caa a22 4500 445313374 CHVBK 20180317142638.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s11095-010-0345-x doi (NATIONALLICENCE)springer-10.1007/s11095-010-0345-x Uptake and Transport of Novel Amphiphilic Polyelectrolyte-Insulin Nanocomplexes by Caco-2 Cells—Towards Oral Insulin [Elektronische Daten] [Colin Thompson, Woei Cheng, Pramod Gadad, Karen Skene, Mhairi Smith, Gillian Smith, Alastair McKinnon, Rachel Knott] ABSTRACT: Purpose: The influence of polymer architecture on cellular uptake and transport across Caco-2 cells of novel amphiphilic polyelectrolyte-insulin nanocomplexes was investigated. Method: Polyallylamine (PAA) (15kDa) was grafted with palmitoyl chains (Pa) and subsequently modified with quaternary ammonium moieties (QPa). These two amphiphilic polyelectrolytes (APs) were tagged with rhodamine, and their uptake by Caco-2 cells or their polyelectrolyte complexes (PECs) with fluorescein isothiocyanate-insulin (FITC-insulin) uptake was investigated using fluorescence microscopy. The integrity of the monolayer was determined by measurement of transepithelial electrical resistance (TEER), and insulin transport across the monolayers was determined. Result: Pa and insulin were co-localised in cell membranes, while QPa complexes were found within the cytoplasm. QPa complex uptake was not affected by calcium, cytochalasin D or nocodazole. Uptake was reduced by co-incubation with sodium azide, an active transport inhibitor. Both polymers opened tight junctions reversibly, and insulin transport through monolayers increased when QPa or Pa was used. Conclusion: These APs have been shown to be taken up by Caco-2 cells and reversibly open tight cell junctions. Further work is required to optimise these formulations with a view to maximising their potential to facilitate oral delivery of insulin. Springer Science+Business Media, LLC, 2011 amphiphilic polyelectrolytes nationallicence Caco-2 nationallicence insulin nationallicence quaternary ammonium moiety nationallicence transepithelial electrical resistance nationallicence AP : amphiphilic polyelectrolyte nationallicence EMEM : eagle minimum essential media nationallicence Pa : palmitoyl nationallicence PAA : polyallylamine nationallicence PEC : polyelectrolyte complex nationallicence Q : quaternary ammonium moiety nationallicence TB : trypan blue nationallicence TEER : transepithelial electrical resistance nationallicence Thompson Colin School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut Cheng Woei School of Pharmacy, University of Hertfordshire, AL10 9AB, Hatfield, UK aut Gadad Pramod School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut Skene Karen School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut Smith Mhairi Histology and EM Facility, University of Aberdeen Institute of Medical Sciences, Foresterhill, AB25 2ZD, Aberdeen, UK aut Smith Gillian Histology and EM Facility, University of Aberdeen Institute of Medical Sciences, Foresterhill, AB25 2ZD, Aberdeen, UK aut McKinnon Alastair Histology and EM Facility, University of Aberdeen Institute of Medical Sciences, Foresterhill, AB25 2ZD, Aberdeen, UK aut Knott Rachel School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/4(2011-04-01), 886-896 0724-8741 28:4<886 2011 28 11095 https://doi.org/10.1007/s11095-010-0345-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-010-0345-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Thompson Colin School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut NATIONALLICENCE 700 1- Cheng Woei School of Pharmacy, University of Hertfordshire, AL10 9AB, Hatfield, UK aut NATIONALLICENCE 700 1- Gadad Pramod School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut NATIONALLICENCE 700 1- Skene Karen School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut NATIONALLICENCE 700 1- Smith Mhairi Histology and EM Facility, University of Aberdeen Institute of Medical Sciences, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 700 1- Smith Gillian Histology and EM Facility, University of Aberdeen Institute of Medical Sciences, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 700 1- McKinnon Alastair Histology and EM Facility, University of Aberdeen Institute of Medical Sciences, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 700 1- Knott Rachel School of Pharmacy and Life Sciences, The Robert Gordon University, Schoolhill, AB10 1FR, Aberdeen, UK aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/4(2011-04-01), 886-896 0724-8741 28:4<886 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer