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Cell uptake mechanisms of PAMAM G4-FITC dendrimer in human myometrial cells

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Abstract

The high incidence and severity of diseases which involve smooth muscle dysfunction dictates the need of continued search for novel therapeutic strategies to treat these conditions. Dendrimers are branched macromolecules with multiple end-groups that can be functionalized for applications which include drug delivery. There is no data regarding the cellular uptake mechanisms used by dendrimers in smooth muscle human myometrial cells (HMC). Polyamidoamine G4 dendrimers were conjugated with fluorescein isothiocyanate (FITC) and the resulting conjugate (G4-FITC) was characterized using high-performance liquid chromatography, nuclear magnetic resonance, and atomic force microscopy. G4-FITC showed to have no significant effect on the primary culture HMC viability up to 48 h. HMC incubated with G4-FITC were analyzed by laser confocal microscopy. Peri-nuclear fluorescence distribution was observed at 5 h of incubation or more (24, 36, and 48 h). At 24 h, G4-FITC partially co-localized with lysotracker. Uptake of G4-FITC by HMC was slightly inhibited by filipin (8.0 ± 3.9 %) and significantly inhibited by chlorpromazine (63.5 ± 3.7 %). In non-electroporated HMC, G4-FITC was never observed inside the cell nucleus. Interestingly, we detected G4-FITC inside the nuclear domain of some electroporated cells. Thus, electroporation changed intracellular G4-FITC localization. Isolated nuclei of HMC incubated with G4-FITC showed fluorescence signal inside the nuclear domain. The results suggest that in HMC, G4-FITC is taken up by clathrin-mediated endocytosis with endosomal and lysosomal localization at 24 h. The combination of electroporation and dendrimers could be an interesting technology to electrotransfer drugs into smooth muscle cells cytosol and nuclei.

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Acknowledgments

The research described in this article was supported by the Scientific Research in Cancer Support Program of “Comisión Honoraria de Lucha contra el Cáncer,” by the “Agencia Nacional de Investigación e Innovación” from Uruguay (ANII, FCE_2009_1_2887) and by PEDECIBA. We would like to thank Pereira Rossell Hospital, Montevideo, Uruguay, for providing myometrial tissue samples. We deeply appreciate the collaboration of: Andrés Alberro, from our laboratory, in preparing the figures and Jelver Sierra, from “Grupo de estudo de Interações entre Macro e Micromoléculas”—Universidade Federal de Santa Catarina—in making zetasizer measurements. English linguistic precision was revised by Ms. Rita Pessano.

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Authors and Affiliations

  1. Laboratorio de Señalización Celular y Nanobiología, Instituto de Investigaciones Biológicas Clemente Estable, Av. Italia 3318, 11600, Montevideo, Uruguay

    Natalia Oddone, Ana I. Zambrana, Marcos Tassano & Juan C. Benech

  2. Laboratorio de Radiofarmacia, Centro de Investigaciones Nucleares, Facultad de Ciencias, Universidad de la República, Mataojo 2055, 11400, Montevideo, Uruguay

    Marcos Tassano & Pablo Cabral

  3. Grupo de Química Medicinal, Instituto de Química Biológica, Facultad de Ciencias-Facultad de Química, Universidad de la República, Iguá 4225 Esq. Mataojo, 11400, Montevideo, Uruguay

    Williams Porcal

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  1. Natalia Oddone
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  2. Ana I. Zambrana
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  3. Marcos Tassano
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  5. Pablo Cabral
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  6. Juan C. Benech
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Correspondence to Juan C. Benech.

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Oddone, N., Zambrana, A.I., Tassano, M. et al. Cell uptake mechanisms of PAMAM G4-FITC dendrimer in human myometrial cells. J Nanopart Res 15, 1776 (2013). https://doi.org/10.1007/s11051-013-1776-1

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  • DOI: https://doi.org/10.1007/s11051-013-1776-1

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