Search Results (264)

The suckling phase is the critical window for rumen functional maturation, yet amino-acid-based interventions tailored to this stage remain scarce. L-citrulline (L-cit) bypasses hepatic first-pass metabolism, is converted to L-arginine peripherally, and resists ruminal microbial degradation, making it a candidate functional additive for early-life ruminants. This study evaluated whether dietary L-cit at 2 g·lamb⁻¹·d⁻¹ would improve rumen development and metabolic function in suckling Hu lambs. Twenty male Hu lambs were randomly assigned to a control (CON) or L-cit group (n = 10/group) and reared for 45 d (3 d adaptation + 42 d treatment). Growth and starter intake were assessed in all lambs; six lambs per group (n = 6) were subsequently slaughtered for rumen morphometry, gas chromatography–flame ionization detection (GC-FID) volatile fatty acid (VFA) quantification, 16S rRNA gene sequencing, and liquid chromatography–mass spectrometry (LC-MS) untargeted metabolomics. L-cit increased average daily starter intake by 25.96% (p = 0.036) and produced a 20.00% numerical but non-significant increase in average daily gain (ADG) (p = 0.203; Cohen’s d = 0.58). Rumen weight, volume, and papillary length, width, density, and epithelial thickness were all elevated (p < 0.05), whereas muscular thickness was unaffected (p = 0.162). Total VFA, acetate, propionate (+37.64%, p < 0.001), and butyrate were higher in the L-cit group; the molar proportion of propionate rose from 21.41% to 24.75%, and the acetate-to-propionate ratio declined from 2.90 to 2.44 (p = 0.005). Microbial richness (Chao1, Observed species) increased without altered evenness, and linear discriminant analysis effect size (LEfSe) identified L-cit-driven enrichment of propionate-generating and fiber-degrading genera, including Prevotellaceae_UCG-004, Ruminobacter, and the NK4A214_group. Of 539 differential metabolites (147 of which were annotated to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database), KEGG enrichment highlighted linoleic acid metabolism and purine metabolism as the biologically interpretable targets. Microbiota–metabolite correlations linked L-cit-enriched genera to up-regulated metabolites such as adenine. Dietary L-cit at 2 g·lamb⁻¹·d⁻¹ enhances starter intake, promotes rumen epithelial development, promotes a shift toward enhanced propiogenic fermentation within an acetate-dominant profile, and remodels the microbiota–metabolome axis, supporting its application as a functional additive during the suckling phase of ruminants. Because epithelial barrier integrity, oxidative stress, and inflammatory markers were not directly measured, these findings should be interpreted as morphological and association-based evidence, and further functional validation is required. Full article

Background: Oral delivery of chemotherapeutic agents remains challenging due to gastrointestinal degradation, poor intestinal permeability, and extensive first-pass metabolism, which collectively limit bioavailability. Lipid-based drug delivery systems offer a promising strategy to overcome these barriers. This study aimed to develop a freeze-dried, solid-dispersible self-emulsifying drug delivery system (SEDDS) using a water-in-oil-in-water (w/o/w) double emulsion approach for the co-encapsulation of hydrophilic (doxorubicin) and lipophilic (ellipticine) agents to enhance oral delivery. Methods: Double-emulsion SEDDS were prepared via a two-stage emulsification process to enable compartmentalized drug loading within aqueous and oil phases. The formulations were freeze-dried to improve stability and storage. Physicochemical properties were characterized using dynamic light scattering for droplet size and polydispersity index (PDI), zeta potential analysis for colloidal stability, and differential scanning calorimetry for thermal behavior. Drug encapsulation efficiency was determined, and cellular uptake was evaluated in breast cancer cells using fluorescence microscopy. Results: Optimized SEDDS exhibited droplet sizes of 90–347 nm with low PDI values (0.005–0.336), indicating uniform and stable dispersions. Zeta potential values (−10.64 to 2.38 mV) supported colloidal stability, while freeze-dried formulations retained dispersion characteristics upon reconstitution over extended storage. Both drugs demonstrated high encapsulation efficiency (>97%), and thermal analysis confirmed the formation of stable amorphous systems. Fluorescence imaging revealed enhanced intracellular uptake of both agents. Conclusions: This study demonstrates that freeze-dried double-emulsion SEDDS enable efficient co-delivery of hydrophilic and lipophilic drugs, improving stability and cellular uptake. This platform shows strong potential for overcoming key barriers in oral chemotherapy and provides a promising strategy for combination drug delivery. Full article

