Liposomes are considered very promising delivery systems for antisense therapeutic approach, offering drug protection and facilitating oligonucleotide cell internalization. The present study was aimed to investigate the influence of phospholipid composition of the liposomal systems both on the encapsulation and on the oligonucleotide carrier capacity in vitro. Liposomes composed of neutral ( phosphatidylcholine, cholesterol and dioleoylphosphatidylethanolamine) and/or cationic lipids (N-(1-(2,3-dioleoyloxy) propyl)-N,N,N-trimethylammonium chloride salt, DOTAP) with different molar ratios were complexed with 50 fluorescein conjugated 29-mer phosphorothioate oligonucleotide (PS-ODN). The interaction was evaluated using atomic force microscopy (AFM), gel electrophoresis and HPLC analysis. Cytofluorimetric analysis and fluorescence microscopy were applied to evaluate the uptake and intracellular distribution of fluorescently labelled PS-ODN after transfection in two cell lines, COS I ( fibroblast cell) and HaCaT ( immortalized keratinocyte cell). The AFM studies reveal that the liposome/PS-ODN interaction leads the formation of a new irregular structure that completely hides the PS-ODN. Gel electrophoresis experiments and HPLC analysis have clearly demonstrated that also neutral liposomes are able to keep a little amount of PS-ODN but without strain to the complexation; the interaction was weak and rapidly destabilized when the complex was added to the cells. Transfection experiments performed with different incubation times show that DOTAP liposomes increase the rate of cellular uptake of PS-ODN and seem to influence its intracellular distribution in COS I cells where the oligonucleotide looks localized in nucleoli. Similar behaviour, at a lesser extent, is exhibited in HaCaT cells.

Liposome-oligonucleotides interaction for in vitro uptake by COSI and HaCaT cells / Ruozi, Barbara; Battini, Renata; Tosi, Giovanni; Forni, Flavio; Vandelli, Maria Angela. - In: JOURNAL OF DRUG TARGETING. - ISSN 1061-186X. - STAMPA. - 13:(2005), pp. 295-304.

Liposome-oligonucleotides interaction for in vitro uptake by COSI and HaCaT cells

RUOZI, Barbara;BATTINI, Renata;TOSI, Giovanni;FORNI, Flavio;VANDELLI, Maria Angela
2005-01-01

Abstract

Liposomes are considered very promising delivery systems for antisense therapeutic approach, offering drug protection and facilitating oligonucleotide cell internalization. The present study was aimed to investigate the influence of phospholipid composition of the liposomal systems both on the encapsulation and on the oligonucleotide carrier capacity in vitro. Liposomes composed of neutral ( phosphatidylcholine, cholesterol and dioleoylphosphatidylethanolamine) and/or cationic lipids (N-(1-(2,3-dioleoyloxy) propyl)-N,N,N-trimethylammonium chloride salt, DOTAP) with different molar ratios were complexed with 50 fluorescein conjugated 29-mer phosphorothioate oligonucleotide (PS-ODN). The interaction was evaluated using atomic force microscopy (AFM), gel electrophoresis and HPLC analysis. Cytofluorimetric analysis and fluorescence microscopy were applied to evaluate the uptake and intracellular distribution of fluorescently labelled PS-ODN after transfection in two cell lines, COS I ( fibroblast cell) and HaCaT ( immortalized keratinocyte cell). The AFM studies reveal that the liposome/PS-ODN interaction leads the formation of a new irregular structure that completely hides the PS-ODN. Gel electrophoresis experiments and HPLC analysis have clearly demonstrated that also neutral liposomes are able to keep a little amount of PS-ODN but without strain to the complexation; the interaction was weak and rapidly destabilized when the complex was added to the cells. Transfection experiments performed with different incubation times show that DOTAP liposomes increase the rate of cellular uptake of PS-ODN and seem to influence its intracellular distribution in COS I cells where the oligonucleotide looks localized in nucleoli. Similar behaviour, at a lesser extent, is exhibited in HaCaT cells.
13
295
304
Liposome-oligonucleotides interaction for in vitro uptake by COSI and HaCaT cells / Ruozi, Barbara; Battini, Renata; Tosi, Giovanni; Forni, Flavio; Vandelli, Maria Angela. - In: JOURNAL OF DRUG TARGETING. - ISSN 1061-186X. - STAMPA. - 13:(2005), pp. 295-304.
Ruozi, Barbara; Battini, Renata; Tosi, Giovanni; Forni, Flavio; Vandelli, Maria Angela
File in questo prodotto:
File Dimensione Formato  
Articolo Ruozi et al-J Drug Targeting 2005.pdf

non disponibili

Tipologia: Versione dell'autore revisionata e accettata per la pubblicazione
Dimensione 7.54 MB
Formato Adobe PDF
7.54 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

Licenza Creative Commons
I metadati presenti in IRIS UNIMORE sono rilasciati con licenza Creative Commons CC0 1.0 Universal, mentre i file delle pubblicazioni sono rilasciati con licenza Attribuzione 4.0 Internazionale (CC BY 4.0), salvo diversa indicazione.
In caso di violazione di copyright, contattare Supporto Iris

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/310671
Citazioni
  • ???jsp.display-item.citation.pmc??? 2
  • Scopus 8
  • ???jsp.display-item.citation.isi??? 9
social impact