Aug 2026· International Journal of Pharmaceutics: X· Vol 12· 0 citations· 78 references
Medicine
TL;DR
Comparing two cationic reverse-hexagonal LCNs formulated from monoolein, oleic acid, poly(allylamine hydrochloride) and either poloxamer 407 (P407) or poloxamer 188 (P188) reveals a direct relationship between LCN composition, membrane affinity, and functional performance, providing mechanistic insight for the rational design of LCN-based carriers in RNA therapeutics.
Abstract
The clinical translation of small interfering RNA (siRNA) therapeutics critically depends on delivery systems capable of protecting nucleic acids, enabling efficient cellular uptake, and promoting cytosolic release. Liquid crystalline nanoparticles (LCNs) are promising carriers due to tunable internal nanostructure and biocompatibility, yet how composition controls membrane affinity and biological performance remains limited. Here, we compared two cationic reverse-hexagonal LCNs formulated from monoolein, oleic acid, poly(allylamine hydrochloride) and either poloxamer 407 (P407) or poloxamer 188 (P188). Both LCN-P407 and LCN-P188 shared reverse-hexagonal organization, mean diameters of 150–185 nm, low polydispersity (0.09–0.18), positive zeta potentials (10–20 mV), and protection of siRNA from RNase A. Langmuir isotherms and Brewster angle microscopy revealed pronounced adsorption and expansion of DPPC monolayers, with LCN-P188 inducing stronger perturbations, consistent with its higher cytotoxicity (∼20% increase in cell death) relative to LCN-P407. Crucially, siRNA delivered by LCN-P407 showed significantly greater uptake (> 1.7-fold) than that administered by LCN-P188. Functionally, as a proof of concept, we showed that LCN-P407-siTNFα induced robust gene silencing in LPS-stimulated macrophages, reducing TNFα secretion by 1.2–3.5-fold depending on particle concentration and incubation time. LCN-P188-siTNFα produced only delayed and modest reductions (1.3–1.6-fold). In addition, in dermatomized porcine skin, LCN-P407 also exhibited superior cutaneous penetration, delivering siRNA efficiently into the viable epidermis and dermis. Collectively, these results reveal a direct relationship between LCN composition, membrane affinity, and functional performance, providing mechanistic insight for the rational design of LCN-based carriers in RNA therapeutics.
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
ABSTRACT Lyotropic liquid crystalline nanoparticles (LCNPs), including cubosomes, are increasingly investigated as antimicrobial nanomaterials because non‐lamellar lipid nanoparticles can fuse with biological membranes, exchange lipids, and improve antimicrobial delivery or antibiotic combination treatment. However, prior studies have mainly addressed fusion, uptake, encapsulation, or payload stabilization, rather than testing whether retained internal curvature can be isolated as a design variable for antibacterial potentiation in a matched LCNP series. Herein, we generated lamellar vesicles, primitive cubosomes (P‐cubosomes, Im3m), and diamond cubosomes (D‐cubosomes, Pn3m) from the same phytantriol/DPPS lipid system. When combined with free daptomycin, rather than being used as drug‐loaded carriers, these LCNPs exhibited curvature‐dependent potentiation hierarchy against methicillin‐resistant Staphylococcus aureus (MRSA), vesicles < P‐cubosomes < D‐cubosomes. Fluorescence imaging, electron microscopy, and neutron reflectometry showed progressively stronger membrane association, lipid extraction, and bilayer disruption with increasingly negative curvature. In a murine bacteremia model using a sub‐optimal daptomycin regimen, the same curvature‐dependent efficacy trend was retained in vivo, providing proof‐of‐concept support rather than therapeutic validation. This study provides direct experimental evidence, in a matched antibacterial LCNP system, that retained internal curvature modulates membrane remodeling and potentiates daptomycin against MRSA.
Xiangfeng Lai, Shuhong Wang, Chenguang Ding et al.· Advancement of science· 0 citations
Understanding the physicochemical factors that govern siRNA nanocarrier assembly is essential for the rational design of effective delivery systems. By optimizing various lipid compositions, cholesterol content and PEG length we created a peptide-functionalized cationic liposomal platform made of DOPE/TAP lipids with cholesterol-anchored nona-arginine (R9-Chol) for siRNA complexation, intracellular transport and effective silencing of the target EGFR gene. Analysis of ζ-potential and dynamic light scattering allowed to rationally design formulation of stable, monodisperse nanoscale lipoplexes with a positive surface charge. With fluorescence polarization, circular dichroism and agarose gel electrophoresis we found an optimal siRNA:liposome complexation ratio of 1:77, which protected siRNA from ribonuclease-mediated degradation. Morphological imaging confirmed a shift from discrete vesicular structures to organized multilamellar lipoplexes, consistent with electrostatically driven self-assembly. In cellular studies, the optimized nanocarrier promoted efficient uptake of fluorescent siRNA in MDA-MB-231 cells and achieved functional delivery of anti-EGFR, leading to substantially reduced expression of the target gene at both transcript and protein levels. This work offers mechanistic understanding of peptide-assisted lipid–siRNA assembly and positions R9-functionalized DOPE/TAP liposomes as a promising platform for siRNA delivery.
