Intracellular delivery of liposome-encapsulated prolidase in cultured fibroblasts from prolidase-deficient patients

Paola Perugini, Khaolé Hassan, Ida Genta, Tiziana Modena, Franca Pavanetto, Giuseppe Cetta, Chiara Zanone, Paolo Iadarola, Annalia Asti, Bice Conti

Research output: Contribution to journalArticlepeer-review

Abstract

Prolidase is a cytosolic exopeptidase whose deficiency causes the development of a rare autosomal recessive disorder known as Prolidase Deficiency (PD). The main manifestations of PD are intractable ulcerations of the skin, recurrent infections and mental retardation. At this time only a hazardous and expensive chronic therapy based on blood transfusions is the suggested treatment for PD. The aim of this work was to investigate the capability of utilizing liposomes as enzyme carriers: these vesicular systems have been recently evaluated as protein carriers for their potential in terms of "in vivo" localization, drug release and for protein stabilization in biological fluids. Liposomes were prepared, with a 1:1 PC:Col molar ratio with or without DSPE-PEG, by a thin-film hydration. Ex-vivo experiments were performed, incubating prolidase loaded liposomes with cultured fibroblasts from PD patients and from controls, to determine the amount of active enzyme delivered to cells. Evaluation of liposomes toxicity on cultured skin fibroblasts showed that liposomes did not interfer with cellular growth. Results showed that all the active prolidase encapsulated in the liposomes was completely vehiculated inside fibroblasts after 6 days incubation. SEM analysis suggests that prolidase is vehiculated inside the cell through liposome endocytosis.

Original languageEnglish
Pages (from-to)181-190
Number of pages10
JournalJournal of Controlled Release
Volume102
Issue number1
DOIs
Publication statusPublished - Jan 20 2005

Keywords

  • Enzyme
  • Intracellular delivery
  • Liposome
  • Prolidase

ASJC Scopus subject areas

  • Pharmaceutical Science

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