Synthesis, Characterization and Cytotoxicity Investigation of Chitosan-Amino Acid Derivatives Nanoparticles in Human Breast Cancer Cell Lines

Cytotoxicity Investigation of Chitosan-Amino Acid Derivatives Nanoparticles

  • Maysoon Zaboon University of Basrah-Iraq
  • Afrodet Saleh University of Basrah-Iraq
  • Hadi Al-Lami University of Basrah-Iraq
Keywords: Chitosan-grafted-amino acids, nanoparticles, breast cancer cell lines, cytotoxicity, MTT


Abstract. The presence of reactive primary amines in the backbone structure of chitosan enables the derivatization with different functional groups and thereby improving and expanding its properties, such as solubility and mucoadhesiveness, for biomedical applications. In this work, chitosan was grafted with different sources of amino acids (Histidine, Aspartic acid, Glutamic acid, Glycine-Aspartic acid, and Glycine-Glutamic acid), Chitosan and its grafted amino acid derivatives were obtained in very good yield, and they were characterized by Fourier-Transform Infrared Spectroscopy (FTIR), and the resulted spectra confirmed the right structures of chitosan and its different synthesized derivatives. The chitosan and its amino acid derivatives were converted to nanoparticles in size by subjecting them to the sonication method. The Scanning Electron Microscope (SEM) was used to determine the shape and size of the prepared polymeric nanoparticles and the average nanoparticle size counted by the Image-J program. The micrographs revealed that the nanoparticles with spherical shapes and with different sizes were gained, but in general, they are less than 100nm in diameters. In vitro cytotoxicity of chitosan and chitosan derivatives prepared NPs were determined as MTT assay, against different three types of human breast cancer cell lines which are BT cell lines, MCF-7 cell lines, and SKBR3 cell lines. The cell proliferation of each type of breast cancer cell line has appeared to a highly significant decrease (p<0.001), with all types of tested NPs polymers in comparison with the positive control samples, through different periods of the experiment (24, 48, and 72 hours).


Resumen. La presencia de aminas primarias reactivas en la estructura del quitosano permite su funcionalización con diferentes grupos funcionales, mejorando y expandiendo sus propiedades, por ejemplo, solubilidad y mucoadhesividad, para aplicaciones biomédicas. En este trabajo se injertó quitosano con diferentes fuentes de aminoácidos (histidina, ácido aspártico, ácido glutámico, glicina-ácido aspártico y glicina-ácido glutámico). Los derivados de quitosano injertados con aminoácidos se obtuvieron con muy buen rendimiento. La caracterización por espectroscopía infrarroja (FTIR) confirmó la funcionalización del quitosano. Después de sonicación y una caracterización por microscopía electrónoica de barrido (SEM), se confirmó que el tamaño del quitosano y sus derivados con aminoácidos pueden clasificarse como nanopartículas. Las micrografías revelaron que las nanopartículas tienen formas esféricas y son de diferentes tamaños, pero en general, son menores a 100 nm de diámetro. La citotoxicidad in vitro de las nanopartículas de quitosano y derivados de quitosano se determinó como ensayos MTT frente a tres tipos diferentes de líneas celulares de cáncer de mama humano, a saber, líneas celulares BT, líneas celulares MCF-7 y líneas celulares SKBR3. La proliferación celular de cada tipo de línea celular de cáncer de mama mostró una disminución significativa (p <0.001), con todos los tipos de polímeros NP probados en comparación con las muestras de control positivo, a lo largo de diferentes períodos del experimento (24, 48, y 72 horas).

Author Biographies

Maysoon Zaboon, University of Basrah-Iraq

Department of Chemistry, College of Science

Afrodet Saleh, University of Basrah-Iraq

Department of Pathological Analysis, College of Science

Hadi Al-Lami, University of Basrah-Iraq

Department of Chemistry, College of Science


Song, Z.; Li G.; Guan, F.; Liu, W. Polymers 2018, 10, 389-402. DOI:

Ibitoye, E. B.; Lokman, I. H.; Hezmee, M. N. M.; Goh, Y. M.; Zuki, A. B. Z.; Jimoh, A. A. Biomed. Mater. 2018, 13, 1-13. DOI:

Mutasher, S. H.; Salih, A. A.; Al-Lami, H. S. Der. Pharma. Chemica 2016, 8, 125-134.

Li, Z.; Yang, F.; Yang, R. Int. J. Biol. Macromol. 2015, 75, 378-387. DOI:

Guţoaia, A. Chitosan Nanoplexes for target siRNA deliver, Ph.D. thesis, the University of Tübingen, aus Galaţi, Rumänien, 2015.

