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PLA-PEG nanospheres decorated with phage display selected peptides as biomarkers for detection of human colorectal adenocarcinoma

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Abstract

Peptide-mediated targeting to colorectal cancer can increase selectivity and specificity of this cancer diagnosis acting as biomarkers. The present work aimed to select peptides using the phage display technique and associate the peptides with polymeric nanospheres in order to evaluate their cytotoxicity and selectivity during cell interaction with Caco-2 human colon tumor cell line. Two peptides identified by phage display (peptide-1 and peptide-2) were synthesized and exhibited purity higher than 84%. Poly(lactic acid)-block-polyethylene glycol nanospheres were prepared by nanoprecipitation and double emulsion methods in order to load the two peptides. Nanoparticles ranged in size from 114 to 150 nm and peptide encapsulation efficiency varied from 16 to 32%, depending on the methodology. No cytotoxic activity was observed towards Caco-2 tumor cell line, either free or loaded peptides in concentrations up to 3 μM at incubation times of 6 and 24 h, indicating safety as biomarkers. Fluorescein isothiocyanate–labeled peptides allowed evaluating selective interactions with Caco-2 cells, where peptide-1 entrapped in nanospheres showed greater intensity of co-localized cell fluorescence, in comparison to peptide-2. Peptide-1 loaded in nanospheres revealed promising to be investigated in further studies of selectivity with other human colon rectal cells as a potential biomarker.

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Acknowledgments

This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001. We also thank the financial support from the following Brazilian funding agencies: CNPq grants (#310463/2015-7 and #481872/2013-2) and FAPEMIG grants (#APQ-02194-16, #APQ-02864-16, APQ-02576-18, and NANOBIOMG Network-RED-00007-14). VCFM is a productivity research fellow of CNPq. We acknowledge Raoni P. Siqueira for skillful help with cell culture and Prof. F.G. Araújo for giving access to AFM equipment in NanoLab/REDEMAT/UFOP acquired with financial support of CT-Infra FINEP/Brazil.

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Authors and Affiliations

  1. Laboratory of Pharmaceutics and Nanobiotechnology (LDGNano), School of Pharmacy, Universidade Federal de Ouro Preto, Minas Gerais, Brazil

    Aline de Souza, Elisa Gomes Lanna, Vanessa Carla Furtado Mosqueira & Karina Taciana Santos Rubio

  2. Laboratory of Enzymology and Proteomics, NUPEB, Universidade Federal de Ouro Preto, Minas Gerais, Brazil

    William Castro-Borges & Milton Hércules Guerra de Andrade

  3. Laboratory of Nanobiotechnology, Institute of Biotechnology, Universidade Federal de Uberlandia, Minas Gerais, Brazil

    Yara Cristina de Paiva Maia & Luiz Ricardo Goulart

  4. Department of Physics, Exact and Biological Sciences Institute, Universidade Federal de Ouro Preto, Minas Gerais, Brazil

    Ana Carolina Ferreira de Brito & Ana Paula Moreira Barboza

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  1. Aline de Souza
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  2. William Castro-Borges
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Contributions

Conceptualization: AS, VCFM, WCB, and KTSR. Methodology: AS, WCB, MHGA, YCPM, LRGF, EGL, ACFB, APMB, VCFM, and KTSR. Formal analysis and investigation: AS, WCB, APMB, VCFM, and KTSR. Original draft preparation: AS and KTSR. Writing—review and editing: AS, WCB, VCFM, and KTSR. Funding acquisition: KTSR, WCB, and VCFM. Resources: KTSR and VCFM. Supervision: VCFM and KTSR.

Corresponding author

Correspondence to Vanessa Carla Furtado Mosqueira.

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Conflict of interest

A. Souza, W. Castro-Borges, M.H.G. Andrade, Y.C.P. Maia, L.R.G. Filho, E.G. Lanna, A.C.F. Brito, A.P.M. Barboza, V.C.F. Mosqueira, and K.T.S. Rubio declare that they have no conflict of interest.

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de Souza, A., Castro-Borges, W., de Andrade, M.H.G. et al. PLA-PEG nanospheres decorated with phage display selected peptides as biomarkers for detection of human colorectal adenocarcinoma. Drug Deliv. and Transl. Res. 10, 1771–1787 (2020). https://doi.org/10.1007/s13346-020-00826-0

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