CATIONIC NANOEMULSIONS AS DELIVERY SYSTEMS FOR THE DNA
PLASMID pIDUA: PHYSICOCHEMICAL PROPERTIES, COMPLEXATION
AND STABILITY STUDIES
FRAGA, M1,2, MATTE, U2,3, TEIXEIRA, H.F.1
[email protected]
1
Post Graduation Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul
(UFRGS), Porto Alegre, RS, Brazil.
2
Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA),
Porto Alegre, RS, Brazil.
3
Post Graduation Program in Genetics and Molecular Biology, Universidade Federal do Rio Grande do
Sul (UFRGS), Porto Alegre, RS, Brazil.
Keywords: plasmids, cationic nanoemulsions, physicochemical characterization, gene transfer, DOTAP
1. Introduction
3. Results
Cationic nanoemulsions have been recently
The
considered as a potential delivery system for
yielded
DNA plasmids (pDNA) (1). Therefore, the aim
droplet size and ζ-potential of about 250 nm and
of
evaluate
+50 mV, respectively. The complexation of
physicochemical properties, complexation and
pIDUA with the cationic nanoemulsion was
stability of a cationic nanoemulsion complexed
total when the complex possesses a charge
with pIDUA. This pDNA contains the gene that
relation [+/-] ≥ 2.0. In these conditions, no
encodes for the enzyme alpha-L-iduronidase
significant differences of the oil droplet size of
(IDUA) which is necessary for the treatment of
nanoemulsions were detected after pIDUA
the genetic disorder Mucopolysaccharidosis
complexation by PCS and TEM. In these
type I.
conditions, the complex was protected from
2. Methods
enzymatic degradation by DNase I enzyme. For
A cationic nanoemulsion composed of medium
the control formulation without cationic lipid no
chain triglycerides, egg lecithin, cationic lipid
complex was formed and pDNA was degraded
DOTAP, glycerol and water was prepared
by DNase I enzyme.
the
present
work
was
to
spontaneous
emulsification
monodisperse
procedure
nanoemulsions
with
through spontaneous emulsification process (2).
It was also prepared a nanoemulsion without
4. Conclusion
cationic lipid as a control. The droplet size was
The overall results demonstrate the potential of
observed by means of photon correlation
cationic nanoemulsions as a transfer system for
spectroscopy (PCS) and transmission electron
the plasmid pIDUA.
microscopy (TEM) using uranyl acetate. ζ-
Acknowledgments
potential of nanoemulsions was determined
FIPE-HCPA, FAPERGS/CNPq
through
electrophoretic
mobility.
The
complexation of pDNA with the nanoemulsions
was analyzed by agarose gel retardation assay
and the stability was performed incubating the
complexes with 1 U DNase I enzyme for 30
minutes.
References
1. Verissimo LM, Lima LF, Egito LC, de Oliveira AG, do
Egito ES. Pharmaceutical emulsions: a new approach for
gene therapy. J Drug Target. 2010;18(5):333-342.
2. Fraga M, Laux M, Dos Santos GR, et al. Evaluation of the
toxicity
of
oligonucleotide/cationic
nanoemulsion
complexes on Hep G2 cells through MTT assay. Pharmazie.
2008;63(9):667-670.