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Effect of chitosan malate on viability and cytoskeletal structures morphology of Caco-2 cells

By Claudia Clelia Assunta Juliano, Grazia Galleri, Therese Klemetsrud, Jan Karlesen and Paolo Giunchedi

Abstract

The aim of this work was to evaluate the effects of the treatment with chitosan malate (CM) on viability of Caco-2 cells and on the morphology and the integrity of their cytoskeletal structures (microtubules, microfilaments). Cytotoxicity of CM, both as a solution and as microparticles obtained by spray drying, was evaluated by using the reduction of MTT reagent; microtubule and microfilaments organization of Caco-2 cells treated with CM solution was examined with immunofluorescence techniques in monolayers fixed with the glutaraldehyde–borohydride method. CM as a solution displayed a concentration-dependent cytotoxicity towards Caco-2 cells, with viability percentages of 5 ± 2%, 7 ± 3% and 31 ± 15% at 15, 10 and 5 mg/mL, respectively, while at 2.5 mg/mL or less cell viability was 90% or higher. CM microparticles also produced a remarkable cytotoxic effect (cell viability 84 ± 17%, 16 ± 8% and 5 ± 6% after treatment with 1, 5 and 10 mg CM per well, respectively), resulting more toxic than CM solution. <br>Microtubules pattern of Caco-2 cells, which is a network regularly arranged around the nucleus, appeared deeply modified by CM treatment in a concentration-dependent way, with progressive microtubule changes in length and spatial disposition and deposition of fluorescent aggregates at the periphery of the cells. Furthermore, after treatment with 5–15 mg/mL CM, remarkable alterations of actin organization were observed, with a progressive disruption of the network of stress fibers and the appearance of actin aggregates inside the cell cytoplasm.</br> In conclusion, viability and the cytoskeletal pattern of Caco-2 cells are modified by treatment with CM at high concentrations, which might be locally reached <i>in vivo</i> after application of drug-loaded chitosan microparticles onto mucosal cells

Topics: CHIM/09 Farmaceutico tecnologico applicativo
Publisher: Elsevier
Year: 2011
DOI identifier: 10.1016/j.ijpharm.2011.08.039
OAI identifier: oai:eprints.uniss.it:7823
Provided by: UnissResearch
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