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Transdermal drug delivery of proteins is challenging because the skin acts as a natural and protective barrier. Several techniques including using the cell-penetrating peptides have been studied to increase the penetration of therapeutic proteins into and through the skin. Cell-penetrating peptides facilitate and improve the transduction of large and hydrophilic cargo molecules through plasma membrane. We have recently reported an efficient skin delivery of elastin protein in complex with a cell-penetrating peptide called Pep-1. As the biophysical characteristics of cell-penetrating peptide/protein complexes have been linked with their biological responses, in this study, we investigated biophysical properties of Pep-1/elastin complexes (ratio 10:1) stored in three temperatures (-20 °C, 4 °C and 25 °C) by photon correlation spectroscopy, circular dichroism and isothermal denaturation. We also evaluated the ability of transduction of this complex into cells and skin tissue using both fluorescence microscopy and Kodak In-Vivo FX Pro Imaging System., (© 2013 John Wiley & Sons A/S.)

Background: Transmembrane delivery of active peptides and proteins, including skin delivery of cosmeceutical proteins such as collagen and elastin, has been a challenging issue. Amphipathic cell-penetrating peptides (CPPs) have been proposed as carrier peptides to mediate cellular uptake of proteins without covalent binding., Materials and Methods: In this study, we have used a short peptide, Pep-1, as our CPP to transport elastin into fibroblast cells. Different ratios of Pep-1/elastin complexes were produced by using a fixed amount of elastin and different molar ratio of Pep-1. The ability of transduction into cells was determined by fluorescence microscopy. The characteristics of Pep-1/elastin complexes were monitored using scanning electron microscopy and photon correlation spectroscopy., Results: No cellular toxicity was observed in cells treated with Pep-1/elastin complex. Finally, we determined a Pep-1 : elastin ratio of 10 : 1 as the most effective ratio in cellular delivery of elastin., Conclusion: Pep-1 mediated fast and effective delivery of elastin as a cosmetic protein into fibroblast cells in the treatment of skin-aging symptoms., (© 2012 The International Society of Dermatology.)