Three-dimensional single-particle tracking (SPT) was used to calculate the mean square displacement (MSD) and the diffusion coefficients of multicomponent cationic liposome–DNA complexes (lipoplexes) in CHO-K1 living cells. In untreated (NT) control cells, we found that the intracellular lipoplex motion was either directed or Brownian with active transportation being definitely more frequent (more than 70%) than Brownian diffusion. The MSD analysis was supported by the calculation of the three-dimensional asphericity, A3, which was close to unity, denoting the preponderant occurrence of movement along a direction. To elucidate the role of the cytoskeleton structure in the lipoplex trafficking, cells were treated with cytoskeleton (actin microfilaments and microtubules) polymerization inhibitors (latrunculin B and nocodazole, respectively). When cells were treated with inhibitors, the lipoplex movement tended towards a random walk at the expense of directed motion. The disassembly of microtubules had a stronger effect on the reduction of directional movement than that of actin microfilaments. Relevance of the results for enhanced gene delivery is discussed.

Intracellular trafficking of cationic liposome–DNA complexes in living cells / Coppola, Stefano; Laura C., Estrada; Michelle A., Digman; Pozzi, Daniela; Francesco, Cardarelli; Enrico, Gratton; Caracciolo, Giulio. - In: SOFT MATTER. - ISSN 1744-6848. - ELETTRONICO. - 8:30(2012), pp. 7919-7927. [10.1039/c2sm25532d]

Intracellular trafficking of cationic liposome–DNA complexes in living cells

COPPOLA, STEFANO;POZZI, DANIELA;CARACCIOLO, Giulio
2012

Abstract

Three-dimensional single-particle tracking (SPT) was used to calculate the mean square displacement (MSD) and the diffusion coefficients of multicomponent cationic liposome–DNA complexes (lipoplexes) in CHO-K1 living cells. In untreated (NT) control cells, we found that the intracellular lipoplex motion was either directed or Brownian with active transportation being definitely more frequent (more than 70%) than Brownian diffusion. The MSD analysis was supported by the calculation of the three-dimensional asphericity, A3, which was close to unity, denoting the preponderant occurrence of movement along a direction. To elucidate the role of the cytoskeleton structure in the lipoplex trafficking, cells were treated with cytoskeleton (actin microfilaments and microtubules) polymerization inhibitors (latrunculin B and nocodazole, respectively). When cells were treated with inhibitors, the lipoplex movement tended towards a random walk at the expense of directed motion. The disassembly of microtubules had a stronger effect on the reduction of directional movement than that of actin microfilaments. Relevance of the results for enhanced gene delivery is discussed.
2012
lipoplexes; cationic liposomes; intracellular trafficking; dna; gene delivery
01 Pubblicazione su rivista::01a Articolo in rivista
Intracellular trafficking of cationic liposome–DNA complexes in living cells / Coppola, Stefano; Laura C., Estrada; Michelle A., Digman; Pozzi, Daniela; Francesco, Cardarelli; Enrico, Gratton; Caracciolo, Giulio. - In: SOFT MATTER. - ISSN 1744-6848. - ELETTRONICO. - 8:30(2012), pp. 7919-7927. [10.1039/c2sm25532d]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/483963
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