Search Results (1,220)

Transferosomes are liposome-derived ultradeformable vesicles designed to improve drug delivery across restrictive biological barriers, particularly in non-invasive administration routes. Their structure is based on phospholipid bilayers modified with edge activators, usually surfactants or bile salts, which increase membrane flexibility while preserving vesicular organization. This balance between deformability and stability distinguishes transferosomes from conventional liposomes and has supported their use in dermal, transdermal, ocular, nasal, buccal, and other mucosal delivery systems. However, despite extensive experimental interest, the field remains limited by inconsistent terminology, heterogeneous formulation strategies, non-harmonized deformability assays, and incomplete translation from laboratory formulations to clinically relevant products. This review critically examines transferosomes from a formulation-development perspective, focusing on the relationship between lipid composition, edge-activator selection, vesicle properties, deformability, drug release, and biological performance. Particular attention is given to critical quality attributes, analytical characterization, mechanistic interpretations of barrier interaction, and the unresolved debate between intact vesicle penetration, drug-release-dominated delivery, and barrier perturbation. Transferosomes are also positioned in comparison with conventional liposomes, ethosomes, and transethosomes. Finally, the review identifies key unmet needs related to standardization, reproducibility, scalability, storage stability, and regulatory uncertainty. By integrating formulation design with mechanistic and translational analysis, this review aims to clarify when transferosomes offer a genuine delivery advantage and which parameters must be controlled to support their further pharmaceutical development. Full article

Plant-derived exosome-like nanoparticles (PELNs), a subset of extracellular vesicle (EV) secreted by plant cells, have emerged as revolutionary biomaterial with broad applications in biomedicine, agriculture, and nanotechnology. Structurally, PELNs feature a phospholipid bilayer homologous to plant cell membranes, encapsulating bioactive components such as proteins, nucleic acids, lipids, and secondary metabolites. The native structure of PELNs endows them with enhanced bioavailability, reduced immunogenicity, and improved barrier penetration for precise tissue delivery. Recent studies highlight the cross-kingdom therapeutic potential of PELNs in mammals, including antitumor, anti-inflammatory, tissue repair, immunomodulation and so on. This review comprehensively summarized recent advancements in PELN research, including innovative isolation techniques, molecular characterization, their roles in drug delivery and disease therapy. We also discussed challenges in standardization, scalability, and regulatory frameworks which could provide future perspectives for translating PELNs into clinical and industrial applications. Full article

Bacterial extracellular vesicles (BEVs) are nanoscale spherical lipid bilayer structures secreted by bacteria, including outer membrane vesicles (OMVs) released by Gram-negative bacteria and membrane vesicles (MVs) produced by Gram-positive bacteria. Although the biogenesis of BEVs requires substantial energy expenditure, these vesicles provide bacteria with strategic advantages in the evolutionary interplay between pathogens and hosts. BEVs contribute to bacterial adaptation to environmental stress by remodeling membrane components, eliminating toxic substances, promoting biofilm formation, and mediating the interbacterial transfer of antibiotic resistance determinants. They can also function as decoys to protect bacteria from bacteriophage or antibiotic attack, deliver virulence factors, modulate host immune responses to facilitate bacterial colonization, and mediate interspecies competition. This review summarizes the central roles of BEVs as bacterial mediators of environmental responses, with particular emphasis on their involvement in immune regulation, environmental adaptation, and interspecies competition, thereby providing new insights into pathogen evolutionary strategies. Full article

