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

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 interfere 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.
AuthorsPaola Perugini, Khaolé Hassan, Ida Genta, Tiziana Modena, Franca Pavanetto, Giuseppe Cetta, Chiara Zanone, Paolo Iadarola, Annalia Asti, Bice Conti
JournalJournal of controlled release : official journal of the Controlled Release Society (J Control Release) Vol. 102 Issue 1 Pg. 181-90 (Jan 20 2005) ISSN: 0168-3659 [Print] Netherlands
PMID15653144 (Publication Type: Comparative Study, Journal Article)
Chemical References
  • Liposomes
  • Dipeptidases
  • proline dipeptidase
Topics
  • Cells, Cultured
  • Dipeptidases (deficiency, therapeutic use)
  • Drug Delivery Systems (methods)
  • Endocytosis
  • Fibroblasts (drug effects, enzymology)
  • Humans
  • Intracellular Fluid (drug effects, enzymology)
  • Liposomes

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