Phase behavior of selected artificial lipids

The flexibility of biomembranes is based on the physical-chemical properties of their main components - glycerophospholipids. The structure of these modular amphiphilic molecules can be modified through organic synthesis making it possible to study specific physical-chemical effects in detail. In particular, the roles of the hydrophobic tails of the phospholipids and their hydrophobic/hydrophilic interfacial backbone on the phase behaviour are highlighted. The spatial orientation of the glycerol backbone changes from sn-1,2 to sn-1,3 phospholipids leading to an increase of the in-plane area of the molecule. The larger distance between the hydrophobic tails can lead to membrane leaflet interdigitation. The introduction of methyl side groups in the hydrophobic tails increases the fluidity of the bilayer. Depending on the position of the methyl branches partial interdigitation is observed. In the case of bolaamphiphiles, methyl side groups have a similar effect on the fluidity, but interdigitation cannot occur

The use of shear stress for targeted drug delivery

Stenosed segments of arteries significantly alter the blood flow known from healthy vessels. In particular, the wall shear stress at critically stenosed arteries is at least an order of magnitude higher than in healthy situations. This alteration represents a change in physical force and might be used as a trigger signal for drug delivery. Mechano-sensitive drug delivery systems that preferentially release their payload under increased shear stress are discussed. Therefore, besides biological or chemical markers, physical triggers are a further principle approach for targeted drug delivery. We hypothesize that such a physical trigger is much more powerful to release drugs for vasodilation, plaque stabilization, or clot lysis at stenosed arteries than any known biological or chemical one

Artificial phospholipids and their vesicles

Phospholipids are at the heart and origin of life on this planet. The possibilities in terms of phospholipid self-assembly and biological functions seem limitless. Nonetheless, nature exploits only a small fraction of the available chemical space of phospholipids. Using chemical synthesis, artificial phospholipid structures become accessible, and the study of their biophysics may reveal unprecedented properties. In this article, the recent advances by our work group in the field of chemical lipidology are summarized. The family of diamidophospholipids is discussed in detail from monolayer characterization to the formation of faceted vesicles, culminating in the template-free self-assembly of phospholipid cubes and the possible applications of vesicle origami in modern personalized medicine

Structure and conserved function of iso-branched sphingoid bases from the nematode Caenorhabditis elegans

Sphingolipids are bio-active metabolites that show structural diversity among eukaryotes. They are essential for growth of all eukaryotic cells but when produced in an uncontrolled manner can lead to cell death and pathologies including auto-immune reactions, cancer, diabetes and neurodegeneration. Caenorhabditis elegans is an important genetic model organism both to find new drug-targets against parasitic nematodes and to study the conserved roles of sphingolipids in animals like their essential functions in very basic cellular processes ranging from maintenance of cell polarity and mitochondrial repair to growth and survival. C. elegans produces sphingoid bases which are structurally distinct from those of other animals as both iso- and anteiso-branched species have been reported. Using metabolic labeling we show that most worm sphingoid bases are iso-branched. We have synthesized the nematode- specific C17 iso-branched sphinganine and its 1-deoxy analogue and could show that both the iso-branch and the 1-hydroxyl group are essential to form functional nematode sphingolipids which are needed to maintain intestinal function. The organism specificity was examined by complementation experiments in Saccharomyces cerevisiae yeast cells lacking sphingoid base synthesis. We found that iso-branched sphingoid base did not support growth of mutant cells and was toxic to wild type yeast. 1-Deoxy sphingolipids have been linked to the hereditary disease HSAN1A and other metabolic disorders including diabetes. We found that in C. elegans the 1-deoxy analogue cannot rescue the intestinal phenotype caused by sphingoid base depletion. In fact, in wild-type animals with normal sphingoid base biosynthesis, exogenous 1- deoxy analogue had a disruptive effect on apical cytoskeletal organization of intestinal cells indicating that atypical bases can interfere with normal sphingolipid function

X-ray micro computed tomography for the visualization of an atherosclerotic human coronary artery

