Micelle-to-vesicle Transition Research Articles

Preparation of Small Unilamellar Vesicle Liposomes Using Detergent Dialysis Method.

Small unilamellar liposomes are commonly used as model biomembranes and carriers for drug and gene delivery. Although methods which employ mechanical forces or organic solvents can be used to reduce the liposome size and the number of lipid bilayers, they are not suitable options when the purpose is to incorporate biologically active proteins into lipid bilayers or to encapsulate nucleic acid into liposomes. Detergent dialysis is a simple and inexpensive procedure to produce homogeneous small unilamellar vesicles (SUVs). Lipids are solubilized by detergent at a concentration much higher than its critical micellar concentration (CMC) in an aqueous dispersion medium. Free monomer detergent molecules in the dispersion medium are removed during dialysis, while the supramolecular detergent-lipid mixed micelles (MMs) are kept inside the dialysis tubing, leading to dissociation of detergent molecules from MMs. SUVs are formed after the micelle to vesicle transition (MVT).

Macroscalar Helices Co-Assembled from Chirality-Transferring Temperature-Responsive Carbohydrate-Based Bolaamphiphiles and 1,4-Benzenediboronic Acid.

We present the first example of macroscalar helices co‐assembled from temperature‐responsive carbohydrate‐based bolaamphiphiles (CHO‐Bolas) and 1,4‐benzenediboronic acid (BDBA). The CHO‐Bolas contained hydrophilic glucose or mannose moieties and a hydrophobic coumarin dimer. They showed temperature‐responsive reversible micelle‐to‐vesicle transition (MVT) in aqueous solutions. After the binding of carbohydrate moieties with boronic acids of BDBA in their alkaline solutions, right‐handed helices were formed via the temperature‐driven chirality transfer of d‐glucose or d‐mannose from the molecular to supramolecular level. These helices were co‐assembled by unreacted BDBA, boronate esters (B−O−C bonds) between CHO‐Bolas and BDBA, as well as boroxine anhydrides (B−O−B bonds) of self‐condensed BDBA. After heating at 300 °C under nitrogen, the helices displayed excellent morphological stability. Moreover, they emitted bright blue luminescence caused by strong self‐condensation of BDBA and decomposition of coumarin dimers.

Preparation of drug-loaded small unilamellar liposomes and evaluation of their potential for the treatment of chronic respiratory diseases

The aim of the present investigation was to evaluate the influence of liposome formulation on the ability of vesicles to penetrate a pathological mucus model obtained from COPD affected patients in order to assess the potential of such vesicles for the treatment of chronic respiratory diseases by inhalation. Therefore, Small Unilamellar Liposomes (PLAIN-LIPOSOMEs), Pluronic® F127-surface modified liposomes (PF-LIPOSOMEs) and PEG 2000PE-surface modified liposomes (PEG-LIPOSOMEs) were prepared using the micelle-to-vesicle transition (MVT) method and beclomethasone dipropionate (BDP) as model drug. The obtained liposomes showed diameters in the range of 40–65 nm, PDI values between 0.25 and 0.30 and surface electric charge essentially close to zero. The encapsulation efficiency was found to be dependent on the BDP/lipid ratio used and, furthermore, BDP-loaded liposomes were stable in size both at 37 °C and at 4 °C. All liposomes were not cytotoxic on H441 cell line as assessed by the MTT assay. The liposome uptake was evaluated through a cytofluorimetric assay that showed a non-significant reduction in the internalization of PEG-LIPOSOMEs as compared with PLAIN-LIPOSOMEs. The penetration studies of mucus from COPD patients showed that the PEG-LIPOSOMEs were the most mucus-penetrating vesicles after 27 h. In addition, PEG- and PF-LIPOSOMEs did not cause any effect on bronchoalveolar lavage fluid proteins after aerosol administration in the mouse. The results highlight that PEG-LIPOSOMEs show the most interesting features in terms of penetration through the pathologic sputum, uptake by airway epithelial cells and safety profile.

