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

Juliano, Claudia Clelia Assunta and Galleri, Grazia and Klemetsrud, Therese and Karlesen, Jan and Giunchedi, Paolo (2011) Effect of chitosan malate on viability and cytoskeletal structures morphology of Caco-2 cells. International Journal of Pharmaceutics, Vol. 420 (2), p. 223-230. ISSN 0378-5173. eISSN 1873-3476. Article.

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DOI: 10.1016/j.ijpharm.2011.08.039

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.
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.
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 in vivo after application of drug-loaded chitosan microparticles onto mucosal cells.

Item Type:Article
ID Code:7823
Status:Published
Refereed:Yes
Uncontrolled Keywords:Chitosan malate, Caco-2 cells, cytotoxicity, cell cytoskeleton Tubulin Actin
Subjects:Area 03 - Scienze chimiche > CHIM/09 Farmaceutico tecnologico applicativo
Divisions:002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
001 Università di Sassari > 01 Dipartimenti > Scienze del farmaco
Publisher:Elsevier
ISSN:0378-5173
eISSN:1873-3476
Deposited On:25 Jul 2012 09:22

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