title: Effect of chitosan malate on viability and cytoskeletal structures morphology of Caco-2 cells.
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 5mg/mL, respectively, while at 2.5mg/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 10mg 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 5-15mg/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.
authors: Dipartimento di Scienze del Farmaco, University of Sassari, Via Muroni 23/A, 07100 Sassari, Italy. julianoc@uniss.it
authors: Claudia
authors: C
authors: Juliano
authors: Grazia
authors: G
authors: Galleri
authors: Therese
authors: T
authors: Klemetsrud
authors: Jan
authors: J
authors: Karlsen
authors: Paolo
authors: P
authors: Giunchedi
mesh_terms: D000199:Actins / Q000378:metabolism; D018938:Caco-2 Cells; D002470:Cell Survival / Q000187:drug effects; D048271:Chitosan / Q000031:analogs & derivatives* / Q000633:toxicity; D004337:Drug Carriers; D006801:Humans; D008293:Malates / Q000633:toxicity*; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D019999:Pharmaceutical Solutions; D022502:Stress Fibers / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D014404:Tubulin / Q000378:metabolism
chemical_list: D000199:Actins; D004337:Drug Carriers; D008293:Malates; D019999:Pharmaceutical Solutions; D014404:Tubulin; C524437:chitosan malate; D048271:Chitosan