Poly(ethylene glycol) (PEG)-lipids endow lipid nanoparticles (LNPs) with colloidal stability but elicit anti-PEG immunity upon repeat dosing. Here it is replaced PEG-lipids with microbial rhamnolipids (RLs) to create PEG-free, self-adjuvanting LNPs. RL-LNPs show higher than 90% mRNA encapsulation and enhanced dendritic cell uptake via the mannose receptor and DEC205, redirecting expression to lymph nodes after intramuscular injection. Compared with size-matched PEG-LNPs, RL-LNPs enhance transfection and neutralizing capacity against pseudoviruses, while potently amplifying Th1-biased humoral and cytotoxic T-cell responses. Quantitative proteomics and western blotting reveal activation of the C-type lectin receptor (CLR)/NF-kappa B axis, corroborating intrinsic adjuvancy. Notably, RL-LNPs avoid PEG-associated mast-cell infiltration under repeat dosing while maintaining favorable systemic chemistry panels and histology. These data establish glycolipids as dual-function substitutes for PEG-lipids, coupling stabilization with receptor-programmed immunity for next-generation mRNA vaccines.

Successful surgical resection of solid tumours requires highly reliable real-time intraoperative tools to accurately delineate tumour boundaries, which remains challenging in routine clinical standards. Here, we identify endogenous substances with intense autofluorescence in the second near-infrared window (NIR-II, 1,000–1,700 nm) that are abundant in human liver tissues but negligible in cancerous tissues. Inspired by this discovery, we develop a label-free and wide-field imaging approach, named tissue autofluorescence NIR-II imaging (TANI) for visualizing human liver malignancies. TANI demonstrates exceptional contrast (7.69 ± 0.52), sensitivity (97.8

Mucosal immunity-the body's frontline defense, harboring 80% of the body's immune cells-represents a potent yet underexploited avenue for cancer vaccination. Here, we developed an oral biohybrid vaccine platform by integrating tumor antigen-loaded liposomes with fimbriae-enriched bacteria (Escherichia coli or VNP20009) through bacterial hitchhiking or membrane hybridization. These biohybrids promote mucosal antigen delivery via glycoprotein 2 (GP2)-mediated microfold-cell (M-cell) transcytosis, enhancing antigen cross-presentation and activation of a mucosa-periphery-tumor immune cascade. Bacterial membrane-hybridized vaccines outperform bacteria-hitchhiking counterparts by reconfiguring dendritic cell (DC) subsets within gut-associated lymphoid tissues (GALTs) and triggering C-C chemokine receptor type 7 (CCR7)-dependent immune cell trafficking, thereby propagating mucosal immune activation toward distal tumor microenvironment (TME) reprogramming and tumor control. When combined with PD-1 blockade, this strategy enhances antitumor efficacy by promoting effector cell mobilization and establishing memory against tumor rechallenge. Collectively, these findings position bacteria-derived arsenal biohybrids as a versatile oral vaccine strategy, advancing mucosal immunotherapy for cancer.

Lipid nanoparticles (LNPs) are essential for efficient messenger RNA (mRNA) delivery but induce innate inflammation, exhibit limited lymphatic transport, and fail to stimulate adaptive immunity robustly. To address these challenges, we developed quercetin-glucoside derivatives (QG) and introduced them into the LNP system. Among the synthesized derivatives, QG2 (quercetin-di-glucoside) with a 30% molar substitution, maintained optimal nanoparticle stability and enhanced mRNA transfection efficiency in vivo. QG2-LNPs showed increased lymph node transfection, improved dendritic cell activation, and significantly higher SARS-CoV-2 mRNA-driven humoral and cellular immune responses compared to conventional LNPs. Notably, QG2-LNPs substantially reduced local neutrophil infiltration and systemic proinflammatory cytokines while maintaining excellent biocompatibility. These findings highlight quercetin-glucoside as a promising modular design for LNPs that enhances delivery efficiency while reducing inflammatory responses.