Self assembly in an aqueous gemini surfactant containing sugar based (isosorbide) spacer

Effect of various stimuli (concentration, pH, temperature and salt) have been investigated, in aqueous isosorbide (sugar based) spacer based cationic gemini surfactant with hexadecyl chain (16-Isb-16), using spectroscopic and physico-chemical techniques (dynamic light scattering, DLS; nuclear magnetic resonance, NMR; transmission electron microscopy, TEM; polarizing optical microscopy; POM; zeta (ζ)-potential; count rate; pH and conductometry). An interesting phenomenon of micelle to vesicle transition (MVT) has been observed upon dilution. pH sensitive systems, including micelles and vesicles, are found in the solution without changing the pH externally. For external pH change (in the acidic range), DLS data revealed that vesicles are present near the neutral pH. The morphological transitions were confirmed by TEM and POM. ζ data show the conversion of positively charged aggregates to neutral ones. MVT has also been checked by increasing the temperature or adding a salt (NaCl, NaBr, NaNO3 or sodium salicylate; NaSal). Heating induces a transition from micelles to vesicles near ∼40–45 °C while salt addition causes a reverse effect (vesicle to micelle). The transition has been found to be dependent on concentration/nature of salt. The study provides a simple and effective way of tuning the micellar morphology.

Luminescent CdSe@ZnS nanocrystals embedded in liposomes: a cytotoxicity study in HeLa cells.

The use of fluorescent nanocrystals (NCs) as probes for bioimaging applications has emerged as an advantageous alternative to conventional organic fluorescent dyes. Therefore their toxicological evaluation and intracellular delivery are currently a primary field of research. In this work, hydrophobic and highly fluorescent CdSe@ZnS NCs were encapsulated into the lipid bilayer of liposomes by the micelle-to-vesicle transition (MVT) method. The obtained aqueous NC-liposome suspensions preserved the spectroscopic characteristics of the native NCs. A systematic study of the in vitro toxicological effect on HeLa cells of these red emitting NC-liposomes was then carried out and compared to that of empty liposomes. By using liposomes of different phospholipid composition, we evaluated the effect of the lipid carrier on the cytotoxicity towards HeLa cells. Surprisingly, a cell proliferation and death study along with the MTT test on HeLa cells treated with NC-liposomes have shown that the toxic effects of NCs, at concentrations up to 20 nM, are negligible compared to those of the lipid carrier, especially when this is constituted by the cationic phospholipid DOTAP. In particular, obtained data suggest that DOTAP has a dose- and time-dependent toxic effect on HeLa cells. In contrast, the addition of PEG to the liposomes does not alter significantly the viability of the cells. In addition, the ability of NC-liposomes to penetrate the HeLa cells was assessed by fluorescence and confocal microscopy investigation. Captured images show that NC-liposomes are internalized into cells through the endocytic pathway, enter early endosomes and reach lysosomes in 1 h. Interestingly, red emitting NCs co-localized with endosomes and were positioned at the limiting membrane of the organelles. The overall results suggest that the fluorescent system as a whole, NCs and their carrier, should be considered for the development of fully safe biological applications of CdSe@ZnS NCs, and provide essential indications to define the optimal experimental conditions to use the proposed system as an optical probe for future in vivo experiments.

The effect of temperature, composition and alcohols on the microstructures of catanionic mixtures of sodium dodecylsulfate and cetyltrimethylammonium bromide in water.

The influence of mixing protocol, composition, temperature, ageing and added alcohols on the characteristics of the microstructures of sodium dodecylsulfate (SDS) + cetyltrimethylammonium bromide (CTAB) mixtures has been investigated in this paper. In this catanionic mixture (1 weight% total surfactant content) temperature induced microstructural transition occurs, which is (i) a micelle-to-vesicle transition (MVT) if αSDS (mole fraction of SDS) = 0.7, 0.8 or 0.9 and (ii) a vesicle-to-micelle transition (VMT) if αSDS = 0.1, 0.2 or 0.3. In the mixture of αSDS = 0.7, specific conductivity and dynamic light scattering measurements also support the occurrence of MVT. Transition electron microscopy and small angle neutron scattering measurements were also made to assess the characteristics of the microstructures. Alcohols added to the mixture of αSDS = 0.7 reduced the size of the vesicle, while only monohydric alcohols suppressed the temperature induced transition indicating that the number and location of -OH groups of the alcohols have a dramatic modulating influence on the structural transition occurring in catanionic mixtures. The influence of the alcohols is explained in terms of changes produced in the dielectric constant and hydrophobicity of the medium.

