It is demonstrated that apo-Fn undergoes controlled disassembly under mild acidic conditions, and efficiently reassembles upon neutralization or urea removal, and efficiently reassembles upon neutralization or urea removal.
Abstract
Ferritin (Fn) nanocages offer significant potential as drug delivery vehicles due to their biocompatibility, well-defined structure, and inherent targeting capabilities. In this study, we isolated Fn from the liver of Esox lucius and prepared its apo-form (apo-Fn) to engineer its reversible disassembly/reassembly for drug encapsulation. We demonstrated that apo-Fn undergoes controlled disassembly under mild acidic conditions (pH 2.0-4.0) or 4 M urea, and efficiently reassembles upon neutralization or urea removal. Thermal treatment below 55 °C also facilitated reversible structural transitions. Based on these properties, three loading strategies were developed. The temperature-gradient method was optimal for hydrophilic drugs (doxorubicin and phenytoin), while the urea-gradient method achieved 42.18% encapsulation for hydrophobic paclitaxel. The resulting formulations showed uniform size, colloidal stability, and minimal leakage at pH 7.4, but exhibited rapid release at acidic pH 5.0. Furthermore, apo-Fn showed high biocompatibility (> 90% cell viability) and exerted intrinsic anti-inflammatory effects by modulating macrophage polarization. This study highlights piscine apo-Fn as a promising platform for targeted drug delivery.
Chemically modified protein-based nanocomplexes are widely used as food-grade carriers, but their molecular mechanisms remain unclear. This study developed Pickering emulsions (PEs) composed of phosphorylated (PLF) or succinylated (SLF) lactoferrin (LF) complexed with luteolin (LUT) via a pH-driven method, integrating experimental and computational approaches. SLF-LUT PEs showed superior physicochemical properties, including smaller droplet sizes, encapsulation efficiency up to 80% under neutral conditions, and enhanced storage stability at pH 8 over 30 days. SLF-LUT also significantly inhibited lipid oxidation, with reduced primary and secondary oxidation products. Multi-ligand docking and 100 ns molecular dynamics simulations revealed that LUT binds multiple flaxseed oil fatty acids within lactoferrin's hydrophobic pocket, positioning it for radical scavenging at the oil-water interface. The LF-LUT complex remained stable in a lipid-rich environment, with key residues (LEU320, PRO251, SER252) critical for stabilization. Network pharmacology further linked these interactions to antioxidant pathways (KEAP1/NRF2, FoxO, AGE-RAGE). These findings highlight the potential of nanocomplex-based oil-in-water PEs for delivering hydrophobic bioactives in food and pharmaceutical applications.
Tianzhu Guan, Ning Li, Yining Feng et al.· npj Science of Food· 0 citations
The delivery of therapeutic short interfering RNA (siRNA) is hindered by biological barriers such as rapid degradation and poor cellular uptake. This study investigates peptide-based nanogels (NGs) as potential delivery platforms using three N-capped tripeptides, 2NapKFF, 2NapFKF, and 2NapFFK, designed to load siRNA via electrostatic interactions. The NGs were formulated through a "top-down approach" from pH-triggered hydrogels (HGs) and stabilized by surfactants. Biophysical characterization revealed that while all tripeptides achieved ∼99% siRNA encapsulation, the position of the lysine residue significantly influenced the stability and mechanical properties of the network. The 2NapFKF system emerged as the most suitable candidate, maintaining a size (∼200 nm) compatible with parenteral administration. Biological assays on Human Embryonic Kidney (HEK293) cells confirmed biocompatibility and cytoplasmic internalization through endocytic pathways. Furthermore, the treated cells maintained normal mitotic activity, indicating no impairment of cell proliferation. These findings demonstrate that lysine-modified tripeptide NGs are safe and effective tools for gene-silencing applications.
Mariangela Rosa, Fin Hallam Stewart, C. Diaferia et al.· Biomaterials Science· 0 citations
Hierarchical liposome-in-liposome (LIL) structures enable sophisticated multi-stage drug delivery, their application is however often limited by complex, time-consuming fabrication and the use of organic solvents. Here we report a simple two-step hydration (TSH) method for fabricating stable LIL architectures in aqueous media within 2 h. This process employs ultrasonication for the formation of structurally robust inner liposomes (IL) and gentle hydration for their subsequent encapsulation within outer liposomes (OL), achieving high-quality dual-spacing configurations. Optical microscopy and differential staining verified the structural integrity and physical encapsulation, demonstrating distinct spatial segregation. The functional efficacy and temperature-sensitive release kinetics of the LIL system were validated through the release theory, confirming that the hierarchical membranes confine encapsulated agents and prevent premature leakage below the phase transition temperature (Tphase) of the OL. The programmable stepwise release was demonstrated through a macroscopic hydrogelation model, where crosslinking was triggered only upon the sequential thermal release of the initiator and accelerator. This was further validated by the independent release of commercial drugs, chlorpheniramine maleate and riboflavin sodium phosphate, as confirmed via HPLC analysis. These results validate the TSH-based LIL platform as a versatile and scalable strategy for developing smart, multi-stage delivery vehicles with environment-responsive release profiles.
Jin Yoo, Suhyeon Jo, Dong Wook Lim et al.· Colloids and Surfaces B: Bio...· 0 citations
Overall, the Eu-S100@TCh/CS microbead system represents a rationally designed colon-targeted delivery platform with potential to improve local therapeutic efficacy and reduce systemic toxicity.
Huma Hameed, Syed Muhammad Ahmad, Shazia Akram Ghumman et al.· RSC Advances· 0 citations
As a highly symmetrical and endogenous protein-based drug delivery system, ferritin nanocage has garnered tremendous attention in nanomedicine due to its exceptional biocompatibility, dynamic self-assembly behavior, and intrinsic receptor-mediated targeting capabilities. With the rapid evolution of bioconjugation chemistry and protein engineering, the functional manipulation of ferritin has transitioned from passive physical encapsulation to precise, site-specific engineering of ferritin architectures. This Viewpoint systematically highlights the multidimensional bioconjugation strategies of ferritin and its relatives. We specifically focus on the biochemical principles driving genetic fusion engineering, classic covalent chemical conjugation, modular bioorthogonal assembly, and the in situ construction of catalytic centers within the inner cavity. These chemistries are not merely incremental refinements. They unlock entirely new therapeutic and diagnostic capabilities, including single-dose tumor-lethal chemotherapy, high-sensitivity multimodal/nuclear imaging, targeted protein degradation, ultrapure nanovaccines, and inflammation intervention. This Viewpoint aims to provide a chemically grounded and forward-looking roadmap for the next generation of ferritin engineering.
Yang Liu, Ying Xiao, Jiuyang He et al.· Bioconjugate chemistry· 0 citations