Icariin (ICA), a prenylated flavonoid glycoside from Epimedium (Yin Yang Huo), exhibits multi-organ pharmacological effects and has emerged as a promising candidate for osteoporosis therapy and bone tissue regeneration because of its capacity to modulate diverse osteogenic, anti-inflammatory, and angiogenic signaling pathways. Preclinical studies in osteoporotic models suggest that ICA improves trabecular microarchitecture and increases bone mineral density. Mechanistically, ICA modulates bone remodeling bidirectionally: it promotes osteoblast differentiation and extracellular matrix mineralization via activation of pro-osteogenic pathways, including Wnt/β-catenin and PI3K/Akt signaling, while simultaneously inhibiting osteoclastogenesis and bone resorption by suppressing RANKL-mediated NF-κB activation, thus reestablishing remodeling equilibrium. Despite these benefits, clinical advancement is hindered by the suboptimal oral bioavailability of ICA, stemming from poor intestinal absorption and extensive first-pass metabolism. To address this, innovative delivery systems have been engineered to enhance localized bioavailability and sustain therapeutic efficacy, such as hydrogel depots, nanoparticle formulations, and 3D-printed scaffolds enabling precise, controlled release. In bone tissue engineering applications, ICA-incorporated biomaterials—either standalone or in combination with osteogenic factors or exosomes—foster a regenerative niche by mitigating inflammation and oxidative stress, while synergistically promoting osteogenesis and angiogenesis, thereby expediting bone defect healing and osseointegration. Overall, these mechanistic elucidations and delivery advancements underscore ICA’s potential as a translational candidate for osteoporosis treatment and bone regenerative therapies. This review aims to critically and systematically synthesize current evidence on ICA-mediated bone repair and regeneration, with a particular emphasis on the molecular regulation of osteogenic signaling, the restoration of bone-remodeling homeostasis, and delivery-system-enabled strategies that may facilitate translational application. Full article

Maintaining hormonal equilibrium is a key determinant of women’s health, particularly during the menopausal transition and postmenopause. The decline in ovarian estrogen and progesterone production influences multiple physiological systems, affecting many aspects like vasomotor stability, bone and cardiovascular health, cognitive function, mood, and metabolic regulation. As a result, many women may experience symptoms that impair daily functioning and increase long-term morbidity. Recent progress in menopausal care emphasizes individualized, evidence-guided treatment, supported by improved diagnostic tools that allow for a more precise assessment of endocrine changes during this life stage. Among the available therapeutic options, transdermal menopausal hormone therapy has gained growing recognition due to its ability to re-establish hormonal levels with fewer systemic effects. By bypassing first-pass hepatic metabolism, this route provides more consistent serum hormone concentrations and may be associated with a lower risk of metabolic and thromboembolic complications compared with oral formulations. This review brings together the physiological basis, clinical indications, and current scientific evidence related to transdermal hormonal therapy during menopause while also highlighting its expanding therapeutic role and integration into personalized treatment strategies. In addition, we discuss recent findings on its pharmacological profile, clinical effectiveness, and emerging perspectives that position this therapeutic option as an increasingly important component of modern menopausal care and women’s health management. Full article

Background/Objective: Buccal delivery offers a potential route to circumvent gastrointestinal degradation and hepatic first-pass metabolism, but hydrophilic peptides typically exhibit limited mucosal permeation. Nanostructured lipid carriers (NLCs) have been proposed as delivery platforms capable of modulating interfacial interactions and improving mucosal transport. This study aimed to quantitatively evaluate the ex vivo buccal permeation of angiotensin II (Ang II), used as a hydrophilic peptide model, when associated with NLCs compared with free peptide under matched Franz diffusion cell conditions. Methods: Ang II-associated NLCs were prepared by melt emulsification combined with a low-energy injection technique. Particle size, polydispersity index, and zeta potential were determined by dynamic light scattering and laser Doppler electrophoresis. Association efficiency and drug loading were quantified by indirect spectrofluorometric analysis. Ex vivo permeation studies were conducted using porcine buccal mucosa mounted in Franz diffusion cells, and cumulative permeation, steady-state flux, and apparent permeability coefficients were calculated. Results: The NLCs exhibited nanometric size, moderate polydispersity, and association efficiency above 80%, and remained colloidally stable at 4 °C for 28 days. In ex vivo experiments, Ang II-associated NLCs showed measurable cumulative permeation, reaching approximately 9% after 2 h, whereas free Ang II was not detected in the receptor compartment under the tested conditions. Conclusions: This work provides a quantitative ex vivo buccal transport comparison of a hydrophilic peptide model delivered as NLC-associated versus free peptide under matched Franz cell conditions. The findings support further investigation of NLC-based approaches for buccal delivery of vasoactive peptides and provide a rational basis for future in vivo evaluation of mucosal delivery performance and systemic exposure. Full article

