Skip to main content
SpringerLink
Log in

Comparison of different methods for preparation of nanochitosan as anticancer agent

  • ORIGINAL PAPER
  • Published:
Polymer Bulletin Aims and scope Submit manuscript

Abstract

Chitosan, the second naturally abundant polysaccharide, has shown promising anticancer activity against many cancer cells. There are various chitosan nanoparticle preparation techniques. This study compared three of these methods, namely, ionotropic gelation, microemulsion, and emulsification solvent diffusion in terms of their product physicochemical and biological properties. To compare different methods, type of chitosan, pH and concentration of chitosan solution were kept constant in all methods. The obtained chitosan nanoparticles were characterized using FTIR, UV–Visible spectroscopy, and SEM. The anticancer activity of the nanoparticles was evaluated by MTT assay in MDA-MB-231 cells at different doses (0.5, 1, 1.5, 2 mg/mL). The morphological alterations of cells were assessed by light inverted microscope. All three methods resulted in nanoparticle formation with the size and zeta potential range of 240–442 nm and + 19.1–34.6 mV, respectively. The ionotropic gelation method yielded smaller nanoparticles with higher zeta potential than those yielded by the microemulsion and emulsification solvent diffusion methods. The cytotoxicity assay showed a dose-dependent effect of nanoparticles. The nanochitosans prepared using the ionotropic gelation, microemulsion, and emulsification solvent diffusion methods showed maximum 77.87%, 63.12%, and 53.17% inhibition against MDA-MB-231 cells, respectively. The results concluded that all obtained nanoparticles have acceptable potency for cytotoxicity against MDA-MB-231 cells with IC50 ranged from 0.89 to 1.67 mg / mL. However, nanoparticles prepared using ionotropic gelation method exhibit the highest anticancer activity. Overall, chitosan nanoparticles obtained using all three methods could serve as anticancer agents and applied in the development of novel antitumor drugs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Data availability

The authors confirm that the data supporting the findings of this study are available within the article.

References

  1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F (2021) Global cancer statistics 2020: globocan estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 71(3):209–249. https://doi.org/10.3322/caac.21660

    Article  PubMed  Google Scholar 

  2. Arem H, Loftfield E (2018) Cancer epidemiology: a survey of modifiable risk factors for prevention and survivorship. Am J Lifestyl Med 12(3):200–210. https://doi.org/10.1177/1559827617700600

    Article  Google Scholar 

  3. Chan H-K, Ismail S (2014) Side effects of chemotherapy among cancer patients in a Malaysian general hospital: experiences, perceptions and informational needs from clinical pharmacists. Asian Pac J Cancer Prev 15(13):5305–5309. https://doi.org/10.7314/apjcp.2014.15.13.5305

    Article  PubMed  Google Scholar 

  4. Ali I, Wani WA, Haque A, Saleem K (2013) Glutamic acid and its derivatives: candidates for rational design of anticancer drugs. Future Med Chem 5(8):961–978. https://doi.org/10.4155/fmc.13.62

    Article  CAS  PubMed  Google Scholar 

  5. Saleem K, Wani WA, Haque A, Lone MN, Hsieh M-F, Jairajpuri MA, Ali I (2013) Synthesis, DNA binding, hemolysis assays and anticancer studies of copper(II), nickel(II) and iron(III) complexes of a pyrazoline-based ligand. Future Med Chem 5(2):135–146. https://doi.org/10.4155/fmc.12.201

    Article  CAS  PubMed  Google Scholar 

  6. Grigore ME (2017) Organic and inorganic nano-systems used in cancer treatment. J Med Res Health Educ https://www.imedpub.com/medical-research-and-health-education/

  7. Dutta S, Ray S (2013) Glutamic acid as anticancer agent: An overview. Saudi Pharm J 21(4):337–343. https://doi.org/10.1016/j.jsps.2012.12.007

    Article  PubMed  PubMed Central  Google Scholar 

  8. Ali I, Wani WA, Saleem K, Wesselinova D (2013) Syntheses, DNA binding and anticancer profiles of L-glutamic acid ligand and c(II) and ruthenium(III) complexes. Med Chem 9:11–21. https://doi.org/10.2174/157340613804488297