HP1090 is a short, cationic, amphipathic peptide derived from scorpion venom and previously described as a membrane-active antiviral compound. Here, we primarily characterize the antiviral activity of HP1090 and assess whether additional antibacterial effects are consistent with membrane-disruptive properties. Chemically synthesized HP1090 exhibited dose-dependent virucidal activity against multiple enveloped viruses, including herpes simplex virus type 1 and 2 (HSV-1, HSV-2), human immunodeficiency virus type 1 (HIV-1), and Zika virus (ZIKV), with IC₅₀ values ranging from 14.7 to 56.1 µg/mL. No activity was observed against the non-enveloped human rhinovirus 14 (HRV14), suggesting strict dependence on a viral lipid envelope. Consistent with a membrane-targeting mechanism, HP1090 induced rapid and concentration-dependent permeabilization of virus-like liposomes. HP1090 also displayed antibacterial activity against selected clinically relevant pathogens in agar-based growth inhibition assays. However, antibacterial effects required substantially higher concentrations (>125 µg/mL) and varied between bacterial species, with some strains showing little or no susceptibility. Membrane permeabilization assays in Listeria monocytogenes demonstrated disruption of bacterial membrane integrity as a contributing mechanism. No cytotoxicity was observed on mammalian cell lines at effective antiviral concentrations. Together, these findings establish HP1090 as a membrane-active venom peptide and, by linking envelope-dependent viral inactivation with bacterial membrane permeabilization, support a shared biophysical mode of action relevant to the development of membrane-targeting anti-infectives. Full article

Bacterial outer membrane vesicles (OMVs) are spherical structures made up of a double layer, they are each nanostructured (20–300 nm), and they are released from all populations of Gram-negative bacteria. The purpose of this review is to synthesize a comprehensive summary of the current state of knowledge about OMV biogenesis, function in biology, and application to biomedical engineering. Using these three known biogenesis mechanisms as a basis for this review, we discuss the mechanisms of OMV biogenesis that have been described as conserved: (1) disruption of outer membrane–peptidoglycan links. (2) periplasmic stress-driven adaptive release is associated with bilayer lipid asymmetry and the use of signaling molecules. OMVs are considered to be “public goods” for the microbe, allowing for nutrient acquisition, resistance to antibiotics, and the potential for horizontal gene transfer between microbes. OMVs exhibit a different duality at the interface of the pathogen host, where the pathogenic OMV is the delivery vehicle for virulence factors and pathogen-associated molecular patterns (PAMPs) leading to host immune response, while the symbiotic OMV (e.g., those produced by Bacteroides fragilis (Bact. fragilis)) promote regulatory T cell differentiation and mucosal tolerance. The review also addresses the various techniques currently available to isolate OMVs (e.g., ultracentrifugation and size-exclusion chromatographic techniques) and presents engineered/alloying strategies (e.g., genetic modifications to tolR/msbB and surface functionalization) to enhance the viability, safety, and specificity of OMVs for biomedical delivery. Finally, the review addresses significant obstacles related to standardization, batch variation, and in vivo safety associated with synthetic or personalized therapeutics based on OMVs, providing some recommendations for future research in this area. Full article

The most recent research has demonstrated that oscillatory nano-structures found on the lumenal walls of deep cerebral veins likely contribute significantly to the regulation of the function of deep cerebral veins. The oscillatory nano-structures consist of very small, intricately organized “nanoflaps,” each consisting of a hinge element with an attached lipid bilayer architecture. These nanoflaps have distinct mechanical properties, are in close proximity to mechanically sensitive protein assemblies, and therefore it is hypothesized that the nanoflaps generate rhythmic oscillations that control the distribution of both pressure and fluid flow through the veins and also regulate the metabolic condition of the surrounding tissue. In addition, the behavior of the nanoflaps indicate that there exists a hitherto unappreciated level of venous biomechanics at the nanometer scale that regulates the hydraulic stability of the veins and may also contribute to the structural integrity of the surrounding tissues. The purpose of this review is to provide a theoretical framework for understanding the recent discoveries of the structure, oscillation and hydrodynamic effects of nanoflaps, including resonance drift, waveform irregularity, and multi-scale biomechanical interactions. Additionally, this review will present the idea that disruption of the normal oscillatory processes that occur in the nanoflaps may lead to the development of abnormal micro-environments in the early stages of neurodegenerative diseases, abnormalities of compliance, dysautonomic states, traumatic injury and micro-circulatory stress. Finally, this review will describe several pharmacological strategies that may be used to stabilize the oscillations generated by the nanometer-scale oscillatory nano-structure by modifying the torque applied to the hinge, the viscoelasticity of the membrane and the feedback pathways for mechanotransduction. Full article