Hyaluronic acid (HA) is widely used in medical, cosmetic, and nutraceutical applications, yet the systemic fate of orally administered HA, particularly non-animal forms, remains poorly characterised. This study investigates the stability, absorption, metabolism, and biological effects of a novel fungal-derived HA extracted from Tremella fuciformis using a sequential in vitro multi-barrier model simulating human physiological compartments, including gastric, intestinal, hepatic, renal, chondrocyte, and keratinocyte environments. Across the gastrointestinal stages, fungal-derived HA demonstrated high structural stability, maintained molecular weight, and exerted superior antioxidant and anti-inflammatory activity compared with sodium hyaluronate. It efficiently crossed the intestinal barrier without increasing hyaluronidase activity, indicating protection from premature enzymatic degradation. In hepatic cells, fungal-derived HA exhibited reduced intracellular uptake and greater extracellular persistence, suggesting lower first-pass metabolism and suggesting improved persistence under in vitro conditions. At peripheral targets, it increased the cluster of differentiation 44 (CD44) expression and HA internalisation in chondrocytes and keratinocytes, supporting anti-inflammatory and pro-regenerative effects. Renal assessments revealed minimal excretion and no cytotoxicity, supporting potential systemic availability. Overall, these results provide the first integrated in vitro evidence describing the absorption, distribution, metabolism, and excretion process of fungal-derived HA. This supports the conclusion that this form of HA is stable, biocompatible, and bioactive with therapeutic potential for joint and skin health, as suggested by the in vitro models. Full article

Metabolic and bariatric surgery (MBS) has evolved into a highly effective neurohormonal intervention for severe obesity; however, it introduces unique long-term vulnerabilities, particularly regarding alcohol (AUD) and substance use disorders (SUD). This review synthesizes the epidemiological, pharmacokinetic, and neurobiological drivers of postoperative substance misuse. Procedures like Roux-en-Y gastric bypass (RYGB) radically alter ethanol metabolism, eliminating first-pass metabolism and accelerating gastric emptying, while simultaneously recalibrating reward pathways, creating a “reward gap” that facilitates addiction transfer. These physiological shifts exacerbate critical micronutrient deficiencies (thiamine, B12, iron), increase the risk of post-bariatric hypoglycemia, and correlate with higher rates of liver cirrhosis and suicide. Furthermore, substance use is a primary driver of suboptimal weight loss trajectories and weight regain. Mitigation requires a lifelong, multidisciplinary framework involving preoperative risk stratification, validated screening (e.g., AUDIT-C), and targeted nutritional supplementation to safeguard the long-term metabolic and psychological benefits of MBS. Full article

Biomarker-based guided delivery of drugs is an emerging paradigm of precision medicine in which targeted therapeutic intervention is administered on the basis of certain biological markers in order to achieve maximal dosing, targeting, and time optimization. By utilizing quantifiable physiological or molecular signatures like the expression of transporters, enzymatic activities, metabolite levels, or disease-specific markers to tie in the correlation of drug disposition, these systems provide individualized intervention with optimized efficacy and safety. Oral administration of drugs is still the best route in patient compliance; however, several drugs are handicapped by suboptimal bioavailability secondary to poor solubility, limited permeability, efflux transporter participation, and enzymatic first-pass degradation. These result in variable therapeutic results in patient populations. Biomarker guidance in oral drug delivery provides a potent strategy for overcoming such challenges through site-specific release, real-time dose optimization, and adjustment of absorption pathways. Recent developments include pH-controlled formulations for gut-specific targeting, enzyme-activated nanocarriers, glucose-starved responsive devices for metabolic disease, and biomarker-driven transporters for permeability enhancement. Preclinical and early-phase clinical studies hold promising prospects for applications in oncology, infectious disease, inflammatory bowel disease, and metabolic disease. While promising momentum exists, transition to routine use in the clinic awaits rigorous biomarker validation, scalability in manufacture, and regulations harmonization. On the horizon, the integration of biomarker-guided oral drug delivery with nanotechnology, artificial intelligence, machine learning, and wearable biosensors holds promise for revolutionizing oral therapy into very personalized, responsive, and efficient treatment methods. Full article

