Cationized pullulan 3D matrices as new materials for gene transfer.
This study deals with the development of a novel biocompatible cationized pullulan three-dimensional matrix for gene delivery. A water-soluble cationic polysaccharide, diethylaminoethyl-pullulan (DEAE-pullulan), was first synthesized and characterized. Fluorescence quenching and gel retardation assays evidenced the complexation in solution of DNA with DEAE-pullulan, but not with neutral pullulan. On cultured smooth muscle cells (SMCs) incubated with DEAE-pullulan and a plasmid vector expressing a secreted form of alkaline phosphatase (pSEAP), SEAP activity was 150-fold higher than with pSEAP alone or pSEAP with neutral pullulan. DEAE-pullulan was then chemically crosslinked using phosphorus oxychloride. The resulting matrices were obtained in less than a minute and molded as discs of 12 mm diameter and 2 mm thickness. Such DEAE-pullulan 3D matrices were loaded with up to 50 microg of plasmid DNA, with a homogeneous plasmid loading observed with YOYO-1 fluorescence staining. Moreover, the DEAE-pullulan matrix was shown to protect pSEAP from DNase I degradation. Incubation of cultured SMCs with pSEAP-loaded DEAE-pullulan matrices resulted in significant gene transfer without cell toxicity. This study suggests that these cationized pullulan 3D matrices could be useful biomaterials for local gene transfer.
San Juan
Aurélie
A
Inserm, U698, Bio-ingénierie Cardiovasculaire, Université Paris 7, Paris F-75018, Université Paris 13, Villetaneuse F-93430, France.
Hlawaty
Hanna
H
Chaubet
Frédéric
F
Letourneur
Didier
D
Feldman
Laurent J
LJ
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002412:Cations; D002478:Cells, Cultured; D004983:Ethanolamines; D018014:Gene Transfer Techniques; D005936:Glucans / Q000737:chemistry*; D008422:Materials Testing; D008969:Molecular Sequence Data; D015394:Molecular Structure; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D014162:Transfection
D001672:Biocompatible Materials; D002412:Cations; D004983:Ethanolamines; D005936:Glucans; C009109:pullulan; C007369:2-diethylaminoethanol