Intracellular transduction of hydrophilic macromolecules has been problematic owing to the biochemical restriction imposed by lipid bilayer of the cytoplasmic membrane. Several technologies have been developed to improve the intracellular delivery of the large molecules for therapeutic purpose, including cell penetrating peptide. Cell penetrating peptides or cell permeable peptides (CPPs) were initially discovered based on the potency of certain full-length proteins or proteins to translocate across the plasma membrane. Currently, CPPs are broadly applied for intracellular delivery of biologically functional molecules in vivo and vitro, varying from small molecules, peptides, proteins, liposomes and nucleic acids. With introducing the history and characteristics of CPPs, this review will focus on the intracellular transduction mechanism and application of CPPs.
Cell Membrane ; Cell-Penetrating Peptides ; Endocytosis ; Lipid Bilayers ; Liposomes ; Nucleic Acids ; Peptides ; Proteins

BACKGROUND: Topical irritants disrupt the cutaneous permeability barrier through the removal of stratum comeum lipids. This perturbation of barrier integrity stimulates a variety of homeostatic repair responses that ultimately result in the normalization of bamer function. Object To measure the effect of desoxymethasone ointment, vaseline and hydrobase on the barrier recovery of benzalkonium chloride (BKC) imtated skin. MATERIALS AND METHODS: The left flank skin of 2-3 monthold hairless mice was treated with BKC and then desoxymethasone ointment, vaseline and hydrobase were applied. Transepidermal water loss (TEWL) was checked after 0, 3, 6, 9, 12, 15, 18, 21 and 24 hours. Electron microscopic examination was performed after 3 and 24 hours after desoxymethasone, vaseline and hydrobase had been applied. RESULTS: The recovery of TEWL was most prominantly observed in the desoxymethasone ointment treated group followed by vaseline and hydrobase. Electron microscopic examination using ruthenium tetroxide fixation revealed that secretion and numbers of lamellar bodies and complete formatice of lipid bilayers were most prominent at desoxymethasone ointment and vaseline treated group. CONCLUSION: Desoxymethasone ointment, vaseline and hydrobase can be good agents in improving bamer recovery after exposure to irritant material.
Animals ; Benzalkonium Compounds* ; Desoximetasone* ; Irritants ; Lipid Bilayers ; Mice ; Mice, Hairless* ; Permeability ; Petrolatum* ; Ruthenium ; Skin*

BACKGROUND: Acetone disrupts the cutaneous permeability barrier through the removal of stratum corneum lipids. This pertubation of barrier integrity stimulates a variety of homeostatic repair that ultimately results in the normalization of barrier function. OBJECT: To measure the effect of steroid on the barrier recovery of acetone applied skin. MATERIAL AND METHODS: The flank skin of 8~10 week old hairless mice was treated with acetone and then topical and systemic steroids were applied. Transepidermal water loss(TEWL) was checked after 0, 3, 6, 12 and 24 hours. Electron and light microscopic examination and ion capture cytochemistry were performed after 3, 6, 12 and 24 hours after systemic and topical steroids had been applied. RESULTS: The results were as follows ; 1) During 3~6 hours after experiment, the recovery rate of TEWL was most prominent in the group of acetone applied animal than other groups. 2) After 12 hours after acetone applied, formation of new stratum corneum was found in the groups of acetone applied or acetone applied skin with topical steroid application. But loss of stratum corneum was observed in the groups of high or low dose steroid injection. 3) Ruthenium tetroxide staining of acetone alone or topical steroid treated specimens after 12 hours experiment revealed that the lipid bilayer was partly impaired and fragmented. Intercellular spaces were widening and the lipid bilayer disappeared or was damaged in the groups of high or low dose steroid injection. 4) Six hours after acetone application, pattern of calcium distribution had been partially reestabilished in the group of acetone alone or topical steroid treated animals. But calcium content was still sparse and decreased from the stratum granulosum to basale in the groups of high or low dose steroid injection. CONCLUSION: In summary the present study demonstrates that steroid treatment acutely delays recovery rate of TEWL, inhibits normalization of calcium gradient or epidermal lipid synthesis that leads to abnormalities in permeability barrier homeostasis.
Acetone ; Animals ; Calcium ; Extracellular Space ; Histocytochemistry ; Homeostasis ; Lipid Bilayers ; Mice ; Mice, Hairless* ; Permeability ; Ruthenium ; Skin ; Steroids