Oral drug administration, preferred for patient compliance and convenience, encounters significant bioavailability challenges arising from complex physicochemical, physiological, and formulation-related barriers. Traditional models for evaluating absorption exhibit limitations in physiological relevance and predictive accuracy. This review critically examines evolving methodologies for oral drug absorption assessment, from classical permeability assays to emerging predictive platforms, emphasizing integration of mechanistic research with predictive modeling to advance intelligent platforms and clinical translation. While classical permeability models including parallel artificial membrane permeability assay (PAMPA), Caco-2, Ussing chambers, and intestinal perfusion remain indispensable for early screening, their limited physiological relevance and inability to elucidate mechanistic pathways constrain predictive accuracy. Mechanistic approaches such as lymphatic transport studies and advanced tracking techniques using fluorescence, Single Photon Emission Computed Tomography (SPECT), or Positron Emission Tomography (PET) imaging reveal critical biological pathways but suffer from scalability challenges. Emerging platforms provide transformative potential, as organoid-on-a-chip offer physiologically relevant and high-throughput alternatives, while integrated computational modeling enables multiscale prediction, from molecular interactions to systemic pharmacokinetics. Future oral absorption assessment requires converging in vitro biomimetics, computational modeling, and Artificial intelligence (AI) within unified platforms. Achieving this requires standardized in vitro models, robust data integration, and alignment with regulatory to accelerate translational drug development.

Peptides are increasingly favored as therapeutic agents due to their high efficacy, selectivity, and minimal side effects. However, they often face challenges related to poor stability and limited permeability through the gastrointestinal tract (GIT) and epithelia, necessitating parenteral administration. Despite this, there is a considerable demand for oral administration in clinical practice. To address the urgent clinical need for oral delivery, researchers have developed various technologies to surmount these challenges, including device-related systems, permeation enhancers (PEs), nanocarrier-based systems, and more. This review systematically explores the physiological barriers impacting peptide permeability and discusses the permeation-enhancing technologies designed to overcome them. It also reviews the oral peptide delivery systems currently available or under clinical investigation, offering insights into future developments in this field.

Outer membrane vesicles (OMVs) are nanoscale lipid-bilayer vesicles naturally released by Gram-negative bacteria. By packaging membrane proteins, lipopolysaccharide-derived pathogen-associated molecular patterns, nucleic acids, and metabolites, OMVs integrate intrinsic bioactivity with cargo capacity and have emerged as a versatile platform for therapeutic delivery and immune modulation. In this review, we summarize current understanding of OMV biogenesis, composition, and physicochemical features that shape biodistribution and immunogenicity. We then discuss engineering strategies ranging from genetic rewiring of parental strains and detoxification of lipid A to surface functionalization, hybrid membrane assembly, and stimuli-responsive formulations that enable controllable targeting, loading, and release. Recent progress is highlighted in anti-tumor therapy, nanovaccines for infectious diseases and cancer, and nucleic acid delivery for gene regulation. Finally, we analyze key barriers to clinical translation, including endotoxin-associated safety, batch-to-batch heterogeneity, scalable manufacturing, and standardization of quality attributes. Addressing these challenges through rational design and robust production pipelines will be essential to advance OMV-based therapeutics toward safe, effective, and manufacturable clinical products.

Plant-derived extracellular vesicles (PDEVs) are nanoscale particles isolated from plant tissues that carry proteins, lipids, nucleic acids and secondary metabolites, and function as mediators of intercellular communication. PDEVs have been increasingly investigated in both basic research and translational medicine. Owing to their intrinsic anticancer activity, favorable biocompatibility, and capacity for cargo association/loading and delivery, PDEVs are being explored as nanotherapeutics and drug delivery systems for cancer treatment. This review summarizes the biological features of PDEVs, their mechanisms of action in cancer therapy, and current strategies for engineering multifunctional PDEV-based therapeutic platforms. Finally, the review discusses their potential as drug delivery platforms and analyzes current strategies, advantages, and challenges related to clinical translation. This review aims to advance understanding of PDEV biology and support their future clinical development.

