摘要
D-Aminoacid oxidase (DAO) was isolated from fresh porcine kidney;its cytotoxic potential was studied under in vitro and in vivo conditions. The isolated DAO was complexed with Fe2O3 nanoparticles and its potential as an oxidation therapeutic agent was analysed. The ability of the complex in eliciting H2O2 mediated cytotoxicity was studied on Dalton’s lymphoma ascites cells (DLA). The induction of apoptosis in DLA cells by Fe2O3-DAO complex was studied by morphological examination and alkaline single cell gel electrophoresis (comet assay). The antitumor activity of the complex was investigated by oral administration of the complex and the substrate D-alanine to tumor bearing Swiss albino mice and by targeting the complex to the tumor site, using an externally applied magnetic field. Fe2O3-DAO along with D-alanine showed remarkable cytotoxicity in a substrate concentration-dependent manner. Both morphological examination and comet assay revealed that Fe2O3-DAO/D-alanine induced apoptosis. Oral administration of Fe2O3-DAO and D-alanine along with magnetic targeting significantly suppressed tumor growth in mice. The present report provides the first evidence for the promising application of enzyme bound nanoparticles for targeted oxidation therapy.
D-Aminoacid oxidase (DAO) was isolated from fresh porcine kidney;its cytotoxic potential was studied under in vitro and in vivo conditions. The isolated DAO was complexed with Fe2O3 nanoparticles and its potential as an oxidation therapeutic agent was analysed. The ability of the complex in eliciting H2O2 mediated cytotoxicity was studied on Dalton’s lymphoma ascites cells (DLA). The induction of apoptosis in DLA cells by Fe2O3-DAO complex was studied by morphological examination and alkaline single cell gel electrophoresis (comet assay). The antitumor activity of the complex was investigated by oral administration of the complex and the substrate D-alanine to tumor bearing Swiss albino mice and by targeting the complex to the tumor site, using an externally applied magnetic field. Fe2O3-DAO along with D-alanine showed remarkable cytotoxicity in a substrate concentration-dependent manner. Both morphological examination and comet assay revealed that Fe2O3-DAO/D-alanine induced apoptosis. Oral administration of Fe2O3-DAO and D-alanine along with magnetic targeting significantly suppressed tumor growth in mice. The present report provides the first evidence for the promising application of enzyme bound nanoparticles for targeted oxidation therapy.