Hybrid Assemblies of Fluorescent Nanocrystals and Membrane Proteins in Liposomes

Because of the growing potential of nanoparticles in biological and medical applications, tuning and directing their properties toward a high compatibility with the aqueous biological milieu is of remarkable relevance. Moreover, the capability to combine nanocrystals (NCs) with biomolecules, such as proteins, offers great opportunities to design hybrid systems for both nanobiotechnology and biomedical technology. Here we report on the application of the micelle-to-vesicle transition (MVT) method for incorporation of hydrophobic, red-emitting CdSe@ZnS NCs into the bilayer of liposomes. This method enabled the construction of a novel hybrid proteo-NC-liposome containing, as model membrane protein, the photosynthetic reaction center (RC) of Rhodobacter sphaeroides. Electron microscopy confirmed the insertion of NCs within the lipid bilayer without significantly altering the structure of the unilamellar vesicles. The resulting aqueous NC-liposome suspensions showed low turbidity and kept unaltered the wavelengths of absorbance and emission peaks of the native NCs. A relative NC fluorescence quantum yield up to 8% was preserved after their incorporation in liposomes. Interestingly, in proteo-NC-liposomes, RC is not denatured by Cd-based NCs, retaining its structural and functional integrity as shown by absorption spectra and flash-induced charge recombination kinetics. The outlined strategy can be extended in principle to any suitably sized hydrophobic NC with similar surface chemistry and to any integral protein complex. Furthermore, the proposed approach could be used in nanomedicine for the realization of theranostic systems and provides new, interesting perspectives for understanding the interactions between integral membrane proteins and nanoparticles, i.e., in nanotoxicology studies.

CO2-induced micelle to vesicle transition in zwitterionic–anionic surfactant systems

Study of micelle to vesicle transition (MVT) is of great importance from both theoretical and practical points of view. In this work, we studied the effect of compressed CO2 on the aggregation behavior of zwitterionic-anionic (DSB (dodecyl sulfonatebetaine)-AOT(sodium bis(2-ethylhexyl) sulfosuccinate)) mixed surfactants in aqueous solution by means of direct observation, turbidity, steady-state fluorescence, fluorescence quantum yield, and entrapment quantity of vesicles. Interestingly, all the methods show that compressed CO2 can induce MVT in this zwitterionic-anionic surfactant system. The CO2-induced MVT is reversible and the degree of MVT can be easily tuned by controlling the operation pressure. Further studies show that the pH decrease and dissolution of gas molecules in the surfactant film co-contribute to the MVT, and a possible mechanism for the CO2-induced MVT was proposed based on the experimental results.

Reversible Switching of a Micelle-to-Vesicle Transition by Compressed CO2

The study of the micelle-to-vesicle transition (MVT) is of great importance from both theoretical and practical points of view. Herein, we studied the effect of compressed CO(2) on the aggregation behavior of dodecyltrimethylammonium bromide (DTAB)/sodium dodecyl sulfate (SDS) mixed surfactants in aqueous solution by means of direct observation, turbidity and conductivity measurements, steady-state fluorescence, time-resolved fluorescence quenching (TRFQ), fluorescence quantum yield, and template methods. Interestingly, all these approaches showed that compressed CO(2) could induce the MVT in the surfactant system, and the vesicles returned to the micelles simply by depressurization; that is, CO(2) can be used to switch the MVT reversibly by controlling pressure. Some other gases, such as methane, ethylene, and ethane, could also induce the MVT of the surfactant solution. A possible mechanism is proposed on the basis of the packing-parameter theory and thermodynamic principles. It is shown that the mechanism of the MVT induced by a nonpolar gas is different from the MVT induced by polar and electrolyte additives.