P. Białecki, S. Braccia, T. Makowski et al.· bioRxiv· 0 citations
Extracellular vesicles (EVs) have garnered significant interest as potential bioderived drug delivery systems for RNA therapeutics owing to their distinctive characteristics, which include their inherent capability to transport various biological molecules throughout the body. However, several issues still pose challenges, such as the limited loading of exogenous RNAs and poor scalability of conventional techniques. High-pressure homogenization (HPH) is a useful alternative for the scalable preparation of EVs encapsulating small-molecule therapeutic agents via a one-step pharmaceutical process. However, whether HPH can be applied to macromolecular drugs, such as small interfering RNA (siRNA), remains unclear. Herein, we report the applicability of using HPH to prepare siRNA-loaded EVs and their subsequent cytoplasmic siRNA delivery. Simultaneous HPH of bovine milk-derived EVs (mEVs) with distearoylphosphatidylethanolamine-polyethylene glycol-polyethyleneimine and cholesterol-conjugated siRNA allowed siRNA loading onto the mEVs. Modification of the mEVs with the tumor-targeting ligand, cyclo(Arg-Gly-Asp-d-Phe-Lys) (cRGD), showed high affinity for a human glioma cell line, resulting in remarkable intracellular siRNA delivery. Moreover, siRNA delivered with the cRGD-modified mEVs showed a higher knockdown efficiency than that of the unmodified mEVs. The present findings suggest, for the first time, that HPH holds potential for the development of siRNA-loaded EV therapeutics that could be useful for cytoplasmic siRNA delivery.
Tatsuya Fukuta, Masato Miyazaki, Taiki Fujimoto et al.· ACS Applied Bio Materials· 0 citations
Purpose: The development of safe and efficient non-viral carriers remains critical for gene therapy. Biocompatible polysaccharides such as dextran are attractive candidates for novel delivery platforms. This study reports the synthesis, characterization, and evaluation of a cationic dextran derivative for pDNA delivery. Methods: Cationic Dextran was synthesized through periodate oxidation and reductive amination with 1,6-Hexamethylenediamine. Its structure was confirmed using ¹H NMR and FT-IR spectroscopy. Nanoparticles, formed via self-assembly with pDNA, were analyzed for size, zeta potential (DLS), and morphology (SEM). Functional properties, including buffering capacity, pDNA condensation (gel retardation assay), DNase I protection, cytotoxicity in NIH3T3 cells (MTT assay), and cellular uptake, were assessed. Results: Nanoparticles showed uniform size (207–245 nm) and strong positive zeta potential (+57.2 to +73.7 mV). The polymers demonstrated a substantial buffering capacity within the endosomal pH range, efficiently condensed pDNA, and provided partial protection against DNase I. Cytotoxicity was low at both 24 and 48 hours. Flow cytometry confirmed high cellular uptake, with over 95% of NIH3T3 cells internalizing polyplexes. Conclusion: These cationic Dextran nanocarriers demonstrate efficient pDNA condensation, low cytotoxicity, and high cellular uptake, making them promising for gene delivery. Although nuclease protection needs optimization, it offers a versatile platform for non-viral gene therapy.
K. Karami, S. S. Uroomiye, K. Derakhshandeh et al.· Advanced Pharmaceutical Bull...· 0 citations
The development of efficient and targeted drug delivery systems remains a significant challenge, particularly for active pharmaceutical ingredients with poor aqueous solubility. Among various nanocarrier systems, Mesoporous Silica Nanoparticles (MSNs) have emerged as promising candidates due to their high surface area, tunable pore size (2-50 nm), thermal stability, and chemical modifiability. This review comprehensively discusses the rationale behind the utilization of MSN as drug delivery systems, focusing on how the type and concentration of surfactants, along with surface functionalization strategies, influence their physicochemical characteristics and pharmacokinetic performance. The synthesis of MSNs typically involves sol-gel processes using silica precursors (e.g., tetraethyl orthosilicate) and surfactants (e.g., cetyl trimethyl ammonium bromide, Pluronic F127), which dictate the morphology, particle size, and pore architecture of the resulting nanoparticles. Furthermore, surface modifications employing functional groups such as polyethylene glycol or pH-responsive polymers enhance biocompatibility, prolong systemic circulation, and enable controlled and site-specific drug release. Evidence from recent studies demonstrates that MSNs significantly improve drug loading efficiency, enhance solubility and bioavailability, and reduce off-target toxicity. Consequently, MSNs represent a highly versatile and modifiable platform with considerable potential for addressing the limitations of conventional drug delivery systems, particularly in oncology and the treatment of chronic diseases.
Ahmad Ainurofiq, Y. Ramadhana, Salma Aqilah Rachmadani et al.· Recent Advances in Drug Deli...· 0 citations
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