    Article  CAS  PubMed  Google Scholar 

  9. Yu Z, Gao L, Chen K, Zhang W, Zhang Q, Li Q, Hu K (2021) Nanoparticles: A new approach to upgrade cancer diagnosis and treatment. Nanoscale Res Lett. https://doi.org/10.1186/s11671-021-03489-z

    Article  PubMed  PubMed Central  Google Scholar 

  10. Joudeh N, Linke D (2022) Nanoparticle classification, physicochemical properties, characterization, and applications: a comprehensive review for biologists. J Nanobiotechnol. https://doi.org/10.1186/s12951-022-01477-8

    Article  Google Scholar 

  11. Vinardell MP, Mitjans M (2015) Antitumor activities of metal oxide nanoparticles. Nanomaterials 5(2):1004–1021. https://doi.org/10.3390/nano5021004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Abarca-Cabrera L, Fraga-García P, Berensmeier S (2021) Bio-nano interactions: binding proteins, polysaccharides, lipids and nucleic acids onto magnetic nanoparticles. Biomater Res 25:1–18. https://doi.org/10.1186/s40824-021-00212-y

    Article  CAS  Google Scholar 

  13. Xu J-J, Zhang W-C, Guo Y-W, Chen X-Y, Zhang Y-N (2022) Metal nanoparticles as a promising technology in targeted cancer treatment. Drug Deliv 29:664–678. https://doi.org/10.1080/10717544.2022.2039804

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Hussain A, Oves M, Alajmi MF, Hussain I, Amir S, Ahmed J, Rehman MT, El-Seed HR, Ali I (2019) Biogenesis of ZnO nanoparticles using Pandanus odorifer leaf extract: anticancer and antimicrobial activities. RSC Adv 9:15357–15369. https://doi.org/10.1039/C9RA01659G

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Bigaj-Józe MJ, Grześkowiak BF (2022) Polymeric nanoparticles wrapped in biological membranes for targeted anticancer treatment. Eur Polym J 176:111427. https://doi.org/10.1016/j.eurpolymj.2022.111427

    Article  CAS  Google Scholar 

  16. Jana S, Sen KK, Gandhi A (2016) Alginate based nanocarriers for drug delivery applications. Curr Pharm Des 22:3399–3410

    Article  CAS  PubMed  Google Scholar 

  17. Masood F (2016) Polymeric nanoparticles for targeted drug delivery system for cancer therapy. Mater Sci Eng C 60:569–578. https://doi.org/10.1016/j.msec.2015.11.067

    Article  CAS  Google Scholar 

  18. Acharya S, Sahoo SK (2011) PLGA nanoparticles containing various anticancer agents and tumour delivery by EPR effect. Adv Drug Deliv Rev 63(3):170–183. https://doi.org/10.1016/j.addr.2010.10.008

    Article  CAS  PubMed  Google Scholar 

  19. Wong KH, Aiping LuA, Chen X, Yang Z (2020) Natural ingredient-based polymeric nanoparticles for cancer treatment. Molecules 25(16):3620. https://doi.org/10.3390/molecules25163620

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ding J, Guo Y (2022) Recent advances in chitosan and its derivatives in cancer treatment. Front Pharmacol l 13:888740. https://doi.org/10.3389/fphar.2022.888740

    Article  CAS  Google Scholar 

  21. Zivarpour P, Hallajzadeh J, Asemi Z, Sadoughi F, Sharifi M (2021) Chitosan as possible inhibitory agents and delivery systems in leukemia. Cancer Cel Int 21(1):544. https://doi.org/10.1186/s12935-021-02243-w

    Article  CAS  Google Scholar 

  22. Key J, Park K (2017) Multicomponent, tumor-homing chitosan nanoparticles for cancer imaging and therapy. Int J Mol Sci 18(3):594. https://doi.org/10.3390/ijms18030594

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Wimardhani YS, Suniarti DF, Freisleben HJ, Wanandi SI, Siregar NC, Ikeda MA (2014) Chitosan exerts anticancer activity through induction of apoptosis and cell cycle arrest in oral cancer cells. J Oral Sci 56:119–126. https://doi.org/10.2334/josnusd.56.119

    Article  CAS  PubMed  Google Scholar 

  24. Divya K, Jisha MS (2018) Chitosan nanoparticles preparation and applications. Environ Chem Lette 16:101–112. https://doi.org/10.1007/s10311-017-0670-y