Background/Objectives: Current clinical evidence suggests that cannabidiol (CBD) demonstrates therapeutic potential in the management of chronic pain, particularly in conditions involving inflammation. However, its therapeutic potential is severely limited by poor oral bioavailability, extensive first-pass metabolism, and the need for frequent high-dose administration, which compromises patient adherence and tolerability. Long-acting injectable (LAI) delivery systems offer a strategy to overcome these limitations by providing sustained plasma concentrations and reducing dosing frequency. This study aimed to develop and optimise CBD-loaded poly (lactic-co-glycolic acid) (PLGA) microspheres for LAI delivery and to evaluate poly(2-ethyl-2-oxazoline) (POx) as a functional and biocompatible alternative to the conventionally used poly (ethylene glycol) (PEG). Methods: CBD-loaded microspheres were prepared using emulsion–solvent evaporation technique. The formulations were optimised based on entrapment efficiency (EE), drug loading (DL), particle size distribution, surface morphology, thermal behaviour, in vitro release kinetics, and cytocompatibility using NIH 3T3 fibroblasts. Multiple in vitro release methodologies, including dialysis bag, shaking-flask, and USP Apparatus IV, were evaluated to identify the most discriminative and practical approach for long-term release assessment. Results: The optimised POx-based microspheres demonstrated superior control over particle size, yielding significantly smaller and more uniform particles compared with PEG-based microspheres (124 ± 1.47 µm vs. 218 ± 13.5 µm, respectively). Differential scanning calorimetry (DSC) confirmed molecular dispersion of CBD within the polymer matrix. In vitro release studies demonstrated sustained drug release over 20 days. Conclusions: POx represents a promising alternative to PEG for the formulation of CBD-loaded PLGA microspheres, offering enhanced physicochemical stability and biological compatibility. This platform supports the development of safe and effective long-acting injectable CBD therapies and consideration of POx as an alternative to PEG. Full article

The brewing process generates substantial by-products rich in potentially bioactive compounds (e.g., polyphenols and fermentation metabolites), providing a sustainable and appealing source of cosmetic ingredients. Oil-in-water (O/W) emulsions containing 20% (w/w) aqueous extracts from Bionda Triplo Malto beer, wort, and key brewing by-products (hops, yeast, and spent grain) were developed and evaluated using a combined in vitro–in vivo approach. Aqueous extracts were first screened on human immortalized dermal fibroblasts (BJ-5ta) at 0.25–1 mg/mL for cytocompatibility and antioxidant activity. Within this concentration range, no significant changes in cell viability or intracellular antioxidant capacity under UV stress were detected, suggesting cytocompatibility but limited inherent activity. When incorporated into O/W emulsions and tested at an active-equivalent concentration of 10 mg/mL, the formulations increased fibroblast metabolic activity and antioxidant response. In contrast, free extracts at 10 mg/mL showed concentration-dependent cytotoxicity for some matrices, with beer- and yeast-based emulsions demonstrating the strongest effects. The emulsions exhibited good physicochemical stability (pH ~5.7–6.2; viscosity 4750–5150 mPa·s), passed the ISO 11930:2012 challenge test, and were well tolerated in patch testing. In a double-blind, randomized split-forearm study on 50 healthy volunteers over 30 days, beer, yeast, and spent grain-based formulations improved skin parameters versus baseline. TEWL decreased (e.g., beer: 16.22 ± 5.12 to 10.77 ± 2.22 mg·m⁻²·h⁻¹; yeast: 16.29 ± 5.66 to 10.18 ± 1.08; spent grain: 14.45 ± 4.34 to 11.66 ± 2.28), hydration increased (beer: 35.15 ± 5.93 to 42.26 ± 3.78; yeast: 33.27 ± 4.87 to 42.92 ± 2.48; spent grain: 34.22 ± 5.19 to 41.16 ± 3.17, and elasticity improved for beer and yeast formulations (62.33 ± 3.27 to 70.24 ± 2.12 N/m) and yeast (61.21 ± 4.72 to 72.13 ± 5.55 N/m). Based on these findings, brewing-derived ingredients demonstrate potential as cosmetic actives, with formulation critically determining their clinical efficacy. Full article

