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• W2313308425 abstract "Event Abstract Back to Event Hyaluronan-modification boosts the nanoparticle uptake by trabecular meshwork cells Michaela Guter1*, Renate Liebl1 and Miriam Breunig1 1 University of Regensburg, Department of Pharmaceutical Technology, Faculty of Chemistry and Pharmacy, Germany Introduction: Glaucoma is one of the leading causes of blindness worldwide[1]. Nanoparticles targeting the trabecular meshwork (TM) could potentially overcome the limitations of current therapies[2]. We propose applying hyaluronan (HA) modified nanoparticles (NP) for TM targeting. The NP are envisioned to interact with cell-surface CD44 receptors present on TM cells. In this study, we determined the CD44-expression on TM cells to prove the feasibility of our approach. Furthermore, we investigated the influence of the HA-coating of NP on their cellular uptake and endosomal escape. Materials and Methods: NP were prepared by nanoprecipitation by using fluorescently labeled polylactid-glycolid (PLGA) 30-40 kDa (Polyscitech) and polyethyleneimine (PEI) 25 kDa branched (Sigma) as a stabilizing agent. Functionalization with HA 13 kDa (Lifecore) was performed in a layer-by-layer approach[3]. Extent of CD44-expression was determined by immunostaining using AlexaFluor®488 anti-CD44 antibody (Clone IM7, BioLegend) on a FACS calibur. For uptake experiments, cells were incubated with NP after receptor blocking with HA 289 kDa (Lifecore). Intracellular fate of NP was visualized by calcein-assay[4] on a Zeiss Laser Scanning Microscope. Results and Discussion: Immortalized human trabecular meshwork cells (HTM) were compared with two cell lines of low (MCF-7) and high (MDA-MB-231) CD44 expression level. The receptor density increased in the following order: MCF-7 < HTM < MDA-MB-231 providing a good basis for NP interaction with HTM cells. To investigate the influence of HA-coating on cellular uptake, HA modified NPs (PLGA-PEI/HA, d =175 nm, ζ = -28 mV) and unmodified ones (PLGA-PEI, d = 198 nm, ζ = +53 mV) were analyzed (Figure 1). As expected, nanoparticle uptake by CD44 receptor positive MDA-MB-231 and HTM was significantly higher after functionalization with HA (p=0.002 or p<0.001 respectively). In the case of MCF-7, which exhibited negligible CD44 expression, no difference between modified and unmodified NP was detected. In addition, pretreatment with high molecular weight HA demonstrated at least a partial receptor-mediated uptake. Interestingly, a reduction of nanoparticle uptake was also observed for PLGA-PEI. An essential step for drug delivery is the intracellular endosomal escape. Figure 2 nicely shows that a HA coating resulted in a higher endosomal escape capacity of the NP compared to the unmodified ones. Conclusion: Modification of NP with HA is a successful approach to improve their cellular uptake and endosomal escape in HTM cells. These nanoparticles might become a promising tool for drug delivery in glaucoma therapy. References:[1] World Health Organization, 2012, Global Data on visual impairments 2010[2] Pita-Thomas, Daniel W.; Goldberg, Jeffrey L.; Nanotechnology and glaucoma: little particles for a big disease. Curr. Opin. Ophthalmol. 2013, 24, 130-135[3] Elbakry, Asmaa; Wurster, Eva-Christina; Zaky, Alaa; Liebl, Renate; Schindler, Edith; Bauer-Kreisel, Petra; Blunk, Torsten; Rachel, Reinhard; Goepferich, Achim; Breunig, Miriam; Layer-by-Layer Coated Gold Nanoparticles: Size-Dependent Delivery of DNA into Cells. Small 2012, 8, 3847-56[4] Bonner, Daniel K.; Leung, Cheuk; Chen-Liang, Jane; Chingozha, Loice; Langer, Robert; Hammond, Paula T.; Intracellular Trafficking of Polyamidoamine-Poly(ethylene glycol) Block Copolymers in DNA Delivery. Bioconjugate Chem. 2011, 22, 1519-1525 Keywords: nanoparticle, Polymeric material, targeting delivery, Cell interaction Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Tissue targeting nanoparticles Citation: Guter M, Liebl R and Breunig M (2016). Hyaluronan-modification boosts the nanoparticle uptake by trabecular meshwork cells. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.01058 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. * Correspondence: Dr. Michaela Guter, University of Regensburg, Department of Pharmaceutical Technology, Faculty of Chemistry and Pharmacy, Regensburg, Germany, Email1 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 Michaela Guter Renate Liebl Miriam Breunig Google Michaela Guter Renate Liebl Miriam Breunig Google Scholar Michaela Guter Renate Liebl Miriam Breunig PubMed Michaela Guter Renate Liebl Miriam Breunig Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page." @default.
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• W2313308425 title "Hyaluronan-modification boosts the nanoparticle uptake by trabecular meshwork cells" @default.
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