    Article  CAS  Google Scholar 

  25. Rizeq BR, Younes NN, Rasool K, Nasrallah GK (2019) Synthesis, bioapplications, and toxicity evaluation of chitosan-based nanoparticles. Int J Mol Sci 20(22):5776. https://doi.org/10.3390/ijms20225776

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Babiia O, Wanga Z, Liu G, Martinez EC, Littel-van den Hurk SD, Chen L (2020) Low molecular weight chitosan nanoparticles for CpG oligodeoxynucleotides delivery: impact of molecular weight, degree of deacetylation, and mannosylation on intracellular uptake and cytokine induction. Int J Biol Macromol 159:46–56. https://doi.org/10.1016/j.ijbiomac.2020.05.048

    Article  CAS  Google Scholar 

  27. Antonioua J, Liua F, Majeeda H, Qi J, Yokoyama W, Zhong F (2015) Physicochemical and morphological properties of size-controlled chitosan–tripolyphosphate nanoparticles. Colloids Surf A: Physicochem Eng Asp 465:137–146. https://doi.org/10.1016/j.colsurfa.2014.10.040

    Article  CAS  Google Scholar 

  28. Ngan LTK, Wang S-L, Hiep DM, Luong PM, Vui NT, Dinh TM, Dzung NA (2014) Preparation of chitosan nanoparticles by spray drying, and their antibacterial activity. Res Chem Intermed 40:2165–2175

    Article  CAS  Google Scholar 

  29. Sadat SMA, Jahan ST, Haddadi A (2016) Effects of size and surface charge of polymeric nanoparticles on in vitro and in vivo applications. J Biomater Nanobiotechnol 7:91–108. https://doi.org/10.4236/jbnb.2016.72011

    Article  CAS  Google Scholar 

  30. Hou Z, Zhan C, Jiang Q, Hu Q, Li L, Chang D, Yang X, Wang Y, Li Y, Ye S, Xie L, Yi Y, Zhang Q (2011) Both FA- and mPEG-conjugated chitosan nanoparticles for targeted cellular uptake and enhanced tumor tissue distribution. Nanoscale Res Lett 6(1):563. https://doi.org/10.1186/1556-276X-6-563

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Khanmohammadi M, Elmizadeh H, Ghasemi K (2015) Investigation of size and morphology of chitosan nanoparticles used in drug delivery system employing chemometric technique. Iran J Pharm Res 14(3):665–675. https://doi.org/10.22037/IJPR.2015.1761

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Abdelkader H, Hussain SA, Abdullah N, SuryaniKmaruddin S (2018) Review on micro-encapsulation with chitosan for pharmaceuticals applications. MOJ Curr Res Rev 1(2):77–84. https://doi.org/10.15406/mojcrr.2018.01.00013

    Article  Google Scholar 

  33. Fan W, Yan W, Xu Z, Ni H (2012) Formation mechanism of monodisperse, low molecular weight chitosan nanoparticles by ionic gelation technique. Colloids Surf B Biointerfaces 90:21–27. https://doi.org/10.1016/j.colsurfb.2011.09.042

    Article  CAS  PubMed  Google Scholar 

  34. Hasanzadeh Kafshgari M, Khorram M, Mansouri M, Samimi A, Osfouri S (2012) Preparation of alginate and chitosan nanoparticles using a new reverse micellar system. Iran Polym J 21(2):99–107. https://doi.org/10.1007/s13726-011-0010-1

    Article  CAS  Google Scholar 

  35. Grenha A (2012) Chitosan nanoparticles: A survey of preparation methods. J Drug Target 20(4):291–300. https://doi.org/10.3109/1061186X.2011.654121

    Article  CAS  PubMed  Google Scholar 

  36. Rebbouh-Nouiouat F, Marie France ME, Laraba-Djebari F (2020) Chitosan nanoparticles as a delivery platform for neurotoxin II from Androctonus australis hector scorpion venom: Assessment of toxicity and immunogenicity. Acta Trop 205(4):105353. https://doi.org/10.1016/j.actatropica.2020.105353