Atherosclerosis refers to narrowing or blocking of blood vessels that can lead to a heart attack, chest pain or stroke. Constricted segments of diseased arteries exhibit considerably increased wall shear stress, compared to the healthy ones. One of the possibilities to improve patient's treatment is the application of nano-therapeutic approaches, based on shear stress sensitive nano-containers. In order to tailor the chemical composition and subsequent physical properties of such liposomes, one has to know precisely the morphology of critically stenosed arteries at micrometre resolution. It is often obtained by means of histology, which has the drawback of offering only two-dimensional information. Additionally, it requires the artery to be decalcified before sectioning, which might lead to deformations within the tissue. Micro computed tomography (muCT) enables the three-dimensional (3D) visualization of soft and hard tissues at micrometre level. muCT allows lumen segmentation that is crucial for subsequent flow simulation analysis. In this communication, tomographic images of a human coronary artery before and after decalcification are qualitatively and quantitatively compared. We analyse the cross section of the diseased human coronary artery before and after decalcification, and calculate the lumen area of both samples

Bilayer Properties of 1,3-Diamidophospholipids

A series of 1,3-diamido phosphocholines was synthesized, and their potential to form stable bilayers was investigated. Large and giant unilamellar vesicles produced from these new lipids form a wide variety of faceted liposomes. Factors such as cooling rates and the careful choice of the liposome preparation method influence the formation of facets. Interdigitation was hypothesized as a main factor for the stabilization of facets and effectively monitored by small-angle X-ray scattering measurements

Rigid urea and self-healing thiourea ethanolamine monolayers

A series of long-tail alkyl ethanolamine analogs containing amide-, urea-, and thiourea moieties was synthesized and the behavior of the corresponding monolayers was assessed on the Langmuir–Pockels trough combined with grazing incidence X-ray diffraction experiments and complemented by computer simulations. All compounds form stable monolayers at the soft air/water interface. The phase behavior is dominated by strong intermolecular headgroup hydrogen bond networks. While the amide analog forms well-defined monolayer structures, the stronger hydrogen bonds in the urea analogs lead to the formation of small three-dimensional crystallites already during spreading due to concentration fluctuations. The hydrogen bonds in the thiourea case form a two-dimensional network, which ruptures temporarily during compression and is recovered in a self-healing process, while in the urea clusters the hydrogen bonds form a more planar framework with gliding planes keeping the structure intact during compression. Because the thiourea analogs are able to self-heal after rupture, such compounds could have interesting properties as tight, ordered, and self-healing monolayers

Against the rules: pressure induced transition from high to reduced order

Envisioning the next generation of drug delivery nanocontainers requires more in- depth information on the fundamental physical forces at play in bilayer membranes. In order to achieve this, we combine chemical synthesis with physical–chemical analytical methods and probe the relationship between a molecular structure and its biophysical properties. With the aim of increasing the number of hydrogen bond donors compared to natural phospholipids, a phospholipid compound bearing urea moieties has been synthesized. The new molecules form interdigitated bilayers in aqueous dispersions and self-assemble at soft interfaces in thin layers with distinctive structural order. At lower temperatures, endothermic and exothermic transitions are observed during compression. The LC1 phase is dominated by an intermolecular hydrogen bond network of the urea moieties leading to a very high chain tilt of 52°. During compression and at higher temperatures, presumably this hydrogen bond network is broken allowing a much lower chain tilt of 35°. The extremely different monolayer thicknesses violate the two-dimensional Clausius–Clapeyron equation

Correlation of surface pressure and hue of planarizable push-pull chromophores at the air/water interface

It is currently not possible to directly measure the lateral pressure of a biomembrane. Mechanoresponsive fluorescent probes are an elegant solution to this problem but it requires first the establishment of a direct correlation between the membrane surface pressure and the induced color change of the probe. Here, we analyze planarizable dithienothiophene push–pull probes in a monolayer at the air/water interface using fluorescence microscopy, grazing-incidence angle X-ray diffraction, and infrared reflection–absorption spectroscopy. An increase of the lateral membrane pressure leads to a well-packed layer of the ‘flipper’ mechanophores and a clear change in hue above 18 mN/m. The fluorescent probes had no influence on the measured isotherm of the natural phospholipid DPPC suggesting that the flippers probe the lateral membrane pressure without physically changing it. This makes the flipper probes a truly useful addition to the membrane probe toolbox