9%. The colonization rate of the oak wilt fungus was lowest (9.72%) in the preventive treatment using lyophilized powder via trunk injection. For the culture suspension, the lowest colonization rate (20.83%) was observed in the curative treatment using trunk injection. These findings suggest that the lyophilized powder formulation of S. blastmyceticus efficiently suppresses the progression of oak wilt disease under field conditions. Full article

Background: Thermostability remains a key bottleneck for equitable access to mRNA-LNPs vaccines, mainly due to cold-chain requirements. Objectives and methods: Here, we optimized freeze-drying formulations by screening excipients (sugars, sugar alcohols, and proteins) and buffers to preserve mRNA-LNPs as solid formulations under ambient and refrigerated conditions. Physicochemical properties (size, polydispersity index [PDI], and encapsulation efficiency [EE]) and functional integrity, assessed by fluorescence-based in vitro transfection assays, were evaluated during long-term storage of up to six months. Results: Preliminary screening identified 20% sucrose and trehalose with Tris or histidine buffers as optimal for preserving physicochemical properties during freeze-drying, including high encapsulation efficiency (>90%), particle size (~200 nm), and low polydispersity (PDI < 0.2). Mannitol, gelatin, and PBS-based buffers showed adverse effects. At 4 °C, formulations F1–F3 maintained physicochemical stability and functional transfection activity for up to four months. In contrast, 20 °C storage caused progressive destabilization, with increased size, PDI, and encapsulation loss (>60% by six months). Among all formulations, 20% sucrose with 5 mM Tris (F1) showed the most robust preservation of physicochemical integrity and in vitro transfection efficiency under refrigerated and ambient conditions. Conclusions: Sugars outperformed sugar alcohols and gelatin as cryoprotectants. All formulations were stable, including functionally active at 4 °C for up to four months, while a sucrose/Tris formulation retained acceptable stability at 20 °C. Overall, the results demonstrate the feasibility of storing mRNA drug products as solid formulations at non-freezing temperatures. Full article

Background: In this study, we aimed to compare metabolomic profiles, biodistribution, and detoxification patterns of the novel SN-38 derivative NMe with irinotecan (IR), and to identify NMe-specific metabolites to evaluate its preclinical pharmacokinetic advantages. Methods: In vivo ADME studies were conducted for NMe, a 9-aminomethyl SN-38 derivative, and IR following a single intraperitoneal dose of 40 mg/kg in mice. Additionally, ADMET properties were predicted using ADMETlab and SwissADME tools for comparison. Levels of NMe and irinotecan absorbed into plasma, distributed to tissues, and metabolized were monitored in liver, lung, spleen, kidney, and stool samples at 15, 30, and 60 min post-administration. Tissue extracts were analysed using high-performance liquid chromatography (HPLC), liquid chromatography–electrospray ionization quadrupole time-of-flight-tandem mass spectrometry (LC-ESI-QTOF-MS), and nuclear magnetic resonance (NMR) techniques after lyophilization and reconstitution. We compared the metabolomic profiles of irinotecan and NMe. Results: We identified and confirmed NMe-specific metabolites, including 9-CH₂-S-cysteine conjugate, 9-CH₂OH, and NMe-formyl. Notably, novel irinotecan metabolites (IR-OH and IR-ΔE) were detected in small amounts in kidney samples. In some cases, two literature-known photodegradation products of irinotecan were present. NMe was found to quickly metabolize with different distribution to tissues, significantly greater to kidney and liver. Two SN-38 glucuronides, SN-38G(α) and SN-38G(β), were detected corresponding to α- and β-anomers. Where it was possible, NMe, IR and SN-38 were quantified using external calibration curves. In IR group, controlled and prolonged release of SN-38 was confirmed in all samples, yet SN-38G was observed in minority only in plasma, kidney, or lungs. In NMe groups, great relative amounts of SN-38 and SN-38G were detected. Greater content of SN-38G in NMe group than in irinotecan is expected to contribute to modulation and alleviation of some side effects in irinotecan-involved therapies, such as gastrointestinal toxicities (GIT). Conclusions: NMe shows a distinct metabolic profile characterized by rapid biotransformation, higher systemic glucuronidation of SN-38, and formation of unique metabolites, suggesting a potentially wider therapeutic window and reduced toxicity compared with IR. Full article