    Article  CAS  Google Scholar 

  37. Thamilarasan V, Sethuraman V, Gopinath K, Balalakshmi C, Govindarajan M, Mothana RAA, Siddiqui NA, Khaled JM, Benelli K (2018) Single step fabrication of chitosan nanocrystals using penaeus semisulcatus: potential as new insecticides, antimicrobials and plant growth. J Clust Sci 29:375–384. https://doi.org/10.1007/s10876-018-1342-1

    Article  CAS  Google Scholar 

  38. Song C, Yu H, Zhang M, Yang Y, Zhang G (2013) Physicochemical properties and antioxidant activity of chitosan from the blowfly Chrysomya megacephala larvae. Int J Biol Macromol 60:347–354. https://doi.org/10.1016/j.ijbiomac.2013.05.039

    Article  CAS  PubMed  Google Scholar 

  39. Huang K, Ma H, Liu J, Huo S, Kumar A, Wei T, Zhang X, Jin S, Gan Y, Wang PC, He S, Zhang X, Liang X-J (2012) Size-dependent localization and penetration of ultrasmall gold nanoparticles in cancer cells, multicellular spheroids, and tumors in vivo. ACS Nano 6:4483–4493. https://doi.org/10.1021/nn301282m

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Ko W-K, Heo DN, Moon H-J, Lee SJ, Bae MS, Lee JB, Sun I-C, Jeon HB, Park HK, Kwon IK (2015) The effect of gold nanoparticle size on osteogenic differentiation of adipose-derived stem cells. J Colloid Interface Sci 438:68–76. https://doi.org/10.1016/j.jcis.2014.08.058

    Article  CAS  PubMed  Google Scholar 

  41. Honary S, Zahir F (2013) Effect of zeta potential on the properties of nano-drug delivery systems. Trop J Pharm Res 12:265–273. https://doi.org/10.4314/tjpr.v12i2.20

    Article  CAS  Google Scholar 

  42. Jeon S, Clavadetscher J, Lee DK, Chankeshwara SV, Bradley M, Cho WS (2018) Surface charge-dependent cellular up-take of polystyrene nanoparticles. Nanomaterials 8:1028. https://doi.org/10.3390/nano8121028

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Li S, Malmstadt N (2013) Deformation and poration of lipid bilayer membranes by cationic nanoparticles. Soft Matter 9(20):4969–4976. https://doi.org/10.1039/C3SM27578G

    Article  CAS  Google Scholar 

  44. SohrabiKashani A, Packirisamy M (2021) Cancer-nano-interaction: from cellular uptake to mechanobiological responses. Int J Mol Sci 22:9587. https://doi.org/10.3390/ijms22179587

    Article  CAS  Google Scholar 

  45. Mizushima N (2018) A brief history of autophagy from cell biology to physiology and disease. Nat Cell Biol 20:521–527. https://doi.org/10.1038/s41556-018-0092-5

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Iranian Research Organization for Science and Technology [grant number 4404]

Author information

Authors and Affiliations

  1. Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran

    Forouh sadat Hassani, Mahnaz Hadizadeh & Davood Zare

  2. Amirkabir Nanotechnology Research Institute (ANTRI), Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran

    Saeedeh Mazinani

Authors
  1. Forouh sadat Hassani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mahnaz Hadizadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Davood Zare
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Saeedeh Mazinani
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by FsH and MH. The first draft of the manuscript was written by FsH and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Mahnaz Hadizadeh.

Ethics declarations

Conflict of interest

All the authors declare that there is no conflict of interest.

Ethical approval

We the undersigned declare that this manuscript is original, has not been published before and is not currently being considered for publication elsewhere. We confirm that the manuscript has been read and approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed. We further confirm that the order of authors listed in the manuscript has been approved by all of us. We understand that the corresponding author is the sole contact for the editorial process. She is responsible for communicating with the other authors about progress, submissions of revisions and final approval of proofs signed by all authors as follows: Forouh Sadat Hassani, Mahnaz Hadizadeh, Davood Zare, and Saeedeh Mazinani.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hassani, F.s., Hadizadeh, M., Zare, D. et al. Comparison of different methods for preparation of nanochitosan as anticancer agent. Polym. Bull. 81, 827–842 (2024). https://doi.org/10.1007/s00289-023-04739-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00289-023-04739-z

Keywords

Search

Navigation