Journal of Liposome Research最新文献

The enzymatic degradation of bile acid-modified phospholipids by secretory phospholipase A₂ (sPLA₂) can induce membrane disruption in liposomes. However, the influence of bile acid hydrophilicity on this disruptive effect remains unclear. In this study, we synthesized four lipids by conjugating lithocholic acid (LCA: 3α-OH), chenodeoxycholic acid (CDCA: 3α-OH, 7α-OH) or its derivatives (7k-CDCA: 3α-OH, 7 = O; bk-CDCA: 3 = O, 7 = O) to a phosphocholine backbone. We found that sPLA₂-mediated release of 6-carboxyfluorescein (6-CF) from liposomes was significantly faster with LCA-PC (100% release) than with the more hydrophilic CDCA-, 7k-CDCA-, or bk-CDCA-PC (14-30% release) in 2 hours. Assays of enzymatic degradation rates indicated that the low efficacy of CDCA-PC and 7k-CDCA-PC correlated with their slow hydrolysis by sPLA₂. Although bk-CDCA-PC was degraded at a rate (13.01%) similar to LCA-PC (15.07%) in 2 hours, dialysis experiments revealed that its metabolite (bk-CDCA) readily diffused into the aqueous phase, unlike LCA, which remained anchored to the membrane. This demonstrates that the firm interfacial settlement of the bile acid metabolite is crucial for the membrane-disruptive effect. Our findings establish that increased hydrophilicity of the bile acid moiety attenuates the sPLA₂-induced pore-forming effect, providing a critical guideline for the future design of sPLA₂-responsive liposomal systems.

Osteoporosis increases fracture risk, necessitating safe therapies. Icariin has anti-osteoporosis effects but exhibits poor solubility, short circulation, and lack of bone targeting. We developed tetracycline (TC)/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol) (DSPE-PEG) co-modified liposomes (Icariin-TC-DSPE-PEG-L) to deliver icariin. Formulation variables (phospholipid/cholesterol, phospholipid/icariin, and phospholipid/TC-DSPE-PEG ratios) were optimized using an orthogonal experimental design with particle size and encapsulation efficiency as responses. Optimized liposomes showed small size (157.5 ± 0.27 nm), zeta potential (-24.14 ± 0.25 mV), low PDI (0.149 ± 0.002), high encapsulation efficiency (92.85 ± 0.06%), and drug loading (10.48 ± 0.17%). Hydroxyapatite binding and in vivo fluorescence imaging confirmed enhanced bone affinity and accumulation. Quantitative pharmacokinetic analysis demonstrated prolonged systemic exposure of Icariin-TC-DSPE-PEG-L compared with free icariin and non-targeted liposomes, indicating improved circulation and bioavailability. In osteoporotic rats, the formulation significantly improved bone microstructure, increased bone mass, and regulated bone homeostasis. The TC/PEG co-modified liposomes effectively overcame limitations of icariin's delivery, thus offering a novel therapeutic strategy for osteoporosis. Notwithstanding, long-term toxicity of TC/PEG co-modified liposomes and mechanisms underlying its therapeutic efficacy will be explored in the not-too-distant future.

Nicotinamide mononucleotide (NMN), a potent nicotinamide adenine dinucleotide (NAD⁺) precursor, has demonstrated significant potential in mitigating mitochondrial oxidative stress, alleviating degenerative diseases, and reducing reactive oxygen species (ROS) generation. However, its topical application is hindered by inherent challenges, including high hydrophilicity, poor skin permeability, and limited stability. To address these limitations, we developed elastic cationic liposomal nanogels (EC-Lip-nanogels), a novel delivery system composed of Nap-FF-GHK nanogels and elastic cationic liposomes (EC-Lips). Nap-FF-GHK peptide was synthesized by coupling the self-assembled scaffold Nap-FF with the anti-aging module GHK, which would be self-assembled into cationic nanogels. EC-Lips was constructed by incorporating edge activators (EAs) into cationic liposomes (C-Lips). The NMN-loaded EC-Lip-nanogels were systematically characterized, demonstrating favorable physicochemical properties with an average particle size of 53.2 nm, a zeta potential of +14.0 mV, and a high encapsulation efficiency of 90.2%. Notably, the penetration enhancement achieved by EC-Lips and EC-Lip-nanogels was 22.7% and 16.0%, respectively, compared to free NMN. This study might establish EC-Lip-nanogels as a promising platform for improving the skin permeability of NMN and other hydrophilic bioactive molecules, offering a transformative approach for topical applications.

