ABSTRACT
Short-interfering RNAs (siRNAs) are a potential strategy for the treatment of cutaneous diseases. In this context, liquid crystalline nanoparticles functionalized with specific proteins and peptide-transduction domains (PTDs), which act as penetration enhancers, are a promising carrier for siRNA delivery through the skin. Herein, hexagonal phase liquid crystal nanoparticles based on monoolein (MO) and/or oleic acid (OA) containing (or lacking) the cationic polymer polyethylenimine (PEI) and the cationic lipid oleylamine (OAM) were functionalized with the membrane transduction peptides transcriptional activator (TAT) or penetratin (PNT). These nanoparticles were complexed with siRNA and characterized by particle size, polydispersity, zeta potential, complexation efficiency and siRNA release. The formulations containing cationic agents presented positive zeta potentials, sizes on the nanometer scale, and complexed siRNAs at concentrations of 10 µM; these agents were successfully released in a heparin competition assay. Cell culture studies demonstrated that nanoparticles composed of MO:OA:PEI functionalized with TAT were the most efficient at transfecting L929 cells, and the uptake efficiency was enhanced by TAT peptide functionalization. Thereafter, the selected formulations were evaluated for in vivo skin irritation, penetration and in vivo efficacy using a chemically induced inflammatory animal model. These nanoparticles did not irritate the skin and provided higher siRNA penetration and delivery into the skin than control formulations. Additionally, efficacy studies in the animal model showed that the association of TAT with the nanodispersion provided higher suppression of tumor necrosis factor (TNF)-α. Thus, the development of liquid crystalline nanodispersions containing TAT may lead to improved topical siRNA delivery for the treatment of inflammatory skin diseases.
Subject(s)
Cell-Penetrating Peptides/pharmacology , Gene Silencing , Inflammation Mediators/metabolism , Liquid Crystals/chemistry , Nanoparticles/chemistry , RNA, Small Interfering/administration & dosage , Tumor Necrosis Factor-alpha/genetics , Administration, Topical , Animals , Anions , Cell Survival/drug effects , Chemistry, Pharmaceutical , Electrophoresis, Agar Gel , Gene Silencing/drug effects , Mice , Mice, Hairless , Permeability/drug effects , Skin/drug effects , Skin/pathology , Skin Diseases/pathology , Transfection , UltrasonicsABSTRACT
AIM: An intra-buccal acrylic bioadhesive device designated for drug programmed release that can stay adhered to dental enamel, and also on removable prosthetic restorations, with preventive and/or therapeutic purpose for a large clinical applications based on polymethyl methacrylate/methyl methacrylate/2 hydroxyethyl methacrylate (PMMA/MMA/HEMA) was developed, using the sodium fluoride as an active principle. This bioadhesive was evaluated for its shear bond strength when bonded with different adhesive systems. METHODS: Two substrates (recently extracted human teeth and acrylic prosthesis basis) were used to obtain the 96 test-specimens. Four adhesive systems (Cyanoacrylate ester, 3M Concise Enamel Bond Resin with or without previous enamel etching, MMA/HEMA or PMMA/MMA/ HEMA) were chosen for the fixation of the bioadhesives to substrate. Artificial saliva or distilled water was used as medium for maintaining the specimens until test. RESULTS: Statistical analysis showed that the interaction bioadhesives/acrylic prosthesis basis/cyanoacrylate ester adhesive was the most resistant to the physical removal by shearing. CONCLUSION: The newly rounded semi-convex format of acrylic device developed in this study presented satisfactory shear bond strength and might contribute to the comfort of intra-buccal use.
