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• W2321839538 abstract "Event Abstract Back to Event Pullulan based multifunctional thiolated polymers for efficient gene delivery and efflux pump inhibition with enhanced doxorubicin retention in c6 glioma cell Priya S. Syamala1 and Rekha M. Ramesan1 1 Sree Chitra Tirunal Institute for Medical Sciences and Technology, Division of Biosurface Technology, India Introduction: One of the major impediments to successful cancer chemotherapy is multidrug resistance attributed to P-glycoprotein (P-gp)[1]. Still, there is no real strategy for overcoming drug resistance in cancer.Thiolated polymers (thiomer) are known for its inherent P-gp inhibiting property[2]. In this study, pullulan based thiolated cationic polymer was synthesized by conjugating pullulan and branched chain polyethylenimine (25kda) with subsequent incorporation of disulphide linkage via introduction of mercaptosuccinic acid (PPMSS).The aim is to form pullulan based thiolated cationic polymer to simultaneously act as therapeutic gene delivery vehicle and inhibit P-glycoprotein to enhance DOX retention and drug mediated cytotoxicity in the tumour cells. Materials: Pullulan (MW75000Da; Fluka), Polyethyleneimine (PEI; MW 25,000 Da), Carbonyldiimidazole (CDI), EDC, Mercaptosuccinic acid were purchased from Sigma–Aldrich Chemicals Co, USA. Methods: The thiolated polymers were characterised in terms of size, surface charge, morphology and surface chemistry by DLS, TEM, NMR and Raman Spectroscopy. Endo osmolytic properties as well as reduction sensitivity of the polymer in presence of dithiothreitol (DTT) were evaluated. In vitro cytotoxicity, uptake and transfection efficiency along with anticancer effects such as efflux pump inhibition, DOX retention and drug mediated cytotoxicity were evaluated in C6 glioma cells in terms of microplate reader, flow cytometry and confocal image analysis. Results and discussion: The thiolated polymer,PPMSS/pDNA nanoplex formed particle size (150 – 200 nm) and surface charge (+15 to +20 mV) suitable for gene delivery vehicle. Interestingly, it showed significant endo osmolytic property and formed large aggregates in response to 25mM dithiothreitol(mimicking intracellular GSH) as shown by DLS and reductive cleavage of intermediate disulphide linkage resulted in the subsequent release of pDNA as evidenced in agarose gel electrophoresis, implies its potential as gene delivery vehicle.The thiolated polymer showed lower cytotoxicity and the PPMSS/pDNA nanoplexes exhibited high cellular uptake and improved transfection efficiency compared to PEI-MSA alone,confirming its role as an ideal gene delivery vector.Similarly, the P-gp inhibitory activity of PPMSS has also been revealed by significant efflux pump inhibition of nanoplex in C6 cell lines and associated retention of anticancer drug, doxorubicin (DOX) and drug mediated cytotoxicity in tumour cells. Figure 1: The confocal microscopic images of A and D) PPMSS/ YOYO labelled pDNA nanoplexes with polymer/DNA ratio 3:1 and 4:1 respectively in C6 cells. B& E) Hoechst 33342 stained nucleus C & F) merged confocal fluorescent images of YOYO tagged DNA & Hoechst stained nucleus of PPMSS/pDNA of ratios 3:1 and 4:1 respectively. Figure 2: DOX retention after preincubation with nanoplexes (PPMSS/DNA) for 2hrs and then addition of free DOX at different time periods, the control is medium and DOX alone. Conclusion: In this study, we developed pullulan based cationic polymer (PPMSS) to enable therapeutic gene delivery and efflux pump inhibition to improve chemotherapy in C6 glioma cell as evidenced by in vitro cytotoxicity,flow cytometry analysis and confocal imaging.These findings imply that PPMSS can be a promising P-gp inhibitor and a gene delivery vehicle for overcoming cancer drug resistance and mediate cancer gene therapy. References:[1] Krishna,R.; Mayer,L.W. Eur J Pharm Sci 2000,11, 265–283.[2] Rahmata,D.; Sakloetsakun,D. Shahnaz,G.; Sarti,F.; Laffleur,F.; Bernkop-Schnurch,A. Int J Pharm 2012,422, 40– 4 Keywords: Drug delivery, nanoparticle, gene delivery, Non-viral vector Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: General Session Oral Topic: Biomaterials in gene therapy Citation: Syamala PS and Ramesan RM (2016). Pullulan based multifunctional thiolated polymers for efficient gene delivery and efflux pump inhibition with enhanced doxorubicin retention in c6 glioma cell. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02403 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Priya S Syamala Rekha M Ramesan Google Priya S Syamala Rekha M Ramesan Google Scholar Priya S Syamala Rekha M Ramesan PubMed Priya S Syamala Rekha M Ramesan Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. 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• W2321839538 title "Pullulan based multifunctional thiolated polymers for efficient gene delivery and efflux pump inhibition with enhanced doxorubicin retention in c6 glioma cell" @default.
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