Ferulic acid (FA) has multiple anti-aging functions but is limited by poor solubility, low bioavailability, and unclear mechanism in skincare. To overcome these challenges, we have developed ferulic acid ethosomes (FA-ES) co-modified with astragaloside IV and ceramide IIIB. Stability of FA-ES was confirmed through physicochemical characterization and molecular dynamics simulations. Comparative studies in zebrafish models showed FA-ES significantly reduced embryonic toxicity and β-galactosidase activity, improved skin hydration, repair capabilities, and antioxidant effects. In C. elegans models, FA-ES increased transdermal absorption efficiency, extended lifespan, enhanced reproductive capacity, reduced lipofuscin accumulation, and improved resistance to oxidative stress and high-temperature conditions. Overall, FA-ES addresses FA's solubility and bioavailability issues and offers superior skin anti-aging effects, making it a promising transdermal formulation platform for cosmetics.

Based on improved thin-film dispersion method with an optimized preparation process, elevated encapsulation efficiency and excellent stability of the single-chamber liposome, lip@MET/CUR, were achieved for co-delivery of metformin (MET) and curcumin (CUR). To increase the volume of the hydration chamber and the specific surface area during lipid membrane formation, Tween-80 and glass microspheres were introduced in the preparation process. On this basis, the optimal process parameters for high encapsulation efficiency were screened and determined by combining the analytic hierarchy process (AHP), entropy weight method (EWM), and Box-Behnken response surface optimization. Eventually, the optimal encapsulation efficiencies for MET and CUR were determined to be 46.4% ± 1.3% and 94.1% ± 1.5%, respectively. The lip@MET/CUR exhibited an average particle size of 150 ± 2.5 nm with uniform particle size and good storage stability. In vitro drug release experiments revealed a significant sustained-release characteristic of lip@MET/CUR. Specifically, the cumulative release rate of MET decreased from 96.8% to 57.4% within the initial 2 h. Results from MTT assays and experiments conducted in tumor-bearing mice further demonstrated that lip@MET/CUR was more effective in inhibiting the growth of HepG2 cells and tumors compared to free CUR or lip@CUR. In summary, our findings suggest that the optimized lip@MET/CUR formulation holds great potential as a candidate for investigating the synergistic effects of CUR and MET in tumor treatment.

Rheumatoid arthritis (RA) is a chronic autoimmune disorder that undergoes joint pain inflammation and stiffness. Current treatments associated with severe side effects and high costs. Alternative treatment, that is, Epigallocatechin gallate (EGCG) a green tea polyphenol that directly targeted RA inflammatory pathways. But it has bitter taste, poor bioavailability, and toxicity issues. This research aimed to address these limitations through complexation of EGCG with phospholipids (EGCG-PC) and loading it into nanostructured lipid carriers (NLCs). The complexation with phospholipid technique has been shown to be a more successful approach. The solvent evaporation approach has been utilized to manufacture drug complex, which increases the stability and effectiveness of the products. EGCG-PC-NLCs improve solubility, sustained release, and increased bioavailability. The particle size, zeta potential, and PDI of EGCG-PC-NLCs were 159.65 ± 1.34 nm, -21.5 ± 0.99 mV, and 0.148 ± 0.045. It showed sustained drug release in 24 hours where pure EGCG degraded within 4 hours. Anti-rheumatic efficacy was done through in vitro cell viability assays. Toxicity, ex vivo, and in vivo biodistribution studies improved intestinal permeability, exhibited lower toxicity, undergoes lymphatic pathway and avoided rapid clearance. The dual formulation of EGCG phospholipid complex and NLCs showed a safer therapeutic option for RA treatment.

The design of vehicles for transdermal gene delivery is at the forefront of molecular medicine, facilitating targeted therapies. Reports suggest that flexible liposomes can be a good alternative for transdermal delivery, and asymmetric liposomes may enhance gene delivery efficiency. This study aims to create flexible asymmetric-type liposomes with high encapsulation of DNA and high deformability rates. The synthesis of asymmetric liposomes was standardized using the inverse emulsion method, with lipids DOTMA (1,2-di-O-octadecenyl-3-trimethylammonium propane) and DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) as the inner layer, DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine) lipid as the outer layer, cholesterol as a stabilizing component, and Span 80 and ethanol as components that promote flexibility. The pIRES2-EGFP plasmid was used as the encapsulated genetic material. Asymmetric liposomes were characterized using transmission electron microscopy (TEM), encapsulation efficiency percentage (%EE), and the deformability index determined by the extrusion method. Results indicate that the asymmetric liposomes possess a well-defined bilayer, with bilayer deformability varying depending on the components used; for instance, liposomes containing flexible components exhibit a more deformable bilayer than those made solely of lipids. The average size of the liposomes was below 200 nm, and the %EE ranged from 75% to 90%. The liposomes containing Span 80 surfactant exhibited the highest flexibility index. This technique successfully produced asymmetric liposomes with appropriate encapsulation of the DNA plasmid without degradation during the process. Future studies are expected to evaluate the cytotoxicity, transfection, and skin permeation.