Backgrounds: Antibody–drug conjugates (ADCs) combine the specificity of monoclonal antibodies with the cytotoxic potency of drugs, representing a significant class of targeted cancer therapeutics. Despite their clinical success, formulation-related instability, rather than biological inefficacy, is a major contributing factor to setbacks in ADC development. This review examines the biochemical, physicochemical, and formulation factors that contribute to ADC stability, with a focus on excipient selection, conjugation site heterogeneity, and linker–payload reactivity. Methods: This comprehensive review was based on a selection of peer-reviewed mechanistic, analytical, and manufacturability studies on ADC stability. Our goal was to highlight formulation strategies, degradation pathways, and solid-state stabilization principles that affect the pharmacokinetics and therapeutic efficacy of ADC. Results: Results demonstrate how formulation variability including buffer composition, excipient choice, ionic strength, and lyophilization can directly affect payload release, linker cleavage, kinetics, and antibody conformation. It has been demonstrated that techniques, such as lyophilization with glass-forming matrices and the addition of surfactants, enhance stability against hydrolysis, oxidation, and aggregation. Developments in analytical characterization, such as real-time kinetic modeling and multi-attribute techniques based on mass spectrometry, have made quantification of degradation and bioactivity losses more predictable in ADC formulations. The connection between chemical stability and formulation outcomes is being redefined by new techniques, such as model-informed optimization and AI-driven design. Conclusions: ADC formulation is now a key component of molecular stability, clinical reliability, and regulatory compliance rather than a secondary consideration. By guaranteeing long-term stability, better pharmacokinetics, and improved therapeutic indices across next-generation designs, these approaches have the potential to revolutionize ADC development. Full article

With rising global temperatures, biostimulants might be a promising tool to alleviate plant stress and support adaptation. The potential of fresh (FBP) and lyophilized (LBP) banana peel aqueous extracts as biostimulants for protecting broccoli from high temperature (HT) stress was analyzed. Spectrophotometric and statistical analyses revealed that BP affected broccoli phytochemistry in a temperature-dependent manner. Under room temperature (RT), FBP and LBP decreased glucosinolates (−15% and −25%, respectively). Conversely, FBP increased flavonols and proanthocyanidins (141% and 202%, respectively). Under RT, LBP decreased sugars in broccoli (−27%). FBP had stronger effects at HT than at RT, further boosting phenolics (70%), flavonoids (89%), tannins (31%), and hydroxycinnamic acids (64%), and antioxidant capacity (FRAP) (10%). LBP also increased flavonoids (39%), flavonols (95%), and hydroxycinnamic acids (45%) under HT. Both FBP and LBP increased glucosinolates (47% and 46%, respectively) in HT-grown broccoli. HT significantly affected glucosinolates, decreased them in control plants, and increased them in BP-treated plants. All HT-grown plants had higher soluble sugars and lower hydrogen peroxide than RT-grown plants. Principal component analysis confirmed greater biochemical diversity under HT. Temperature–BP interaction significantly affected flavonoids and glucosinolates, highlighting the central role of environmental temperature in determining biostimulant outcomes. These findings suggest that global warming may markedly alter biostimulant efficacy and should be considered in their development. Full article

Background/Objectives: Immune-mediated hepatitis, including autoimmune hepatitis, remains a formidable clinical challenge characterized by the rapid destruction of the liver parenchyma. While whey proteins are well-regarded for their anti-inflammatory properties, goat whey possesses a distinct bioactive profile, offering superior digestibility and reduced allergenicity compared to their bovine counterparts. This study investigated the hepatoprotective potential and underlying immunological mechanisms of lyophilized goat whey (LGW) in a Concanavalin A (ConA)-induced model of acute hepatitis. Methods: BALB/c and C57BL/6 mice were administered LGW orally (1 g/kg/day) for five consecutive days prior to a ConA challenge. Liver injury was quantified via serum transaminase levels and histopathological evaluation. The cytokine profiles and the phenotype of liver mononuclear cells (MNCs) were analyzed using ELISA and flow cytometry, respectively. Results: LGW pretreatment significantly attenuated ConA-induced hepatitis in both mouse strains, markedly reducing serum transaminase levels and preserving hepatic architecture. Mechanistically, LGW triggered a fundamental shift in the hepatic immune microenvironment by suppressing the pro-inflammatory Th1/Th17 axis (evidenced by decreased IFN-γ and IL-17) while concurrently upregulating the anti-inflammatory cytokine IL-10. Furthermore, LGW induced a tolerogenic phenotype in hepatic dendritic cells (CD11c⁺CD206⁺), which directly correlated with a significant expansion of regulatory T cells (Tregs). This strain-independent protection suggests that LGW modulates fundamental, early-stage immune signaling pathways within the liver. Conclusions: Our findings demonstrate that LGW exerts potent hepatoprotection by effectively reprogramming the hepatic immune microenvironment toward a tolerogenic state. These results position LGW as a promising, safe, and effective functional food candidate for the prevention and adjunct management of immune-mediated inflammatory liver diseases. Full article

