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Publications (3)3.42 Total impact

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    ABSTRACT: In this study, we explore effective in vivo plasmid and siRNA delivery vehicle in liver parenchymal cells especially in the hepatocytes. Recently developed biodegradable polymer, PolyJet based DNA transfection reagent that ensures effective and reproducible transfection on a broad ranges of hard-to-transfect mammalian cells, was selected to transfect hepatocytes. We compared PolyJet versus jetPEI and Lipofectamine in HEK293 cells and in primary culture of rat hepatocytes and Kupffer cells. We used TEX615 red dye, pmaxGFP and PGL3 plasmid to measure transfection efficiency. Our results indicated a significant and efficient translocation of the PolyJet conjugated pmaxGFP plasmid or dye labeled siRNA into hepatocytes, Kupffer cells and HEK293 cell lines compared to other transfection reagents. Our results also demonstrated efficient translocation of PGL3 plasmid into hepatocytes as measured by increase in luciferase activity. The transfection of PGL3 was improved through the Polyjet mediated approach as compared to that of Lipofectamine and jetPEI-Hepatocyte.
    Journal of Pharmaceutical Sciences and Pharmacology. 01/2014; 1(2).
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    ABSTRACT: Effective delivery of oligonucleotides to liver cells in vivo has been of significant interest in the field. Recently developed cationic polymer Polyethylenimine (PEI) with repeating ethylenimine motifs that provide a very high density of positive charges along its backbone have been reported as highly effective in siRNA delivery to target liver cells. This approach has been further optimized for hepatocyte specific delivery through galactose conjugation to target galactose-specific membrane lectins such as the asialoglycoprotein receptor. We compared jetPEI, jetPEI-GAL and jetPEI-Hepatocyte for their efficacy of delivering plasmids and dye labeled siRNA in cultured primary rat hepatocytes as well as in vivo in rat. Our results demonstrate that all three nanoparticle based approaches are effective at in vitro and in vivo delivery. We delivered PGL3 plasmid, FAM green dye and TEX615 red dye to measure fluorescence intensity, indicative of highly significant and efficient translocation of the plasmid or dye labeled siRNA into hepatocytes.
    Journal of Pharmaceutical Sciences and Pharmacology. 01/2014; 1(2).
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    ABSTRACT: BACKGROUND: Adaptation to chronic ethanol (EtOH) treatment of rats results in a changed functional state of the liver and greatly inhibits its regenerative ability, which may contribute to the progression of alcoholic liver disease. METHODS: In this study, we investigated the effect of chronic EtOH intake on hepatic microRNA (miRNA) expression in male Sprague-Dawley rats during the initial 24 hours of liver regeneration following 70% partial hepatectomy (PHx) using miRNA microarrays. miRNA expression during adaptation to EtOH was investigated using RT-qPCR. Nuclear factor kappa B (NFκB) binding at target miRNA promoters was investigated with chromatin immunoprecipitation. RESULTS: Unsupervised clustering of miRNA expression profiles suggested that miRNA expression was more affected by chronic EtOH feeding than by the acute challenge of liver regeneration after PHx. Several miRNAs that were significantly altered by chronic EtOH feeding, including miR-34a, miR-103, miR-107, and miR-122 have been reported to play a role in regulating hepatic metabolism and the onset of these miRNA changes occurred gradually during the time course of EtOH feeding. Chronic EtOH feeding also altered the dynamic miRNA profile during liver regeneration. Promoter analysis predicted a role for NFκB in the immediate-early miRNA response to PHx. NFκB binding at target miRNA promoters in the chronic EtOH-fed group was significantly altered and these changes directly correlated with the observed expression dynamics of the target miRNA. CONCLUSIONS: Chronic EtOH consumption alters the hepatic miRNA expression profile such that the response of the metabolism-associated miRNAs occurs during long-term adaptation to EtOH rather than as an acute transient response to EtOH metabolism. Additionally, the dynamic miRNA program during liver regeneration in response to PHx is altered in the chronically EtOH-fed liver and these differences reflect, in part, differences in miRNA expression between the EtOH-adapted and control livers at the baseline state prior to PHx.
    Alcoholism Clinical and Experimental Research 07/2012; · 3.42 Impact Factor