Animals ; Heart Ventricles ; cytology ; Lipid Bilayers ; Potassium Channels ; Sarcolemma ; physiology ; Swine

BACKGROUND: The epidermal permeability barrier necessary for terrestrial life resides in the intercellular spaces of the stratum corneum and is composed of lipids. OBJECTIVE: Since strrtum corneum lipid may be important for the permeability barrier, we studied the differences and effects of experimentally altered barrier function using acetone and tape-stripping technique. METHODS: The permeability barrier of hairless mouse was disrupted by tape-stripping and acetone treatment and the recovery rate was assessed by histochemical staining, electron microscopic examination and lipid analysis. RESULTS: Although the transepidermal water loss recovered completely by 48 hours in both of the acute models, acetone treated samples seem to have on over-all better recovery rate than tape-stripped samples. The return of barrier function to normal in both tape-stripped and acetone-treated skin was accompanied by a comparable return of normal nile red and ruthenium tetroxide staining. The amount of lipid in stratum corneum paralleled both the return of barrier function towards normal and the extent of prior damage to the barrier in acetone treated skin, yet, the lipid synthesis in tape-stripped skin showed a slower return of lipid content. CONCLUSION: The difference in the recovery rate of the two acute models may be due to the fact that acetone mainly extracts intercellular lipids, whereas, tape-stripping has a prolonged effect by removal of comeocyte in addition to the intercellular lipids. This shows the importance of comeocytes as well as the intercellular lipid bilayer in the recovery of normal barrier function.
Acetone* ; Animals ; Extracellular Space ; Lipid Bilayers ; Mice ; Mice, Hairless ; Permeability ; Ruthenium ; Skin ; Water

Self-assembled lipid films provide new insights into the structure-function relationships of biomolecules at the molecular level. It has potential applications in biology and bionics. In this paper, with regard to atomic force microscopy (AFM) characterization, the surface structures and growth kinetics of self-assembled lipid films as well as their applications in high-resolution AFM imaging of surface-immobilized biomolecules such as proteins, DNA and enzymes are reviewed.
DNA ; chemistry ; Humans ; Lipid Bilayers ; chemistry ; Microscopy, Atomic Force ; methods ; Phospholipids ; chemistry ; Proteins ; chemistry

This study examined the mechanism of action of a local anesthetic, lidocaine.HCl. Energy transfer between the surface fluorescent probe, 1-anilinonaphthalene-8-sulfonic acid, and the hydrophobic fluorescent probe, 1,3-di(1-pyrenyl) propane, was used to determine the effect of lidocaine.HCl on the thickness (D) of the synaptosomal plasma membrane vesicles (SPMV) isolated from the bovine cerebral cortex, and liposomes of the total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. The thickness (D) of the intact SPMV, SPMVTL and SPMVPL were 1.044+/-0.008, 0.914+/-0.005 and 0.890+/-0.003 (arbitrary units, n=5) at 37degrees C (pH 7.4), respectively. Lidocaine.HCl decreased the thickness of the neuronal and model membrane lipid bilayers in a dose-dependent manner with a significant decrease in the thickness, even at 0.1 mM. The decreasing effect of lidocaine.HCl on the membrane thickness might be responsible for some, but not all of its anesthetic action.
Anilino Naphthalenesulfonates ; Cell Membrane ; Cerebral Cortex ; Energy Transfer ; Lipid Bilayers ; Liposomes ; Membranes ; Neurons ; Phospholipids ; Propane

Using the planar lipid bilayer method, we investigated the effect of d-tubocurarine (dTC) on the extracellular side of large-conductance Ca2+-activated K+ channel from rat brain. When the initial open probability (Po) of the channel was relatively high, dTC decreased channel activity in a concentration dependent manner. In contrast, when the initial Po was lower, sub-micro molar dTC increased channel activity by destabilizing the closed states of the channel. Further addition of dTC up to micro molar range decreased channel activity. This dual effect of dTC implicates that there exist at least two different binding sites for dTC.
Animals ; Binding Sites ; Brain* ; Lipid Bilayers* ; Molar ; Potassium Channels, Calcium-Activated* ; Rats* ; Tubocurarine*