D100B is a first-in-class small-molecule activator of AMP-activated protein kinase (AMPK) that specifically targets the lysosomal pool, enabling precise metabolic regulation with lower systemic toxicity. Despite its favorable solubility and stability, D100B exhibits extremely poor oral bioavailability due to strong mucoadhesion and extensive retention within the intestinal mucus. Electrostatic and hydrophobic interactions between D100B and mucin were found to severely hinder its diffusion and epithelial absorption. To overcome this limitation, a PEGylated self-nanoemulsifying system (PSNE) was developed to reduce mucin binding and enhance mucus penetration. The optimized PSNE displayed uniform nanoscale droplets, sustained drug release, and significantly improved diffusion in simulated mucus. In Caco-2/HT29-MTX co-culture monolayers, PSNE significantly enhanced epithelial transport, while pharmacokinetic evaluation demonstrated a 2.66-fold increase in oral bioavailability compared with the unformulated drug. Overall, this study establishes a mucus-barrier-focused formulation strategy that may be applicable for improving the oral delivery of amphiphilic compounds whose absorption is compromised by mucus-mediated retention.

Oral delivery of peptide therapeutics remains challenging due to gastrointestinal degradation, poor epithelial permeability, and extremely low bioavailability. To address these limitations, we developed an enteric solid formulation based on sorbic acid-choline ionic liquids (ILs) for the oral delivery of semaglutide (Sema), a glucagon-like peptide-1 (GLP-1) analogue. The IL-based enteric system was designed to enhance peptide stability, reduce gastric degradation, and promote intestinal absorption. In vitro studies demonstrated strong resistance to acidic conditions and pH-responsive release in simulated intestinal fluid. In vivo imaging further revealed prolonged intestinal retention of the IL-loaded enteric particles. Pharmacokinetic evaluation showed a 2.3-fold increase in maximum plasma concentration compared to the reference Rybelsus. In type 2 diabetes mellitus (T2DM) mice, the formulation achieved glucose-lowering efficacy comparable to subcutaneous Sema administration, with additional improvements in hepatic histology. Importantly, repeated-dose studies indicated favorable systemic and gastrointestinal tolerability under the tested conditions. Collectively, these results demonstrate that IL-based enteric formulation enhances oral peptide exposure while maintaining safety, offering a promising strategy for noninvasive T2DM management.

Elucidating in vivo lipolysis is crucial for clarifying the underlying mechanisms and in vivo fates of lipid-based nanocarriers, which are essential oral drug delivery carriers. Current mainstream methodologies use various in vitro digestion models to predict the in vivo performance of lipid formulations; however, their accuracy is often impeded by the complicated environment of the gastrointestinal tract. Although fluorescence labeling with conventional probes partly reveals the in vivo translocation of lipid nanocarriers, it fails to elucidate the lipolysis process because of poor signal discrimination among nanocarriers, free probes, and mixed micelles (lipolysis end-products). Here, a polarity-sensitive probe (PN-C18) with aggregation-caused quenching properties for labeling lipid nanocarriers is developed and optimized. PN-C18 successfully eliminates interference from both free probes and mixed micelles during lipolysis. In a representative in vitro lipolysis model, PN-C18 labeling shows stronger correlation between fluorescence intensity and lipolysis progression than those of previous methods. In vivo, the translocation and lipolysis of lipid nanoparticles are clearly visualized and effectively monitored, owing to the high tissue-penetrating capability of PN-C18 NIR-II photons. This study provides practical means for elucidating the in vivo fate of lipid-based drug delivery systems and offers valuable insights and reference for further studies in this domain.

Profiling in vivo release kinetics of drug nanocarriers is of high translational significance. However, this has remained unrealized due to the lack of direct methodologies to quantify either the total released or residual drugs. This study employed an indirect strategy, comparing pharmacokinetics and particokinetics, to estimate the in vivo release kinetics of paclitaxel (PTX) from intravenously administered mPEG-PDLLA polymeric micelles (PMs). Blood pharmacokinetics were profiled by chromatographically quantifying PTX, while particokinetics were determined following labeling PM particles by near-infrared fluorophores with aggregation-caused quenching properties. By monitoring the dynamic change in the PTX-to-copolymer ratio, the in vivo release of PTX from the PMs was estimated. The results revealed surprisingly rapid release, with over 88.2% and 99.0% of PTX released by 15 s and 5 min post-administration, respectively. It is concluded that PTX is released rapidly from PMs in vivo, and PMs may merely work as “solvents” to solubilize PTX rather than as carriers for targeted delivery.