The medical field now uses mRNA therapeutics to deliver fast programmable treatment options through versatile vaccination platforms. The worldwide adoption of mRNA therapeutics faces a major obstacle because these molecules require extreme cold storage and transportation systems. mRNA stability establishes a fundamental scientific and industrial challenge which requires researchers to unite formulation design with process control and material engineering for cold-chain independence. Current knowledge about RNA hydrolysis and lipid oxidation and water-mediated degradation is combined with new methods for solid-state stabilization through lyophilization and spray-freeze-drying and thin-film technologies. Mechanism such as vitrification, water replacement and excipient RNA interactions are assessed to establish the fundamental chemical properties needed for extended product stability. Advanced mRNA development strategies are also examined, including self-amplifying and circular RNA structures and nano-glass and metal–organic frameworks and artificial intelligence-based predictive design for creating stable mRNA formulations at room temperature. This review examines manufacturing and regulatory and logistical obstacles which affect real-world implementation of mRNA therapeutics through assessments of production scale and product quality tests and packaging strength and tropical environment testing. The combination of research findings presents a path to develop mRNA medicines which maintains their effectiveness when stored at 25 °C or above, thus enabling worldwide access to RNA-based treatments. The development of mRNA into a durable therapeutic platform requires scientists to merge molecular research with process development and regulatory standardization. Full article

Mesenchymal stem cells (MSCs) are candidates for the treatment of palatal wounds in combination with biomaterials. In this study, we developed a method for the production of a ready-to-use mucoadhesive hydrogel containing MSCs for palatal wounds and evaluated its healing effects. Dental follicle MSCs (DFMSCs) were isolated from the dental follicle tissue of a healthy twenty-year-old donor. DFMSCs were suspended in a cell-preserving solution containing platelet-rich plasma, trehalose, and DMSO, and loaded into a catechol–chitosan hydrogel solution at a ratio of 1:400 (v/v) with 5 × 10⁵ or 6 × 10⁶ cells per hydrogel to create a novel lyophilization method for cell integration into the biomaterial. Hydrogels were fabricated as scaffolds with a diameter of 5 mm and a depth of 4 mm. After lyophilization of the hydrogels with cells, a viability test was performed after the production of hydrogels on the seventh day and the fifth month. Palatal wounds were created as full-thickness wounds in rats using a 5 mm diameter punch. The hydrogels were applied to the palatal wounds of rats, and histochemical and immunohistochemical analyses were performed. The results showed that, after rehydration of the hydrogels, DFMSCs had over 80% viability and were homogeneously distributed in the hydrogels. After the application of DFMSC-loaded hydrogels, palatal wounds healed within 7 days, and inflammatory cell infiltration, fibroblastic proliferation, and ulceration were significantly reduced, while epithelial regeneration was significantly increased compared to the control group. The viability ratio of DFMSCs was 83.7% on the seventh day and 71.3% in the fifth month. Hydrogels loaded with DFMSCs represent a promising, ready-to-use biomaterial approach for supporting palatal wound healing. Full article

Freeze-drying, also known as lyophilization, is a state-of-the-art method for preserving food, offering excellent retention properties for nutrients, structure, and taste compared to other drying processes. Freeze-drying yields a product visually similar to fresh produce. However, due to the high energy requirements and operational costs associated with the process, its broader use as an industrial tool is limited. This review encompasses the optimization of all key stages, including pretreatment, freezing, primary drying, secondary drying, and storage. Process efficiency and product quality depend on a variety of factors, including raw material composition, pretreatment strategies (e.g., Pulsed Electric Fields), chamber pressure, shelf temperature, and freezing rate. These parameters are critical control points for determining the final product outcome. Optimizing these parameters is essential; as summarized by recent research, lyophilization effectively protects bioactive compounds, color, flavor, and rehydration ability in various food systems, including fruits, vegetables, meats, seafood, and specialty products. To achieve broader industrial adoption, this gold-standard method requires advancements in process intensification and hybrid drying systems, potentially integrated with intelligent process control. These advances are crucial to enhancing the economic viability of freeze-dried products and maintaining their reputation as the gold standard in creating high-quality, shelf-stable food products. This review consolidates current knowledge into a coherent conceptual model. The model clarifies the deterministic sequence by which adjustable processing conditions direct essential physicochemical changes within the food matrix, thereby defining the product ultimate nutritional, sensory, and stability properties. Establishing this cause-and-effect framework provides a foundation for systematic process improvement and facilitates broader commercial implementation. Full article

