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Evaluation of in vitro efficacy of docetaxel-loaded calcium carbonate aragonite nanoparticles (DTX-CaCO3NP) on 4T1 mouse breast cancer cell line

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Abstract

Cockle shell-derived calcium carbonate nanoparticles have shown promising potentials as slow drug-releasing compounds in cancer chemotherapy. In this study, we evaluated the in vitro efficacy of docetaxel (DTX)-loaded CaCO3NP on 4T1 cell line. This was achieved by evaluating the following: cytotoxicity using MTT assay, fluorescence imaging, apoptosis with Annexin V assay, cell cycle analysis, scanning (SEM) and transmission electron microscopy (TEM), and scratch assay. Based on the results, DTX-CaCO3NP with a DTX concentration of 0.5 μg/mL and above had comparable cytotoxic effects with free DTX at 24 h, while all concentrations had similar cytotoxic effect on 4T1 cells at 48 and 72 h. Fluorescence and apoptosis assay showed a higher (p < 0.05) number of apoptotic cells in both free DTX and DTX-CaCO3NP groups. Cell cycle analysis showed cycle arrest at subG0 and G2/M phases in both treatment groups. SEM showed presence of cellular blebbing, while TEM showed nuclear fragmentation, apoptosis, and vacuolation in the treatment groups. Scratch assay showed lower (p < 0.05) closure in both free DTX and DTX-CaCO3NP groups. The results from this study showed that DTX-CaCO3NP has similar anticancer effects on 4T1 cells as free DTX, and since it has a slow release rate, it is a more preferred substitute for free DTX.

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Acknowledgements

The authors are grateful for funding from Universiti Putra Malaysia Postgraduate Grant Scheme, (GP/IPS/2014/9440300).

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Authors and Affiliations

  1. Department of Veterinary Pre-Clinical Science, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Nahidah Ibrahim Hammadi, Mohd Noor Mohd Hezmee, Intan Shameha Abdul Razak & Zuki Abu Bakar Zakaria

  2. Department of Histology, Faculty of Science, University of Al-Anbar, Al-Anbar, 31005, Iraq

    Nahidah Ibrahim Hammadi

  3. Department of Veterinary Pathology, Faculty of Veterinary Medicine, University of Maiduguri, PMB 1069, Maiduguri, Borno State, 600233, Nigeria

    Yusuf Abba

  4. Department of Pharmacology Faculty of Basic Health Sciences, Bauchi State University Gadau, Gadau, Nigeria

    Aminu Umar Kura

Authors
  1. Nahidah Ibrahim Hammadi
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  2. Yusuf Abba
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  3. Mohd Noor Mohd Hezmee
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  4. Intan Shameha Abdul Razak
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  5. Aminu Umar Kura
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  6. Zuki Abu Bakar Zakaria
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All authors contributed equally to the manuscript and have read and approved the final manuscript.

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Correspondence to Nahidah Ibrahim Hammadi or Zuki Abu Bakar Zakaria.

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All raw data are available on request from Dr. NIH.

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The authors declare that they have no conflict of interest.

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Editor: Tetsuji Okamoto

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Hammadi, N.I., Abba, Y., Hezmee, M.N.M. et al. Evaluation of in vitro efficacy of docetaxel-loaded calcium carbonate aragonite nanoparticles (DTX-CaCO3NP) on 4T1 mouse breast cancer cell line. In Vitro Cell.Dev.Biol.-Animal 53, 896–907 (2017). https://doi.org/10.1007/s11626-017-0197-3

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