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Chitosan-Coated Lipid-Core Nanocapsules Functionalized with Gold-III and Bevacizumab Induced In Vitro Cytotoxicity against C6 Cell Line and In Vivo Potent Antiangiogenic Activity

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Abstract

Purpose

Bevacizumab (BCZ) is a recombinant monoclonal antibody that inhibits the biological activity of the vascular endothelial growth factor, which has an important role in angiogenesis for tumoral growth and progression. In this way, our objective was to develop chitosan-coated lipid-core nanocapsules functionalized with BCZ by an organometallic complex using gold-III.

Methods

The formulation was produced and characterized in relation to physicochemical characteristics. Furthermore, the antitumoral and antiangiogenic activities were evaluated against C6 glioma cell line and chicken embryo chorioallantoic membrane (CAM), respectively.

Results

Final formulation showed nanometric size, narrow polydispersity, positive zeta potential and gold clusters size lower than 2 nm. BCZ in aqueous solution (0.01–0.10 μmol L−1) did not show cytotoxic activity in vitro against C6 glioma cell line; although, MLNC-Au-BCZ showed cytotoxicity with a median inhibition concentration of 30 nmol L−1 of BCZ. Moreover, MLNC-Au-BCZ demonstrated cellular internalization dependent on incubation time and BCZ concentration. BCZ solution did not induce significant apoptosis as compared to MLNC-Au-BCZ within 24 h of treatment. CAM assay evidenced potent antiangiogenic activity for MLNC-Au-BCZ, representing a decrease of 5.6 times in BCZ dose comparing to BCZ solution.

Conclusion

MLNC-Au-BCZ is a promising product for the treatment of solid tumors.

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Abbreviations

BCZ:

Bevacizumab

CAM:

Chorioallantoic membrane

CCT:

Caprylic/capric triglyceride

DLS:

Dynamic light scattering

NTA:

Nanoparticle tracking analysis

DMEM:

Dulbecco’s Modified Eagle’s Medium

FTIR:

Fourier transform infrared spectroscopy

LNC:

Lipid-core nanocapsules

LNC-Au:

Lipid-core nanocapsules functionalized with gold-III

LNC-Au-Phe:

Lipid-core nanocapsules functionalized with gold-III and phenylalanine

MLNC:

Chitosan-coated lipid-core nanocapsules

MLNC-Au:

Chitosan-coated lipid-core nanocapsules functionalized with gold-III

MLNC-Au-BCZ:

Chitosan-coated lipid-core nanocapsules functionalized with gold-III and bevacizumab

MLNC-Au-Phe:

Chitosan-coated lipid-core nanocapsules functionalized with gold-III and phenylalanine

MLNCf-Au-BCZ:

Fluorescent chitosan-coated lipid-core nanocapsules functionalized with gold-III and bevacizumab

P80:

Polysorbate 80

PCL:

Poly(ε-caprolactone)

PDI:

Polydispersity index

SL:

Soybean lecithin

SM:

Sorbitan monostearate

TEM:

Transmission electron microscopy

VEGF:

Vascular endothelial growth factor

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Acknowledgements

The authors thank Professor Maria Palmira Daflon Gremião (Universidade Estadual Paulista em Araraquara, UNESP) for the Avastin® sample; Professor Sergio Luiz Vieira and Thiago Luiz Noetzold (Aviário de Ensino e Pesquisa do Departamento de Zootecnia, Universidade Federal do Rio Grande do Sul, UFRGS) for the eggs supply; and the Brazilian Agencies (CAPES, CNPq and FAPERGS). This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES, finance code 001), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS: 17/2551-0001002-7; PRONEX FAPERGS-CNPq 16/2551-0000467-6) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (Research grant 305301/2014-4 and INCT-Nanofarma grant 465687/2014-8).

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Authors and Affiliations

  1. Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga, 2752, Porto Alegre, RS, 90610-000, Brazil

    Aline de Cristo Soares Alves, Danieli Rosane Dallemole, Rodrigo Cé, Silvia Stanisçuaski Guterres & Adriana Raffin Pohlmann

  2. Departamento de Química Inorgânica, Instituto de Química, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500, Porto Alegre, RS, 91501-970, Brazil

    Vladimir Lavayen

  3. Departamento de Bioquímica, Instituto de Ciências da Saúde, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600, Porto Alegre, RS, 90035-003, Brazil

    Fabrício Figueiró & Ana Maria Oliveira Battastini

  4. Programa de Pós-Graduação em Ciências Biológicas: Bioquímica, Instituto de Ciências da Saúde, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600, Porto Alegre, RS, 90035-003, Brazil

    Fabrício Figueiró & Amanda de Fraga Dias

  5. Departamento de Química Orgânica, Instituto de Química, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500, Porto Alegre, RS, 91501-970, Brazil

    Adriana Raffin Pohlmann

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  1. Aline de Cristo Soares Alves
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  2. Vladimir Lavayen
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  3. Fabrício Figueiró
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de Cristo Soares Alves, A., Lavayen, V., Figueiró, F. et al. Chitosan-Coated Lipid-Core Nanocapsules Functionalized with Gold-III and Bevacizumab Induced In Vitro Cytotoxicity against C6 Cell Line and In Vivo Potent Antiangiogenic Activity. Pharm Res 37, 91 (2020). https://doi.org/10.1007/s11095-020-02804-0

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