Natural polysaccharide-based multiparticulate systems are promising alternatives to synthetic polymers for oral modified-release applications due to their biocompatibility, low toxicity, and tunable network properties, particularly for drugs associated with gastrointestinal irritation, such as aceclofenac. Here, κ-carrageenan/agar beads were prepared by ionotropic gelation and modified with proanthocyanidin (PA) and tannic acid (TA) as physical crosslinking agents, or carboxymethylcellulose (CMC) as a release-modifying agent. Their effects on drug entrapment efficiency, gastro-resistance, and in vitro aceclofenac release were evaluated. All formulations showed entrapment efficiencies ranging from 78.8 to 92.9%, although lower values were observed for TA-containing beads, likely due to pH-dependent effects. In simulated gastric fluid, PA-modified beads exceeded the predefined 10% release threshold adopted from the USP criterion for delayed-release dosage forms, whereas CMC improved gastric-phase drug retention, with CMC2 limiting aceclofenac release to 8.4%. Selected CMC-modified beads exhibited concentration-dependent swelling and sustained release in simulated intestinal fluid, with a significantly slower release profile than the commercial reference formulation. Kinetic modeling suggested release governed by diffusion coupled with polymer relaxation and/or surface erosion. Physicochemical characterization indicated that CMC promoted structural changes consistent with a more restrictive polymer network, resulting in reduced aceclofenac release under simulated gastric conditions and prolonged release at intestinal pH. Overall, CMC effectively modulated the structural and release properties of κ-carrageenan/agar beads, supporting their potential as a natural polymer-based platform for gastric-phase drug retention and controlled oral delivery of aceclofenac.
T cell receptor-engineered T cell (TCR-T) therapy holds promise for solid tumors but faces challenges from insufficient and non-uniform antigen presentation and inefficient T-cell tumor delivery. Current strategies that rely on endogenous tumor antigen expression are often limited by transcriptional silencing or downregulation of target antigens in a substantial fraction of tumor cells, resulting in incomplete tumor recognition. To address this, we developed iRGD-modified liposomal nanoparticles (iRNY-LNPs) as a universal antigen-delivering platform for remodeling tumors and enhancing TCR-T immunotherapy. The iRNY-LNPs, co-assembled from DSPE-PEG-iRGD and DSPE-PEG–NY-ESO-1 (New York Esophageal Squamous Cell Carcinoma-1), exhibited a uniform size of 206.3 ± 5.4 nm and demonstrated significantly enhanced penetration into 3D tumor spheroids compared to non-targeted nanoparticles. In vitro, iRNY-LNPs enabled uniform NY-ESO-1 antigen presentation on tumor cells, potently activating TCR-T cells and enhancing interferon-γ secretion and cytotoxicity. In vivo imaging confirmed efficient tumor accumulation and retention of iRNY-LNPs. In a gastric cancer model, the combination of iRNY-LNPs and NY-ESO-1 TCR-T cells achieved 73.0% tumor growth inhibition and significantly prolonged survival, with no observed systemic toxicity. This study establishes a proactive tumor-remodeling strategy that circumvents the reliance on endogenous antigen expression by enforcing uniform antigen display. Given its modular design based on lipid–peptide conjugates, the platform can be readily adapted to deliver other tumor-associated antigens by simply substituting the peptide moiety, offering a versatile framework for TCR-T immunotherapy against diverse solid tumors.
A key challenge in the development of localized intestinal enzyme systems is to retain enzyme carriers at the mucosal interface while preserving their catalytic functionality. To our knowledge, retention of enzyme-loaded layer-by-layer polyelectrolyte microcapsules (PMCs) on native small-intestinal mucosa has not previously been demonstrated. In this study, we investigated the ex vivo retention of β-galactosidase-loaded PMCs on porcine small-intestinal mucosa and determined whether the retained capsules preserved catalytic activity. FITC-labelled PMCs were incubated with mucosal explants from the duodenum, jejunum, and ileum and subjected to a standardized washout procedure. Capsule retention was quantified fluorometrically, while PMC-associated surface coverage was assessed by fluorescence microscopy. PMCs showed high retention across all intestinal segments, with a mean retained fraction of approximately 88% and an apparent mucosal surface coverage of approximately 32%. Within the tested conditions, retention was not significantly affected by nominal capsule diameter (1–10 μm), the sacrificial template used for capsule preparation (CaCO3 or MnCO3), NaCl concentration (0–1 M), or Triton X-100 treatment, whereas a significant decrease was observed at pH 4.0. Importantly, PMCs retained on the mucosal tissue remained catalytically active, demonstrating preservation of β-galactosidase functionality after tissue association and washout. These findings provide the first ex vivo demonstration that enzyme-loaded LbL polyelectrolyte microcapsules can be retained on native small-intestinal mucosal tissue while remaining catalytically functional and establish a basis for their further development as mucosa-associated intestinal biocatalytic systems.
