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Labeling PLA-PEG nanocarriers with IR780: physical entrapment versus covalent attachment to polylactide

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Abstract

Near-infrared fluorescent dyes, such as IR780, are promising theranostics, acting as photosensitizers for photodynamic therapy and in vivo tracers in image-guided diagnosis. This work compared the uptake by macrophage-like cells of IR780 either physically associated or covalently attached to poly(D,L-lactide) (PLA) formulated as polymeric nanocapsules (NC) from a blend of PLA homopolymer and PLA-PEG block copolymer. The physicochemical characterization of both NC was conducted using asymmetric flow field-flow fractionation (AF4) analysis with static and dynamic light scattering and atomic force microscopy. The interaction of IR780 with serum proteins was evidenced by AF4 with fluorescence detection and flow cytometry in cell uptake studies. The average diameters of NC were around 120 nm and zeta potentials close to -40 mV for all NC. NC uptake by cells in different media and experimental conditions shows significantly lower fluorescence intensities for IR780 covalently linked to PLA and correspondingly low quantitative uptake. Different mechanisms of internalization were evidenced depending on the IR780 type of association to NC. Serum proteins mediate IR780 interaction with cells in a dose-dependent manner. Our results show that non-covalently linked IR780 was released from NC and accumulated in macrophage cells. Oppositely, IR780 conjugated to PLA provides stable association with NC, and its fluorescence is representative of cell uptake of the nanocarrier itself. This work strongly reinforces the importance of covalent attachment of a fluorescence dye such as IR780 to the nanocarrier to study their interaction with cells in vitro and to obtain reliable tracking in image-guided therapy.

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Acknowledgments

The authors thank CAPES/Brazil and FAPEMIG scholarships for postgraduation students. We acknowledge Dra. Nívia C.N. Paiva for her skillful assistance with confocal laser microscopy and Prof. F. G. Araújo for the access to the AFM equipment at the NanoLab/REDEMAT/UFOP acquired with financial support of CT-Infra FINEP/Brazil.

Funding

We received financial support from Brazilian funding agencies: INCT-NANOFARMA (Grant #2014/50928-2) and CNPq-grants (# BJT 400889/2014-5, #310463/2015-7 and #481872/2013-2) FAPEMIG-grants (#APQ-02864-16, APQ-02576-18 and NANOBIOMG Network-RED-00007-14) and PROPP/UFOP (grant to APMB).

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Authors and Affiliations

  1. Laboratory of Pharmaceutics and Nanotechnology, Pharmacy School, Federal University of Ouro Preto, Minas Gerais, Brazil

    Marina Guimarães Carvalho Machado, Gwenaelle Pound-Lana, Maria Alice de Oliveira, Elisa Gomes Lanna, Márcia Célia Pacheco Fialho & Vanessa Carla Furtado Mosqueira

  2. Laboratory of Microscopy, Department of Physics, Exact and Biological Sciences Institute, Federal University of Ouro Preto, Minas Gerais, Brazil

    Ana Carolina Ferreira de Brito & Ana Paula Moreira Barboza

  3. Department of Clinical Analysis, Pharmacy School and Nucleus of Research in Biological Sciences (NUPEB), Federal University of Ouro Preto, Minas Gerais, Brazil

    Rodrigo Dian de Oliveira Aguiar-Soares

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  1. Marina Guimarães Carvalho Machado
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  2. Gwenaelle Pound-Lana
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  3. Maria Alice de Oliveira
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  4. Elisa Gomes Lanna
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Contributions

Conceptualization [MGCM, GPL, VCFM]; methodology [MGCM, GPL, MAO, EGL, MCPF, ACFB, APMB, RDOAS]; formal analysis and investigation [MGCM, GPL, VCFM]; original draft preparation [MGCM]; writing, review and editing [MGCM, GPL, VCFM]; funding acquisition [VCFM]; resources [VCFM]; supervision [GPL, VCFM].

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Correspondence to Vanessa Carla Furtado Mosqueira.

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Machado, M.G.C., Pound-Lana, G., de Oliveira, M.A. et al. Labeling PLA-PEG nanocarriers with IR780: physical entrapment versus covalent attachment to polylactide. Drug Deliv. and Transl. Res. 10, 1626–1643 (2020). https://doi.org/10.1007/s13346-020-00812-6

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