Inactivation of p53 tumor suppressor functions, often through missense mutations, is essential for carcinogenesis. A sub-class of such p53 missense mutations gains new functions, including drug resistance and enhanced proliferation, in addition to its loss of function. Among the most frequent gain-of-function p53 mutants, R273H occurs in tumors of many tissue origins and imparts aggressive character and resistance to drugs to the tumor. Tumors bearing p53R273H are generally resistant to all available therapies, and need for novel interventions are urgently needed. Interaction of p53R273H with Positive Coactivator 4 (PC4), an abundant chromatin-associated protein, is essential for acquiring the gain-of-function properties. Previously, we developed a chemically modified peptide, NLS-p53(380-386), targeting PC4 that abrogated the interaction of p53R273H with PC4 and reversed many of its gain-of-function properties. We earlier demonstrated that cationic phosphatidylcholine-stearylamine (PC-SA) liposomes possess inherent anti-tumor properties. To improve efficacy, pharmacokinetics, and delivery, we entrapped the PC4-targeted peptide into PC-SA liposome. We synthesized the NLS-p53(380-386) peptide and entrapped in PC-SA liposome. We used MTT assay, confocal microscopy, flow cytometry, qRT-PCR, and western blotting to investigate the biological effects of the p53-entrapped PC-SA. Pretreatment with the PC-SA liposome entrapped peptide enhanced the chemosensitivity of widely used anticancer drug doxorubicin in cell lines bearing p53R273H mutation. The doxorubicin-induced cell-killing effect was much more enhanced when pretreated with the liposome-entrapped peptide than when pretreated with either the free peptide or the liposome alone. The liposome-encapsulated peptide is a promising formulation for developing therapies targeting tumors bearing the p53R273H.

To enhance the anticancer effects of the purslane extract, we developed a phytosomal nanocarrier with mitochondrial targeting capabilities. Initially, a phytosomal carrier was prepared and subsequently functionalized with a Szeto-Schiller (SS) peptide as, a mitochondrial-penetrating peptide, via a DSPE-PEG (2000)-malamide crosslinker. High-performance liquid chromatography analysis was conducted to quantify the amounts of quercetin and apigenin in the hydroalcoholic extract and the fractionated isolates obtained from diethyl ether, chloroform, ethyl acetate, butanol, and water. The results indicated that both polyphenols were present in the hydroalcoholic extract, with apigenin being more abundant in the ethyl acetate fraction. Dynamic light scattering measurements revealed an average particle size of 112.2 ± 6.758 nm, a narrow polydispersity index of 0.26 ± 0.005, and a zeta potential of -30.67 ± 2.894 mV. Scanning electron microscopy confirmed that the phytosomal carrier exhibited a spherical and intact morphology. The encapsulation efficiency and loading capacity of the phytosomes were found to be 97% and 12.19%, respectively, for the ethyl acetate isolate. The in vitro drug release profile demonstrated a biphasic pattern, characterized by an initial burst release followed by prolonged sustained release. Cytotoxicity assays conducted on A549 and HFF cell lines revealed that the mitochondria-targeted phytosomal nanocarrier exhibited significant apoptotic effects (69.6%) on A549 cells, showing no significant toxicity toward HFF cells when compared with phosphatidylcholine, conventional phytosomal nanocarriers, and ethyl acetate-targeted phytosomal nanocarriers. The findings from this study represent a crucial advancement in the standardization of plant extracts and the development of herbal nanomedicine using targeted phytosomes.

Cell-penetrating peptides (CPPs) with a cationic-hydrophobic character are recognized as carriers for delivering various therapeutics and diagnostic agents across cell membranes and into the cells. Among the most studied CPPs, nona-arginine (R9) exhibits superior penetration compared to its equally charged counterpart, nona-lysine (K9). This indicates that penetration ability relies not only on charge but also on the structure, distribution, and concentration of peptides, as well as the composition of lipid membranes. However, interactions of heptapeptides composed of arginine (R), lysine (K), and phenylalanine (F) residues with membranes remain poorly explored. This study sheds light on the interaction of R5F2/K5F2 on lipid membranes containing a zwitterionic lipid (phosphatidylcholine; PC) and an anionic lipid (either phosphatidylglycerol, PG or phosphatidylserine, PS) in the 90:10 molar ratio. Using differential scanning calorimetry (DSC) and temperature-dependent UV-Vis spectroscopy, we observed peptide interaction-induced changes that stabilize a particular phase of lipid bilayers, as well as their effect on the melting of the latter in terms of cooperative unit size (CUS). The distinct interaction of R5F2/K5F2 on DPPC+DPPG and DPPC+DPPS lipid bilayers revealed that the changes in lipid packing and hydrocarbon chain conformations are peptide-specific features. The peptide-induced formation of vacancies in the non-polar bilayer part is consistent with partial membrane leakage observed in giant unilamellar vesicles. This study provides new insights into the peptide-lipid interactions underlying the functionality of CPPs.