Background: Cytochrome P450 3A4 (CYP3A4) is a key membrane-anchored drug-metabolizing enzyme. Its expression and purification in heterologous systems are severely hindered by low yield and detergent-induced structural inactivation. Although extracellular vesicles (EVs) provide an ideal natural lipid bilayer environment to stabilize membrane proteins, targeted loading remains challenging. The ESCRT and ALIX-binding region (EABR) of CEP55 can efficiently recruit core components of the endosomal sorting complex (ESCRT) to mediate membrane fission. Objectives: This study used the EABR motif to drive the targeted vesicular secretion of CYP3A4, thereby establishing a novel membrane protein engineering platform. Methods and Results: EABR was fused with fluorescent protein, confirming its specific mediation of vesicular secretion. Recombinant plasmids of EABR/CYP3A4 and its reverse mutant (R-EABR) were transfected into HEK293T cells. Western blot and midazolam-based metabolic assays showed that forward EABR significantly enhanced CYP3A4 expression and EV secretion, while R-EABR lost exocytosis function. EVs isolated by ultracentrifugation verified EABR’s role in recruiting ESCRT and improving CYP3A4 activity. Conclusions: Forward CEP55-EABR specifically and efficiently drives vesicular encapsulation of CYP3A4, enhancing its expression and secretion. This ESCRT-mediated strategy avoids destructive purification, provides a stable lipid-rich bioreactor for CYP3A4, and has great translational potential in high-throughput in vitro drug metabolism and screening platforms. Full article

Gymnema lactiferum (G. lactiferum) is a medicinal plant that contains potent bioactive phytochemicals, which are prone to degradation during processing and digestion. In this study, G. lactiferum extract was prepared and encapsulated into soy lecithin primary liposomes (PL) and then coated with chitosan to form secondary liposomes (chitosomes, CS) to enhance stability. Physicochemical characteristics, morphology, thermal behavior, and storage stability were evaluated. Extract loading significantly (p < 0.05) increased the mean diameter of PL from 128.6 nm to 146.3 nm and of CS from 359.1 nm to 408.9 nm compared with unloaded liposomes. Both liposomal systems exhibited homogeneous size distributions and good colloidal stability, with zeta potentials of −39.4 mV for PL and +35.8 mV for CS and low polydispersity indices (<0.25) for both systems. Transmission electron microscopy demonstrated predominantly spherical morphologies in both systems. Chitosan coating significantly (p < 0.05) improved both encapsulation efficiency (77.3%) and encapsulation yield (82.4%) compared with PL (73.7% and 79.1%, respectively). HPLC-based quantification using rutin as a reference analyte further indicated EE-R% values of 59.8% for PL-GE and 70.3% for CS-GE, supporting improved extract retention following chitosan coating. Fourier transform infrared spectroscopy confirmed successful encapsulation without apparent chemical alterations or reactions. Differential scanning calorimetry indicated that chitosan coating modified the thermal transition behavior of the liposomal membrane, consistent with altered bilayer packing and increased membrane fluidity, while incorporation of the extract partially restored thermal order within the coated system. Overall, chitosan coating effectively enhanced the encapsulation efficiency, stability, and yield of G. lactiferum extract-loaded liposomes towards their incorporation into functional food formulations. Full article