Insulin injection remains the best therapy for diabetes mellitus, but subcutaneous injection continues to pose challenges, including patient discomfort, poor compliance and fluctuating plasma glucose profiles. Recently, transdermal insulin delivery has emerged as a non-invasive strategy that bypasses gastrointestinal degradation and first-pass hepatic metabolism, thereby increasing insulin bioavailability and enhancing patient acceptance. Recent developments in nanomedicine have facilitated the development of transdermal patches with enhanced drug encapsulation, uptake and controlled release. Nanostructured lipid carriers, polymeric nanocomposites, liposomes and SLNs have demonstrated a five-fold enhancement of transdermal flux and an extended insulin effect in preclinical models. The addition of ionic liquids and polymeric nanogels leads to an additional increase in insulin aqueous solubility and permeation, resulting from the temporary regulation of stratum corneum lipid organization. Bright and stimuli-responsive patches with glucose oxidase or phenylboronic acid functional groups enable regulated insulin delivery in response to changes in blood glucose, demonstrating near-normoglycemia for up to 48 h in animal testing. Nanocomposite systems assisted by microneedles have also been advanced to the early clinical phase, offering enhanced reproducibility of their pharmacokinetics and a low risk of dermal irritation. Despite these encouraging results, several translational challenges remain, such as biocompatibility, repeatability in the production of nanocarriers, long-term stability of formulations and regulatory standardization. This review examines the physicochemical design principles, materials innovations and permeation mechanism of nanomedicine-engineered insulin patches, the current state of preclinical and clinical advancements, challenges in production and future perspectives in viable patient-focused transdermal insulin delivery. Full article

Emodin, a trihydroxy-methyl anthraquinone abundant in rhubarb, Polygonum species, and other medicinal plants, exemplifies the therapeutic potential and translational complexity of the broader anthraquinone scaffold. Anthraquinone derivatives have demonstrated antiproliferative, anti-inflammatory, metabolic, cardiovascular, antifibrotic, and immunomodulatory effects, consistently reported across diverse preclinical models, targeting pathways such as NF-κB, PI3K/AKT, MAPKs, AMPK, PPARs, NLRP3, and ferroptosis-related axes. Despite strong preclinical efficacy, clinical development has been limited by unfavorable absorption, distribution, metabolism, and excretion (ADME) characteristics, including poor aqueous solubility, extensive first-pass glucuronidation, and active efflux via intestinal and hepatic transporters. These features result in low and variable systemic exposure, while high local concentrations, particularly in the gastrointestinal tract, contribute to context-dependent toxicity signals that complicate risk assessment. The present review integrates pharmacological, toxicological, and formulation-focused evidence to provide a unified assessment of emodin and the anthraquinone scaffold. Particular emphasis is placed on bidirectional, dose- and context-dependent effects on the liver and kidney; the modulation of cytochrome P450 enzymes, UGTs, and transporters; and emerging preclinical formulation strategies that aim to decouple intrinsic bioactivity from pharmacokinetic limitations. Full article

