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Cubosomes for topical delivery of the antimicrobial peptide LL-37

Boge, Lukas; Hallstensson, Karin; Ringstad, Lovisa; Johansson, Jenny; Andersson, Therese; Davoudi, Mina LU ; Larsson, Per Tomas; Mahlapuu, Margit; Håkansson, Joakim and Andersson, Martin (2019) In European Journal of Pharmaceutics and Biopharmaceutics 134. p.60-67
Abstract

In this study, the use of cubosomes for topical delivery of the antimicrobial peptide (AMP) LL-37 was investigated. Topical delivery of AMPs is of great interest for treatment of skin infections caused by bacteria, such as Staphylococcus aureus. AMP containing cubosomes were produced by three different preparation protocols and compared: (i) pre-loading, where LL-37 was incorporated into a liquid crystalline gel, which thereafter was dispersed into nanoparticles, (ii) post-loading, where LL-37 was let to adsorb onto pre-formed cubosomes, and (iii) hydrotrope-loading, where LL-37 was incorporated during the spontaneously formed cubosomes in an ethanol/glycerol monooleate mixture. Particle size and size distribution were analyzed using... (More)

In this study, the use of cubosomes for topical delivery of the antimicrobial peptide (AMP) LL-37 was investigated. Topical delivery of AMPs is of great interest for treatment of skin infections caused by bacteria, such as Staphylococcus aureus. AMP containing cubosomes were produced by three different preparation protocols and compared: (i) pre-loading, where LL-37 was incorporated into a liquid crystalline gel, which thereafter was dispersed into nanoparticles, (ii) post-loading, where LL-37 was let to adsorb onto pre-formed cubosomes, and (iii) hydrotrope-loading, where LL-37 was incorporated during the spontaneously formed cubosomes in an ethanol/glycerol monooleate mixture. Particle size and size distribution were analyzed using dynamic light scattering (DLS), liquid crystalline structure by small angle x-ray scattering (SAXS) and release of LL-37 by a fluorescamine assay. Proteolytic protection of LL-37 as well as bactericidal effect after enzyme exposure was investigated. The skin irritation potential of cubosomes was examined by an in vitro epidermis model. Finally, the bacterial killing property of the cubosomes was examined by an ex vivo pig skin wound infection model with Staphylococcus aureus. Data showed that a high loading of LL-37 induced formation of vesicles in case of cubosomes prepared by sonication (pre-loading). No release of LL-37 was observed from the cubosomes, indicating strong association of the peptide to the particles. Proteolysis studies showed that LL-37 was fully protected against enzymatic attacks while associated with the cubosomes, also denoting strong association of the peptide to the particles. As a consequence, bactericidal effect after enzyme exposure remained, compared to pure LL-37 which was subjected to proteolysis. No skin irritation potential of the cubosomes was found, thus enabling for topical administration. The ex vivo wound infection model showed that LL-37 in pre-loaded cubosomes killed bacteria most efficient.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Antimicrobial peptide, Cubosome, Infection, LL-37, Proteolysis, Skin irritation, Topical delivery
in
European Journal of Pharmaceutics and Biopharmaceutics
volume
134
pages
8 pages
publisher
Elsevier
external identifiers
  • scopus:85056876558
ISSN
0939-6411
DOI
10.1016/j.ejpb.2018.11.009
language
English
LU publication?
yes
id
e19a563c-691c-4646-88cc-4cd32436422f
date added to LUP
2018-12-03 09:49:00
date last changed
2018-12-04 03:00:11
@article{e19a563c-691c-4646-88cc-4cd32436422f,
  abstract     = {<p>In this study, the use of cubosomes for topical delivery of the antimicrobial peptide (AMP) LL-37 was investigated. Topical delivery of AMPs is of great interest for treatment of skin infections caused by bacteria, such as Staphylococcus aureus. AMP containing cubosomes were produced by three different preparation protocols and compared: (i) pre-loading, where LL-37 was incorporated into a liquid crystalline gel, which thereafter was dispersed into nanoparticles, (ii) post-loading, where LL-37 was let to adsorb onto pre-formed cubosomes, and (iii) hydrotrope-loading, where LL-37 was incorporated during the spontaneously formed cubosomes in an ethanol/glycerol monooleate mixture. Particle size and size distribution were analyzed using dynamic light scattering (DLS), liquid crystalline structure by small angle x-ray scattering (SAXS) and release of LL-37 by a fluorescamine assay. Proteolytic protection of LL-37 as well as bactericidal effect after enzyme exposure was investigated. The skin irritation potential of cubosomes was examined by an in vitro epidermis model. Finally, the bacterial killing property of the cubosomes was examined by an ex vivo pig skin wound infection model with Staphylococcus aureus. Data showed that a high loading of LL-37 induced formation of vesicles in case of cubosomes prepared by sonication (pre-loading). No release of LL-37 was observed from the cubosomes, indicating strong association of the peptide to the particles. Proteolysis studies showed that LL-37 was fully protected against enzymatic attacks while associated with the cubosomes, also denoting strong association of the peptide to the particles. As a consequence, bactericidal effect after enzyme exposure remained, compared to pure LL-37 which was subjected to proteolysis. No skin irritation potential of the cubosomes was found, thus enabling for topical administration. The ex vivo wound infection model showed that LL-37 in pre-loaded cubosomes killed bacteria most efficient.</p>},
  author       = {Boge, Lukas and Hallstensson, Karin and Ringstad, Lovisa and Johansson, Jenny and Andersson, Therese and Davoudi, Mina and Larsson, Per Tomas and Mahlapuu, Margit and Håkansson, Joakim and Andersson, Martin},
  issn         = {0939-6411},
  keyword      = {Antimicrobial peptide,Cubosome,Infection,LL-37,Proteolysis,Skin irritation,Topical delivery},
  language     = {eng},
  pages        = {60--67},
  publisher    = {Elsevier},
  series       = {European Journal of Pharmaceutics and Biopharmaceutics},
  title        = {Cubosomes for topical delivery of the antimicrobial peptide LL-37},
  url          = {http://dx.doi.org/10.1016/j.ejpb.2018.11.009},
  volume       = {134},
  year         = {2019},
}