Extracellular vesicles (EVs) are attracting considerable interest due to their important role in cell signaling. However, their nanosized scale, complexity, and heterogeneity make even morphological characterization challenging. Only with the recent advances in cryogenic electron microscopy (cryo-EM), together with the increasing availability of cryo-electron microscopes, has it become possible to visualize the native structure of EVs. In this study we performed an in-depth cryo-EM analysis of EVs derived from four glioblastoma multiforme (GBM) cell lines (U87MG, U373MG, U251MG, and T98G), highlighting the morphological changes induced by temozolomide (TMZ) chemotherapeutic treatment. Size, shape, circularity, concentricity, membrane thickness, and electron density of EVs were analyzed. The key characteristic revealed by cryo-EM was that EVs can be enclosed not only by one membrane bilayer, but also by two or more bilayers (double-layered vesicles, DVs; and multilayered vesicles, MVs). Overall, TMZ treatment substantially modified both the morphology and production of EVs, decreasing the percentage of single-layered vesicles (SVs) while increasing that of DVs and MVs, as well as reducing the electron density of the EV cargo. Morphometric and morphological information can shed light on the contribution of EVs to tumor progression, metabolism, drug resistance, and immune evasion. Full article

The cell wall (CW) of Mucoromycota has a unique chitin/chitosan complex, unlike chitin/glucan complex in Ascomycota. Under cell wall stress (CWS), induced by azo dyes, ascomycetes increase the amount of CW chitin. This study analyzes the response of Mucor flavus to CWS, induced by Congo red and Calcofluor white. It was found that azo dyes significantly reduced the biomass yield and inhibited apical growth and branching but did not lead to an increase in the amount of CW chitin/chitosan, neutral polysacchrides and cytosol osmolytes. Non-bilayer phosphatidic acids and phosphatidylethanolamines dominated in the control membrane lipids, but the proportion of bilayer phosphatidylcholines did not exceed 5%. Under CWS, the proportion of phosphatidic acids increased, while the proportion of phosphatidylethanolamines decreased and the degree of unsaturation of phospholipids increased. Storage lipids in the control were represented by mono-, di- and triacylglycerides and free fatty acids. Under CWS, the proportion of diacylglycerides increased significantly, while the proportion of triacylglycerides decreased. Thus, the CWS response of M. flavus consisted of significant changes in growth and the composition of membrane and storage lipids, but the amount of CW chitin/chitosan and cytosol osmolytes did not increase, which is different from the response of ascomycetes. Full article

Xanthophylls are oxygenated carotenoids widely distributed in photosynthetic microorganisms, plants, algae, and certain invertebrates, where they function as key photoprotective and antioxidant pigments. Among them, xanthophylls containing vicinal 1,2-diol moieties exhibit unique chemical reactivity that enables reversible coordination with boron species naturally present in marine and terrestrial environments. The formation of cyclic borate esters between boron and diol-containing xanthophylls induces structural and electronic modifications that may enhance pigment stability and functional performance. Emerging evidence suggests that boron–xanthophyll complexes display improved resistance to photooxidative degradation, enhanced singlet oxygen quenching capacity, and increased radical-scavenging activity compared with their uncomplexed counterparts. In addition, boron coordination can influence molecular conformation, polarity, and supramolecular organization within lipid bilayers, thereby promoting membrane stabilization under conditions of high light exposure and oxidative stress. Together, these effects indicate a cooperative role of boron complexation in amplifying the intrinsic photoprotective and antioxidant properties of xanthophylls. A deeper understanding of the structural basis and biological implications of boron–xanthophyll interactions may provide new insights into adaptive stress tolerance in marine and photosynthetic organisms, as well as guide the development of advanced photoprotective systems for biomedical and technological applications. Full article

