title: Reduced cytotoxicity and enhanced bioactivity of cationic antimicrobial peptides liposomes in cell cultures and 3D epidermis model against HSV.
abstract: Cationic antimicrobial peptides (AMPs) are part of the innate immunity, and act against a wide variety of pathogenic microorganisms by perturbation of the microorganism's plasma membrane. Although attractive for clinical applications, these agents suffer from limited stability and activity in vivo, as well as non-specific interaction with host biological membranes, leading to cytotoxic adverse effects. We hypothesized that encapsulation of AMPs within liposomes could result in reduced cytotoxicity, and with enhanced stability as well as bioactivity against herpes simplex virus 1 (HSV-1). We formulated nano-sized liposomal formulations of LL-37 and indolicidin, and their physicochemical properties, cellular uptake, in vitro cytotoxicity and antiviral efficacy have been determined. Lower cytotoxicity of LL-37 liposomes was found in comparison to indolicidin liposomes attributed to the superior physicochemical properties, and to the different degree of interaction with the liposomal membrane. The disc-like shaped LL-37 liposomes (106.8±10.1nm, shelf-life stability of >1year) were taken up more rapidly and to a significantly higher extent than the free peptide by human keratinocyte cell line (HaCaT), remained intact within the cells, followed by release of the active peptide within the cytoplasm and migration of the vesicles' lipids to the plasma membrane. LL-37 liposomes were found significantly less toxic than both the free agent and liposomal indolicidin. In the new 3D epidermis model (immortalized primary keratinocytes) liposomal LL-37 treatment (>20μM), but not free LL-37, efficiently protected the epidermis, inhibiting HSV-1 infection. This positive antiviral effect was obtained with no cytotoxicity even at very high concentrations (400μM). Thus, the antiviral activity of encapsulated LL-37 was significantly improved, expanding its therapeutic window. Liposomal LL-37 appears to be a promising delivery system for HSV therapy.
authors.affiliation: Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Israel.
authors.forename: Sapir
authors.initials: S
authors.lastname: Ron-Doitch
authors.affiliation: Fraunhofer IGB, Stuttgart, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Sawodny
authors.affiliation: Fraunhofer IGB, Stuttgart, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kühbacher
authors.affiliation: Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Israel.
authors.forename: Mirjam M Nordling
authors.initials: MMN
authors.lastname: David
authors.affiliation: Linköping University, Linköping, Sweden.
authors.forename: Ayan
authors.initials: A
authors.lastname: Samanta
authors.affiliation: Linköping University, Linköping, Sweden.
authors.forename: Jaywant
authors.initials: J
authors.lastname: Phopase
authors.affiliation: Fraunhofer IGB, Stuttgart, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Burger-Kentischer
authors.affiliation: Linköping University, Linköping, Sweden.
authors.forename: May
authors.initials: M
authors.lastname: Griffith
authors.affiliation: Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Israel. Electronic address: gershong@ekmd.huji.ac.il.
authors.forename: Gershon
authors.initials: G
authors.lastname: Golomb
authors.affiliation: Fraunhofer IGB, Stuttgart, Germany.
authors.forename: Steffen
authors.initials: S
authors.lastname: Rupp
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000008:administration & dosage*; D000998:Antiviral Agents / Q000008:administration & dosage*; D018929:Cell Culture Techniques; D002460:Cell Line; D002478:Cells, Cultured; D004817:Epidermis / Q000821:virology; D052816:Foreskin / Q000166:cytology; D018259:Herpesvirus 1, Human / Q000187:drug effects*; D006801:Humans; D015603:Keratinocytes / Q000821:virology; D008055:Lipids / Q000737:chemistry; D008081:Liposomes; D008297:Male; D054804:Cathelicidins
chemical_list: D023181:Antimicrobial Cationic Peptides; D000998:Antiviral Agents; D008055:Lipids; D008081:Liposomes; C073143:indolicidin; D054804:Cathelicidins
keywords: 3D epidermis model; Antimicrobial peptides; Disc-like liposomes; Drug delivery system; HSV-1; Indolicidin; LL-37; Liposomes; Peptide delivery