Although blood–brain barrier (BBB) models are of great value in investigating neurological diseases, the structural complexity and intricate function based on cell–cell interactions of the BBB bring various limitations to the applications of existing models. In this study, a novel BBB micro-organoid model was established by culturing neurovascular unit (NVU) cells on a decellularized squid mantle scaffold (DSMS) film to reconstitute a more authentic and reliable NVU microenvironment for in vitro research. The DSMS applied was obtained from squid mantle scaffolds via decellularization, followed by defatting, and showed good biocompatibility with no cytotoxicity. The DSMS film was finally prepared by lyophilization. The lyophilized film exhibited a void ratio and pore size suitable for the adhesion and growth of endothelial cells (hCMEC/D3) and astrocytes (hACs), which led to the formation of a BBB-like spatial structure. The BBB micro-organoid model exhibited functional barrier properties, including an effective transendothelial electrical resistance (TEER) of approximately 230 Ω/cm², restricted permeability to macromolecules—with apparent permeability coefficients (Papp) of 6.3 × 10⁻⁷ cm/s for 10 kDa and 2.7 × 10⁻⁷ cm/s for 70 kDa FITC–dextran—and expression of tight junctional complex (TJC) proteins such as vascular endothelial cadherin (VE-cad) and Zonula Occludens-1 (ZO-1). Furthermore, low-density lipoprotein receptor-related protein 1 (LRP1), a key receptor stably expressed in these two NVU cell types, was utilized as a critical indicator to assess the integrity of the BBB micro-organ model and its responsiveness to pathophysiological stimuli, particularly under thrombotic conditions. This study not only validates the feasibility of constructing a functionally competent BBB micro-organ model using DSMS films integrated with NVU cells but also provides a promising in vitro platform for subsequent studies on the BBB-related pathological mechanisms and the evaluation of drug permeability across the BBB. Full article

The milk metabolite profiles of dairy cows during the dry-off and peripartum periods were investigated using ¹H NMR combined with chemometric analysis to evaluate the effects of different dry-off management strategies. Milk samples were collected 14 days before dry-off (T0) and 28 days after calving (T1) from cows receiving an internal teat sealant combined with intramammary antibiotics (CTR), an internal teat sealant alone (SIG), or an internal teat sealant associated with dietary supplementation of lyophilized Aloe arborescens (ASIG). Analysis of both aqueous and organic milk extracts revealed no significant metabolite differences among treatment groups. In contrast, a clear discrimination was detected between samples collected at T0 and T1. Aqueous extracts at T0 were characterized by higher levels of choline, butyrate, branched-chain amino acids, and N-acetylated compounds, whereas T1 samples exhibited higher levels of saccharides, citrate, phosphorylcholine, and galactose-1-phosphate. Organic extracts at T0 showed higher concentrations of conjugated linoleic acids (CLAs) and caproleic acid. These findings indicated that the physiological stage of the cows had a more pronounced impact on milk metabolite composition than the dry-off treatments, with no detrimental effects on milk composition or overall metabolite balance. Full article