The majority of approved drug products comprise formulations of either chemically synthesized small molecules or large molecular entities derived from living cells, commonly referred to as biologics. Over the past two decades, there has been remarkable growth in the approval of biologics for a variety of disorders, including respiratory diseases. The preference for biologics stems from their high target specificity, strong binding affinity, and favorable safety profiles. Most approved biologics are peptides or proteins, which are unsuitable for oral administration due to negligible bioavailability, resulting from their large molecular size, polarity, and susceptibility to enzymatic degradation in the gastrointestinal tract. Consequently, the majority of biologics are administered parenterally, delivering the drug systemically to reach target sites. However, achieving therapeutic concentrations of locally acting respiratory drugs in the lungs via systemic delivery often requires high doses, which increases the risk of adverse effects. For respiratory disorders, nasal and pulmonary drug deliveries are the preferred noninvasive routes. These routes bypass gastrointestinal and first-pass metabolism and deliver therapeutic agents directly to their local site of action. This approach enables a faster onset of action, reduces the required dose by orders of magnitude, and significantly lowers the risk of systemic adverse effects. These advantages have driven the successful development of inhaled formulations for certain rescue and maintenance medications that were originally administered orally or parenterally. Despite this, treatment options for respiratory diseases remain largely limited to small molecules, with only a single inhaled biologic approved in 1993, even though several parenterally administered biologics have since been approved for pulmonary disorders. The scarcity of inhaled biologics is primarily due to the inherent complexity of these drug substances, which impacts all stages of product development, including manufacturing, characterization, purification, stability, formulation design, delivery, and preclinical and clinical evaluations of safety and efficacy. Additionally, sponsors’ interest in developing inhaled biologics may be tempered by the lack of regulatory guidance addressing the multidisciplinary and intricate nature of their development. This article, together with the accompanying review, addresses both regulatory considerations and scientific challenges in the development of inhaled biologics. To the authors’ knowledge, these works represent seminal efforts to examine available regulatory guidance and the applicable literature across various phases of product development beyond safety and efficacy evaluations. We examined the formal regulatory expectations and summarized the requirements as they apply to inhaled products and inhaled biologic protein therapeutics. In parallel, we explored scientifically relevant considerations in the development of inhalation-specific protein therapeutics for which regulatory guidance remains limited, evolving, or absent. While they should not be considered definitive, it is hoped that these contributions will stimulate scientific and regulatory interest, ultimately promoting the identification and resolution of gaps to advance the development of locally acting biologics and address unmet patient needs. Full article

This study aimed to develop dissolving microneedles (MNs) for the buccal delivery of cannabidiol (CBD). CBD is a non-psychotomimetic phytocannabinoid with anti-inflammatory and anxiolytic properties. The MN arrays were produced using micromolding, which has the ability of scalability. However, this approach lacks the ability to customize needle geometry; thus, additive manufacturing was implemented in the study. Digital Light Processing (DLP) printing is a promising way to produce molds with customized MN architecture. In the present study, molds were fabricated from 3D-printed MN arrays to prepare dissolving MNs for buccal administration. Polymeric needles based on Eudragit L100-55 and Eudragit RSPO were produced from reverse molds and they were evaluated regarding their physiochemical and mechanical properties, followed by in vitro and ex vivo studies using porcine buccal mucosa. Visualization studies were conducted using confocal scanning laser microscopy, whereas the membrane integrity of the porcine mucosa upon application of the MNs was assessed by histological evaluation. Our results suggest that the needles can be effectively inserted into the buccal tissue and release the active pharmaceutical ingredient (API) in a controlled manner. This approach offers a patient-friendly alternative to oral CBD delivery, bypassing first-pass metabolism. Full article

Controlled drug delivery systems (CDDSs) are increasingly attracting interest from the scientific community in order to achieve highly precise, customized, and efficient therapeutic treatment of various diseases. The challenge is to develop highly innovative devices and appropriate administration methods in order to reduce side effects and further improve patient compliance. In this context, transdermal drug delivery systems (TDDSs) represent smart tools that permit supplying therapeutically effective amounts of drugs at a fixed time using the skin as the administration route. They are non-invasive and allow for avoiding gastric side effects and first-pass metabolism occurring in the liver. TDDSs have been produced using numerous therapeutic agents and, more recently, also biological molecules. However, it must be highlighted that they are complex systems, and their formulation requires a multidisciplinary approach and expertise in polymer chemistry and materials science. A contribution in this direction is given from the integration of membrane technology with biological and pharmaceutical sciences. The present review deals with a general overview of controlled drug delivery systems. Particular attention is devoted to TDDSs and to the materials used for producing polymeric membrane-based TDDSs with a membrane engineering perspective. It also describes the passive and the most advanced active strategies for transdermal delivery. Finally, different transdermal membrane-based release systems, like patches, mixed-matrix membranes, and imprinted membranes are discussed. Full article