Life demonstrates remarkable homochirality of its major building blocks: nucleic acids, amino acids, sugars, and phospholipids. Phospholipid bilayer vesicles (liposomes) are formed at the water/air interface from Langmuir layers and contain ribose, a constituent of primordial water. Although the primordial ribose was initially racemic, life, as we know it, is homochiral, with d-ribose and its derivatives as the predominant forms. The phospholipid membrane’s permeability to d-ribose, together with ribose’s interaction with the bilayer’s charged phosphate groups, leads to ribose phosphorylation, yielding d-ribose-5-phosphate. Once inside, the d-ribose-5-phosphate molecules cannot cross the membrane. A similar path also exists for l-ribose, but with a lower rate. Therefore, overall, this process is enantioselective, favoring the buildup of d-ribose over l-ribose. Through liposome fusion, fission, and self-replication, this eventually leads to the Darwinian evolution of these structures and to the conversion of d-ribose-5-phosphate into complex functional molecules, such as ribozymes and RNA, and eventually into DNA, all of which inherit d-ribose’s chirality. Full article

Bipolar cancellation (BPC) is an efficiency-limiting phenomenon in bipolar nanosecond pulsed electric field (nsPEF) exposures, in which the second, opposite-polarity phase reduces or partially reverses the electroporation induced by the first phase. Nisin, a cationic antibiotic peptide, has been reported to interact with lipid membranes in bacterial systems and artificial bilayer models, where it may contribute to membrane destabilization and increased permeability during pulsed electric field exposure. This study investigated whether nisin may enhance the efficacy of bleomycin electrochemotherapy (ECT) in the presence of bipolar nanosecond pulses, which are typically associated with pronounced BPC effects. Pulsed electric field (PEF) parameters and drug concentrations were selected based on preliminary viability and Yo-Pro-1 uptake experiments in CLS-354 human squamous cell carcinoma cells. To evaluate the effect of nisin, cell viability and membrane permeabilization were assessed following exposure to 300 ns pulses across a range of bipolar PEF protocols, with or without nisin, while identical unipolar pulses were used for comparison. Nisin (50 µg/mL) increased membrane permeabilization across the tested range of field amplitudes (9–15 kV/cm) and burst repetition frequencies (0.1–1.66 MHz). The presence of nisin was also associated with increased efficacy of bleomycin-based ECT under both unipolar and symmetrical bipolar PEF conditions. Under the optimized parameters tested (13 kV/cm; 150 pulses of 300 ns at 1.66 MHz), bipolar nsPEFs in combination with nisin reached levels of efficacy comparable to those observed with unipolar waveforms, suggesting a potential attenuation of bipolar cancellation effects. Full article

Bacterial membrane vesicles (BMVs), encompassing outer membrane vesicles (OMVs) released from Gram-negative bacteria and extracellular vesicles (EVs) released from Gram-positive bacteria, have emerged as promising vaccine platforms owing to their intrinsic immunostimulatory properties and capacity to deliver a wide range of antigens. Although conventional vaccines effectively prevent infectious diseases, their long-term efficacy is often limited by antigenic variation and reliance on a restricted number of licensed adjuvants. BMVs, as self-adjuvanting systems, enable both antigen delivery and innate immune activation. BMVs are nanoscale lipid bilayer structures enriched with pathogen-associated molecular patterns (PAMPs), facilitating their recognition and uptake by antigen-presenting cells. This leads to the activation of pattern recognition receptors and the induction of pro-inflammatory cytokines, type I interferons, and adaptive immune responses, including antibody production and Th1- and Th17-biased cellular immunity. Recent studies highlight the versatility of BMVs as vaccine platforms across bacterial, fungal, and viral infection models. BMVs induce protective immunity by promoting both systemic and mucosal immune responses, thereby reducing bacterial burden and limiting pathogen colonization across diverse infection models. These properties have supported their application in viral vaccine development, including influenza and SARS-CoV-2, with the potential to enhance mucosal immunity. Despite these advantages, challenges remain in standardization, safety, and antigen-loading efficiency. Engineered BMVs incorporating protein or mRNA antigens may further enhance antigen presentation and CD8⁺ T cell responses. This review summarizes the biological features, immunological mechanisms, and future potential of BMVs in vaccine development. Full article