Micelle‐to‐Vesicle Transition Induced by Organic Additives in Catanionic Surfactant Systems

A micelle-to-vesicle transition (MVT) induced by the addition of a series of apolar hydrocarbons (n-butylbenzene, n-hexane, n-octane, and n-dodecane) to the catanionic surfactant system n-dodecyltriethylammonium bromide/sodium n-dodecylsulfate (DTEAB/SDS) has been investigated for the first time by means of rheology and turbidity measurements, dynamic light scattering (DLS), and transmission electron microscopy (TEM). Interestingly, a MVT can take place within certain micellar regions, which are dependent on the structure and chain length of the hydrocarbon. However, these hydrocarbons are unable to induce a MVT in another catanionic surfactant system, namely, n-dodecyltriethylammonium bromide/sodium n-dodecylsulfonate (DTEAB/SDSO(3)), in which the molecular interactions are weaker than in the DTEAB/SDS system. On the other hand, polar additives, such as n-octanol and n-octylamine, exhibit much higher efficiency and activity in inducing MVT than hydrocarbons in both DETAB/SDS and DTEAB/SDSO(3). Moreover, DLS, TEM, and time-resolved fluorescence quenching (TRFQ) results demonstrate that the ratio of vesicles to micelles in the system can be actively controlled by addition of polar additives. Possible mechanisms for the above phenomena are presented, and the potential application of controllable micelle/vesicle systems in the synthesis of tailored bimodal mesoporous materials is discussed.

Micelle-to-Vesicle Transition of an Iron-Chelating Microbial Surfactant, Marinobactin E

Small-angle neutron scattering (SANS) and dynamic light scattering (DLS) techniques have been applied to study the self-assembly processes of a microbially produced siderophore, marinobactin E (ME). ME is one of a series of marinobactins A-E that facilitate Fe(III) acquisition by the source bacterium through coordination of Fe(III) by the marinobactin headgroup. ME is a six-amino-acid peptide amphiphile appended by palmitic acid (C16), and differs only in the nature of the fatty acid moiety from the other marinobactins. Apo-ME (uncoordinated ME) assembles to form micelles with an average diameter of 4.0 nm. Upon coordination of one equivalent of Fe(III), the mean micellar diameter of Fe(III)-ME shrinks to approximately 2.8 nm. However, in the presence of excess Fe(III), Fe(III)-ME undergoes a micelle-to-vesicle transition (MVT). At a small excess of Fe(III) over Fe(III)-ME (i.e., <1.2 Fe(III)/ME), a fraction of the Fe(III)-ME micelles rearrange into approximately 200 nm diameter unilamellar vesicles. At even greater Fe(III)/ME ratios (e.g., 2-3) multilamellar aggregates begin to emerge, consistent with either multilamellar vesicles or lamellar stacks. The MVT exhibited by ME may represent a unique mechanism by which marine bacteria may detect and sequester iron required for growth.

Heating-Induced Micelle to Vesicle Transition in the Cationic−Anionic Surfactant Systems: Comprehensive Study and Understanding

Heating-induced micelle to vesicle transition (MVT), which has been rarely reported in surfactant systems, was systemically studied in a number of mixed cationic-anionic surfactant systems. According to the turbidity measurements, the investigated systems can be divided into two classes: Class A and B. Heating-induced MVT was observed in Class A at certain total surfactant concentrations and mixed surfactant ratios, while no such transition was found in Class B. Further investigations revealed that the heating-induced MVT is more likely to take place in the cationic-anionic surfactant systems with relatively stronger molecule interaction and larger micelle aggregation number. The effects of several physicochemical factors, such as the variation of mixed surfactant ratios and the addition of n-decanol on the heating-induced MVT, were also studied.