Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane

Cell-penetrating peptide (CPP) can directly penetrate the cytosol (cytolysis) and is expected to be a potent vector for a drug delivery system (DDS). Although there is general agreement that CPP cytolysis is related to dynamic membrane deformation, a distinctive process has yet to be established. Here, we report the key process and factors controlling CPP cytolysis. To elucidate the task, we have introduced trypsin digestion of adsorbed CPP onto giant unilamellar vesicle (GUV) to quantify the adsorption and internalization (cytolysis) separately. Also, the time-course analysis was introduced for the geometric calculation of adsorption and internalization amount per lipid molecule consisting of GUV. As a result, we found that adsorption and internalization assumed to occur successively by CPP molecule come into contact with membrane lipid. Adsorption is quick to saturate within 10 min, while cytolysis of each CPP on the membrane follows successively. After adsorption is saturated, cytolysis proceeds further linearly by time with a different rate constant that is dependent on the osmotic pressure. We also found that temperature and lipid composition influence cytolysis by modulating lipid mobility. The electrolyte in the outer media is also affected as a chemical mediator to control CPP cytolysis by following the Hoffmeister effect for membrane hydration. These results confirmed the mechanism of cytolysis as temporal and local phase transfer of membrane lipid from Lα to Mesh1, which has punctured bilayer morphologies.

Influence of Carbon Nanosheets on the Behavior of 1,2-Dipalmitoyl-sn-glycerol-3-phosphocholine Langmuir Monolayers

Carbon nanomaterials are widespread in the atmospheric aerosol as a result of the combustion processes and their extensive industrial use. This has raised many question about the potential toxicity associated with the inhalation of such nanoparticles, and its incorporation into the lung surfactant layer. In order to shed light on the main physical bases underlying the incorporation of carbon nanomaterials into lung surfactant layers, this work has studied the interaction at the water/vapor interface of carbon nanosheets (CN) with Langmuir monolayers of 1,2-Dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), with this lipid being the main component of lung surfactant layers and responsible of some of the most relevant features of such film. The incorporation of CN into DPPC Langmuir monolayers modifies the lateral organization of the DPPC at the interface, which is explained on the basis of two different effects: (i) particles occupy part of the interfacial area, and (ii) impoverishment of the lipid composition of the interface due to lipid adsorption onto the CN surface. This results in a worsening of the mechanical performance of the monolayers which may present a negative impact in the physiological performance of lung surfactant. It would be expected that the results obtained here can be useful as a step toward the understanding of the most fundamental physico-chemical bases associated with the effect of inhaled particles in the respiratory cycle.

Mass spectrometric imaging of in vivo protein and lipid adsorption on biodegradable vascular replacement systems

Cardiovascular diseases present amongst the highest mortality risks in Western civilization and are frequently caused by arteriosclerotic vessel failure.