Modulation of gut microbiota has emerged as a promising therapeutic strategy for inflammatory bowel disease (IBD). However, current interventions such as probiotics and fecal microbiota transplantation remain limited by insufficient safety and efficacy. To address this, we engineered commensal Lactobacillus rhamnosus (LGG) using miRNA-loaded biomimetic nanoparticles to enhance its proliferation and indole-3-carboxaldehyde production. By functionalizing bacterial extracellular vesicles (BEVs) derived from LGG with lipid nanoparticles (LNPs), we developed BEV-LNPs that exhibited enhanced targeting efficiency toward LGG compared to Escherichia coli. In vitro and in vivo studies demonstrated that BEV-LNPs showed superior stability in simulated physiological fluids and gastrointestinal environments compared to conventional LNPs. When combined with 5-aminosalicylic acid, the BEV-LNP formulation notably improved outcomes in acute and chronic colitis models, reducing inflammation, restoring epithelial barrier integrity, and promoting microbial balance. This study presents an effective strategy for colitis treatment by leveraging miRNA-loaded nanoparticles.

Inflammatory bowel disease (IBD) is a chronic and recurrent inflammatory disease that affects the gastrointestinal tract. The major hurdles impeding IBD treatment are the low targeting efficiency and short retention time of drugs in IBD sites. Nanoparticles with specific shapes have demonstrated the ability to improve mucus retention and cellular uptake. Herein, mesoporous silica nanoparticles (MSNs) with various morphologies were used to deliver budesonide (BUD) for the treatment of IBD. The therapeutic efficacy is strongly dependent on their shapes. The system comprises different shapes of MSNs as carriers for budesonide (BUD), along with Eudragit S100 as the enteric release shell. The encapsulation of Eudragit S100 not only improved the stability of MSNs-BUD in the gastrointestinal tract but also conferred pH-responsive drug release properties. Then, MSNs efficiently deliver BUD to the colon site, and the special shape of MSNs plays a critical role in enhancing their permeability and retention in the mucus layer. Among them, dendritic MSNs (MSND) effectively reduced myeloperoxidase (MPO) activity and levels of inflammatory cytokines in the colon due to long retention time and rapid release in IBD sites, thereby enhancing the therapeutic efficacy against colitis. Given the special shapes of MSNs and pH-responsivity of Eudragit S100, BUD loaded in the voids of MSND (E@MSNs-BUD) could penetrate the mucous layer and be accurately delivered to the colon with minor side effects. This system is expected to complement current treatment strategies for the IBD..

Intraperitoneal (i.p.) administered nanomedicine has been widely applied in the clinical treatment of intra-abdominal diseases and preclinical pharmacological investigations. However, current understandings about the in vivo fate of i.p.-administered drug remains controversial owing to lack of reliable investigation tools. This work presents a nanoparticle-labeling strategy based on aggregation-caused quenching (ACQ) probes in the second near-infrared (NIR-II) window, which can eliminate the interference of unbound probes and allow for non-invasive tracking of nanoparticles in deep tissues. Our results strongly evidence a size-dependent absorption and biodistribution of the i.p.-administered polymeric nanocarriers (PNs) with particle sizes ranging from 30 to 1000 nm both in vivo and ex vivo, and moreover provide a clear visualization of lymphatic transportation and lymph node retention of integral PNs. Importantly, our findings suggest that small particles (≤30 nm) are favorable in systemic therapies due to their rapid absorption and high concentration (>19 %ID mL-1) in circulation, while large particles (over 1000 nm) are meant for localized treatment of abdominal diseases. Besides, the high retention of 200 nm nanoparticles within lymph nodes indicates their promising role in cancer vaccines and lymphatic diseases including lymph node metastasis.

