Study on the toxicity and transfection efficiency of PEI/MYOC gene complexes in vivo / 眼科

ABSTRACT

Objective To investigate the toxicity and transfection efficiency of PEI-mediated plasmid DNA encoding myocilin (MYOC) gene transfected into trabecular meshwork.Design Experimental study.Participants SD rats.Methods PEI and plasmid encoding MYOC gene complex was prepared and 20?l PEI/DNA complex was directly injected into anterior chamber of SD rat by microinjection. The eyes were extracted at day 1,3,7,and 14 after injection for the structure analysis of trabecular meshwork by HE staining and transmission electron microscope.Expression of the transferred myocilin gene was monitored by fluorescence microscopy. Main Outcome Measures HE staining,immunohistochemistry and electron microscope.Results No significant toxicity or inflammation was detected under the HE staining and electron microscope.PEI/DNA complex were seen in cytoplasm of trabecular meshwork cells. The fluorescence intensity of myocilin expression in the trabecular meshwork cells was increasing with time and peaked on clays 3 and declined afterward.Conclusion Plasmid DNA mediated by PEI could be efficiently transfected into trabecular meshwork cells by directly injecting into anterior chamber without any toxicity.