The present paper reports the preparation and characterization of gelatin microspheres containing (a) a 44mer single-stranded synthetic oligonucleotide, complementary to the HLA-DRA gene (ssDNA-44) and (b) a double stranded fragment, 144 bp in length, prepared by polymerase chain reaction (PCR) mimicking a region of the HIV-1 LTR (dsDNA-144). Spherical gelatin microspheres were obtained by a coacervation method, showing high percentage of encapsulation yields (over 85%). Size distribution analysis of the produced microspheres results in an average diameter of 22 μm. In order to analyse the release profiles of both ssDNA-44 and dsDNA-144 from microspheres in vitro studies were carried out by using a flow through cell method. The chemical stability of dsDNA-144 to the encapsulation procedure steps was in addition demonstrated by PCR re-amplification of the DNA eluted from the gelatin microspheres. The reported results indicate that gelatin based microspheres offer excellent potential as carrier system for the in vivo administration of both single- and double-stranded DNA molecules.
GELATIN MICROSPHERES AS A NEW APPROACH FOR THE CONTROLLED DELIVERY OF SYNTHETIC OLIGONUCLEOTIDES AND PCR-GENERATED DNA FRAGMENTS
CORTESI, Rita;ESPOSITO, Elisabetta;MENEGATTI, Enea;GAMBARI, Roberto;NASTRUZZI, Claudio
1994
Abstract
The present paper reports the preparation and characterization of gelatin microspheres containing (a) a 44mer single-stranded synthetic oligonucleotide, complementary to the HLA-DRA gene (ssDNA-44) and (b) a double stranded fragment, 144 bp in length, prepared by polymerase chain reaction (PCR) mimicking a region of the HIV-1 LTR (dsDNA-144). Spherical gelatin microspheres were obtained by a coacervation method, showing high percentage of encapsulation yields (over 85%). Size distribution analysis of the produced microspheres results in an average diameter of 22 μm. In order to analyse the release profiles of both ssDNA-44 and dsDNA-144 from microspheres in vitro studies were carried out by using a flow through cell method. The chemical stability of dsDNA-144 to the encapsulation procedure steps was in addition demonstrated by PCR re-amplification of the DNA eluted from the gelatin microspheres. The reported results indicate that gelatin based microspheres offer excellent potential as carrier system for the in vivo administration of both single- and double-stranded DNA molecules.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
