High content analysis of cytotoxic effects of pDMAEMA on human intestinal epithelial and monocyte cultures

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Title: High content analysis of cytotoxic effects of pDMAEMA on human intestinal epithelial and monocyte cultures
Authors: Rawlinson, Lee-Anne Betty
O'Brien, Peter J.
Brayden, David James
Permanent link: http://hdl.handle.net/10197/3009
Date: Aug-2010
Online since: 2011-07-11T15:32:45Z
Abstract: Poly(2-(dimethylamino ethyl)methacrylate) (pDMAEMA) is a cationic polymer with potential as an antimicrobial agent and as a non-viral gene delivery vector. The aim was to further elucidate the cytotoxicity of a selected pDMAEMA low molecular weight (MW) polymer against human U937 monocytes and Caco-2 intestinal epithelial cells using a novel multi-parameter high content analysis (HCA) assay and to investigate histological effects on isolated rat intestinal mucosae. Seven parameters of cytotoxicity were measured: nuclear intensity (NI), nuclear area (NA), intracellular calcium ([Ca2+]i), mitochondrial membrane potential (MMP), plasma membrane permeability (PMP), cell number (CN) and phospholipidosis. Histological effects of pDMAEMA on excised rat ileal and colonic mucosae were investigated in Ussing chambers. Following 24-72 hours exposure, 25-50 µg/ml pDMAEMA induced necrosis in U937 cells, while 100-250 µg/ml induced apoptosis in Caco-2. pDMAEMA increased NA and NI and decreased [Ca2+]i, PMP, MMP and CN in U937 cells. In Caco-2, it increased NI and [Ca2+]i, but decreased NA, PMP, MMP and CN. Phospholipidosis was not observed in either cell line. pDMAEMA (10 mg/ml) did not induce any histological damage on rat colonic tissue and only mild damage to ileal tissue following exposure for 60 min. In conclusion, HCA reveals that pDMAEMA induces cytotoxicity in different ways on different cell types at different concentrations. HCA potential for high throughput toxicity screening in drug formulation programmes.
Funding Details: Science Foundation Ireland
Type of material: Journal Article
Publisher: Elsevier
Journal: Journal of Controlled Release
Volume: 146
Issue: 1
Start page: 84
End page: 92
Copyright (published version): 2010 Elsevier B.V.
Keywords: High content analysisCytotoxicity assayspDMAEMAAntimicrobial polymersCaco-2 intestinal epithelia
Subject LCSH: Polymers in medicine
Anti-infective agents
Biological assay
DOI: 10.1016/j.jconrel.2010.05.002
Other versions: http://dx.doi.org/10.1016/j.jconrel.2010.05.002
Language: en
Status of Item: Peer reviewed
Appears in Collections:Irish Drug Delivery Network Research Collection

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