BACKGROUND AND OBJECTIVES: Poor homing efficiency is one of the major limitations of current stem cell therapy. Magnetic bionanoparticles (MPs) obtained from Magnetospirillum sp. AMB-1 have a lipid bilayer membrane and ferromagnetic properties. We evaluated a novel priming strategy using MPs to enhance the homing of transplanted progenitor cells to target tissue. MATERIALS AND METHODS: Effects of MP on proliferation, viability, and migration of late human endothelial progenitor cells (EPCs) were examined in vitro. Additionally, effects of MP on gene and protein expression related to survival and adhesion were evaluated. Homing and angiogenic efficiency of MP transferred late EPCs was evaluated in nude mouse hindlimb ischemia model. RESULTS: Below threshold concentration, MP transfer did not influence proliferation or survival of late EPCs, but enhanced migration and trans-endothelial migration of late EPCs toward magnet. Below threshold concentration, MP transfer did not influence gene and protein expression related to survival. In the mouse hindlimb ischemia model, late EPCs treated with high dose MP (5 ug/mL) showed enhanced homing of injected late EPCs in the ischemic limb by magnet, compared to low dose MP (1 ug/mL) treated late EPCs. In addition, high dose MP transferred EPC showed significantly better improvement of perfusion in ischemic limb compared to untreated EPC. CONCLUSION: MP transfer with magnet application can be a promising novel strategy to enhance homing efficacy and outcomes of current stem cell therapy.
Animals ; Extremities ; Hindlimb ; Humans ; Ischemia ; Lipid Bilayers ; Magnetics ; Magnetospirillum ; Magnets ; Membranes ; Mice ; Mice, Nude ; Nanoparticles ; Perfusion ; Phosphorylcholine ; Stem Cells ; Transplants

BACKGROUND: The lipids of the stratum corneum, which originate from polar lipid precursors provided by the cells of the stratum granulosum via the exocytosis of lamellar bodies, with cornified cell envelope form competent epidermal barrier structurally and functionally. The ontogeny of the epidermal barrier is not clearly defined because of difficulty of sampling and methodology which defines epidermal lipids. OBJECT: From ultrastructural observation of skin samples obtained from human fetuses and newborn on serial developmental timings, we tried to clarify the sequential development of epidermal barrier. METHODS: Skin samples were obtained from 13 human fetuses from EGA(estimated gestational age) 10 to 23wks and 2 newborns. Specimens were observed by fluorescent confocal microscopy with nile red to identify the distribution of epidermal lipids, by transmission electron microscope with lanthanum to investigate the functional permeability barrier, with RuO4 to observe the intercellular lipid bilayer and morphology of lamellar bodies, with ion capture cytochemistry to investigate the formation of epidermal calcium gradient. RESULT: In nile red stain, the amount of epidermal lipid increased during fetal period. At EGA 23wks, the lipid distribution revealed linear and continuous pattern. In lanthanum tracer study, the electron dense tracer permeated all the intercellular space of the epidermis up to periderm and subepidermal space until EGA 21wks. At EGA 23wks, the tracer permeated intercellular space of epidermis weakly. It might be predicted that incomplete epidermal barrier is present at this time. In RuO4 stain, precursor of lamellar body was observed at EGA 15wks, and intercellular lipid bilayer was observed at EGA 16wks. As gestation increases, there was a steady increase in epidermal lipid bilayers. In ion capture cytochemistry, epidermal calcium gradient was first observed in follicular epidermis at EGA 20wks, and in interfollicular epidermis at EGA 23wks. From these results, it is concluded that the basic structures of epidermal barrier are formed at EGA 23wks, but it is not complete, and epidermal barrier arises first from follicular epidermis.
Calcium ; Epidermis ; Exocytosis ; Extracellular Space ; Fetus* ; Histocytochemistry ; Humans* ; Infant, Newborn* ; Lanthanum ; Lipid Bilayers ; Microscopy, Confocal ; Permeability ; Pregnancy ; Skin