Oral delivery of the hydrophobic tretinoin (Tre) is a significant challenge due to poor solubility and bioavailability. Herein, we describe the potential of ionic liquids (ILs, e.g., [Ch][Ger]), in particular active pharmaceutical ingredient-based ILs (API-ILs, e.g., [Ch][Tre]) and their mixtures (e.g., [Ch][Tre]-[Ch][Ger]), for oral delivery of Tre. [Ch][Tre] provided excellent apparent content of Tre (42.10 g/mL), a 175 million-fold improvement over its water solubility. The cumulative Tre release at 5 min of Tre-[Ch][Ger] and [Ch][Tre]-[Ch][Ger] reached 95% and 97%, respectively. Upon the same oral dosing of Tre in rats, [Ch][Tre]-[Ch][Ger] increased peak blood concentration of Tre by 2.9-fold and total Tre exposure by 3.6-fold. Furthermore, the oral bioavailability of Tre in [Ch][Tre]-[Ch][Ger] was significantly enhanced to 361.4%. It's speculated that the mixture of ILs gave rise to sophisticated solvent structures, making the system more stable to enhance the drug adsorption. In conclusion, our findings highlight the potential of ILs technology as a promising platform for oral delivery of hydrophobic drugs.

In polarized cells, the differential distribution of proteins results in the formation of apical and basolateral membranes. The basolateral membrane contacts basal lamina and mediates cell-to-cell communication, which is crucial for maintaining homeostasis and enabling drug absorption. To establish and maintain the basolateral domain, intricate mechanisms are necessary to ensure the proper sorting and transportation of molecules. Sorting signals play a crucial role in regulating the distributions of basolateral proteins, determining their trafficking route and final residence. Newly synthesized proteins can be segregated into different carrier vesicles at either trans-Golgi network (TGN) or endosomes. Additionally, understanding basolateral transport in polarized epithelial cells is important for predicting diseases and delivering drugs. This review provides a summary of recent advancements in the mechanisms and applications of basolateral sorting and trafficking.

Amphiphiles,including surfactants,have emerged as indis-pensable elements in materials science and pharmaceutical sci-ence,and their functions are highly relying on the critical micelle concentration(CMC)[1,2].Numerous fluorimetry-based probes have been developed to measure CMCs[3](Fig.S1).However,CMC measurements using these probes suffer from a time-consuming and laborious procedure and large uncertainties,primarily due to their poor photo-stabilities and highly fluctuating fluorescence backgrounds.

Lipid nanoparticles (LNPs) are currently the most commonly used non-viral gene delivery system. Their physiochemical attributes, encompassing size, charge and surface modifications, significantly affect their behaviors both in vivo and in vitro. Nevertheless, the effects of these properties on the transfection and distribution of LNPs after intramuscular injection remain elusive. In this study, LNPs with varying sizes, lipid-based charges and PEGylated lipids were formulated to study their transfection and in vivo distribution. Luciferase mRNA (mLuc) was entraped in LNPs as a model nucleic acid molecule. Results indicated that smaller-sized LNPs and those with neutral potential presented superior transfection efficiency after intramuscular injection. Surprisingly, the sizes and charges did not exert a notable influence on the in vivo distribution of the LNPs. Furthermore, PEGylated lipids with shorter acyl chains contributed to enhanced transfection efficiency due to their superior cellular uptake and lysosomal escape capabilities. Notably, the mechanisms underlying cellular uptake differed among LNPs containing various types of PEGylated lipids, which was primarily attributed to the length of their acyl chain. Together, these insights underscore the pivotal role of nanoparticle characteristics and PEGylated lipids in the intramuscular route. This study not only fills crucial knowledge gaps but also provides significant directions for the effective delivery of mRNA via LNPs.

Microfluidic technology has emerged as a prevalent tool to produce lipid nanoparticles (LNPs) for nucleic acid delivery. However, its wide-ranging application is hindered by specialized, costly equipment and consumables. Herein, a ready-to-use lipid nanoparticle (RULNP) technology employing deep eutectic solvents (DESs) was developed. The DES, consisting of fructose and glycerol ([Fru][Gly]), was able to dissolve lipids and nucleic acids, facilitating the formation of RULNPs by simple physical mixing and hydrating. This innovative approach circumvents the high costs and organic solvents associated with microfluidic methods and offers flexibility in preparation techniques, accommodating various application scenarios. RULNPs exhibited physicochemical properties and plasmid DNA (pDNA) or RNA delivery efficacy comparable to those of LNPs. Mechanistic studies revealed that RULNPs achieved superior cellular uptake compared with LNPs despite exhibiting limited endosomal escape capabilities. Collectively, the DES-based RULNP system presents a rapid and straightforward method for LNP production, potentially revolutionizing nucleic acid delivery.