In recent years, insect-derived peptides have attracted attention for their potential biological activities, particularly antioxidant properties. This study assessed the antioxidant activity of two widely consumed edible insects, T. molitor and A. diaperinus larvae, using cell-free and cell-based approaches. Whole lyophilized larvae, digestion products from the oral, gastric, and intestinal phases, as well as the <3 kDa permeate fraction (D-P3) derived from the intestinal digestion phase, were evaluated using biochemical antioxidant assays. Overall, digested samples exhibited higher antioxidant capacity than their undigested counterparts. At the cellular level, treatment of LPS-stimulated, PMA-differentiated THP-1 macrophages with A. diaperinus D-P3 was associated with increased mRNA expression of genes related to antioxidant defense, including NFE2-like bZIP transcription factor 2 (NFE2L2, also known as Nrf2), glutathione-disulfide reductase (GSR), superoxide dismutase 1 (SOD1), and catalase (CAT), whereas T. molitor D-P3 preferentially modulated nuclear factor kappa B p50 subunit (NFKB1) and nuclear factor kappa B p65 subunit (RELA). Overall, these findings indicate that gastrointestinal digestion enhances the bioaccessibility of antioxidant components in both edible insect species while revealing species-specific transcriptional responses under in vitro inflammatory conditions. This multilevel assessment provides mechanistic insight into the antioxidant-related biological activity of digestion-derived insect peptides and supports their further investigation as functional ingredients in food and feed systems. Full article

Nucleation remains one of the least understood steps during protein crystallization, although it strongly impacts product quality attributes, including total crystal numbers, final crystal size distributions, and thus downstream processing. In this work, the nucleation behavior of Lactobacillus brevis alcohol dehydrogenase (LbADH) wild type (WT) and five mutants (Q207D, Q126H, K32A, D54F, and T102E) is investigated in a stirred 7 mL crystallizer monitored by in situ multi-angle dynamic light scattering (MADLS). Nucleation was studied with highly pure homotetrameric LbADHs by establishing a crystallization, lyophilization, and re-solubilization protocol combined with size exclusion chromatography (SEC) and size exclusion high-performance liquid chromatography (SE-HPLC), yielding tetramer purities above 94% and removing low molecular weight impurities. During stirred batch crystallizations initiated by the addition of polyethyleneglycol 550 monomethyl ether (PEG 550 MME), SEC and SE-HPLC revealed decreasing tetramer peak areas but essentially constant peak apex positions, indicating that no long-lasting oligomeric intermediates accumulate at detectable levels. Time-resolved MADLS measurements using a custom-made flow-through cuvette in a bypass to the stirred crystallizer uncovered transient cluster populations. All protein variants exhibited an initial tetramer peak, followed by the formation of larger aggregates and a rapid rise in signal above a hydrodynamic diameter of 1000 nm, coinciding with the onset of macroscopic turbidity. A simple mesoscale nucleation model was formulated, yielding end-of-nucleation times, crystallized fractions, critical soluble concentrations, and apparent nucleation rate constants. The crystal contact mutations modulate both the timing and magnitude of the nucleation burst (rapid build-up of nuclei/cluster populations). The mutant Q207D showed strongly attenuated nucleation compared to the WT, whereas the other mutants (K32A, D54F, and particularly T102E) display markedly accelerated nucleation at nearly invariant critical concentrations. The combined workflow demonstrates how in situ MADLS, together with a tailored kinetic description, can provide mechanistic insight into protein nucleation in stirred batch crystallizers. Full article

Background/Objectives: Rapid diagnostic tests (RDTs) for human immunodeficiency virus (HIV) are widely used, but most kits lack standardized internal quality control (IQC) materials. In this study, we aimed to develop and evaluate a plasma-based IQC compatible with five HIV RDT brands and with proven long-term stability. Methods: Control samples at three reactivity levels were tested with five HIV RDT kits in lyophilized and liquid forms. Lyophilized samples were produced with and without trehalose, whereas liquid samples were prepared with and without StabilZyme™ SELECT Stabilizer (Stabilizer). Accelerated stability testing was performed at 37 °C and 45 °C for 28 days, and the most stable formulation was selected for long-term storage at 4 ± 2 °C and 25 ± 5 °C. Stability was assessed based on test-line visibility and signal intensity. Signal-intensity trends were analyzed using simple linear regression with a t-test on the slope; samples were considered stable when no significant trend was detected (p > 0.05). Results: Reactivity measured using the Elecsys HIV combi PT assay yielded cutoff index (COI) values of 772.65 (1:8) for the strong-positive control and 269.95 (1:25) for the weak-positive control. Trehalose-containing lyophilized samples maintained reactivity under accelerated testing at 37 and 45 °C and for 6 months at 4 ± 2 °C and 25 ± 5 °C, with no significant change in signal intensity (p > 0.05). Conclusions: The plasma-based IQC materials were compatible with all five HIV RDTs, and trehalose-stabilized lyophilized plasma showed high stability, supporting transport and storage without strict cold-chain requirements. Full article