Raftery, R; O’Brien, R. F. J.; Cryan, S. Molecules 2013, 18, 5611- 5647. DOI: https://pubmed/ 10.3390/molecules18055611

Jennings, J. A.; Bumgardner, J. D. Chitosan Based Biomaterials, Volume 2: Tissue Engineering and Therapeutics, 1st ed.; Elsevier Ltd.: United Kingdom, 2017, 1-10.

Carballal, B.; Fernández, E. F.; Goycoolea, F. M. Polymers (Basel) 2018, 10, 444, 1-51. DOI: https://ncbi/10.3390/polym10040444

Zhang, P.; Wagner, E., Top Curr. Chem. (Z) 2017, 375, 1-39. DOI: https://pubmed.ncbi/ 10.1007/s41061-017-0112-0

Gad, A.; Kydd, J.; Piel, B.; Rai1, P. Int. J. Nanomed. Nanosurg. 2016, 2, 1-12. DOI: https://ncbi/10.16966/2470-3206.116

Prabhu, R. H.; Patravale, V. B.; Joshi, M. D. Int. J. Nanomed. 2015, 10, 1001-1018. DOI: https://ncbi/10.2147/IJN.S56932

Chen, G.; Gong, R. Saudi Pharm. J. 2016, 24, 250-253. DOI: https://ncbi/10.1016/j.jsps.2016.04.008

Lounis, F.M.; Chamieh, J.; Leclercq, L.; Gonzalez, P.; Rossi, J.; Cottet, H. Polymers 2018, 10, 45, 1-16. DOI: https://mdpi/10.3390/polym10010045

Hong, C. A.; Sonm, H. Y.; Nam, Y.S. Nature Sci. Rep. 2018, 8, 7738, 1-7. DOI: https://nature/10.1038/s41598-018-25655-7

Perni, S.; Prokopovich, P. Nanom.: Nanotech. Bio. Med. 2017, 13, 539-548. DOI:

Gabano, E.; Quental, L.; Perin, E.; Silva, F.; Raposinho, P.; Paulo, A.; Ravera, M. Inorganics 2018, 6, 1-13. DOI:

Alshraim, M. O.; Sangi, S.; Harisa, G. I.; Alomrani, A. H.; Yusuf O.; Badran, M. M. Molecules 2019, 24, 1-14.DOI: https://pubmed.ncbi/10.3390/molecules24020250

Hajipour, A. R.; Hosseinia, S. M.; Jajarmi, S., New J. Chem. 2017, 41, 7447-7452. DOI:

Galhoum, A. A.; Hassanc, K. M.; Desouky, O. A.; Masoud, A.; Akashid, M. T.; Sakaid Y.; Guibalb, E. React. Funct. Polym. 2017, 113, 13-22. DOI:

Rani, M.; Agarwal, A.; Negi, Y. S. J. Biomat. Nanobiotech. 2011, 2, 71-84. DOI:

Rani, M.; Agarwal, A.; Maharana, T; Negi, Y. S. Afr. J. Pharm. Pharmacol. 2010, 4, 35-54. URL:

Silva, V. J. D. Preparation and characterization of chitosan nanoparticles for gene delivery, M.Sc. thesis, University of Lisbon, Portugal, November 2013, 31-32.

Mosmann, T. J Immunol Methods 1983, 65, 55-63. DOI:

El-Ghaffar, M. A. A.; Akl, M. A.; Kamel, A .M.; Hashem, M. S., Egypt J. Chem. 2017, 60, 507-518. DOI:

Zaboon, M. H. Synthesis and Characterization of some New Polymeric Chitosan Nanoparticles Derivatives as Antitumor for Human Breast Cancer Cell Lines, Ph.D. thesis, University of Basrah, Iraq, Janaury 2019, 44-63.

Ferrari, M.; Fornasiero, M. C.; Isetta, A. M. J. Immun. Meth. 1990, 131, 165-172. DOI: https://pubmed.ncbi.10.1016/0022-1759(90)90187-z

Casettari, L.; Vllasaliu, D.; Lam, J. K.; Soliman, M; Illum, L. Biomaterials 2012, 33, 7565–7583. DOI:

Al-Lami, H. S.; Zaboon, M. H.; Saleh, A. A. IOP Conf. Ser.: Mater. Sci. Eng. 2020, 871, 1-15. DOI:

Wernersson, E.; Heyda, J.; Kubícková, A.; Krízek, T.; Coufal, P.; Jungwirth, P. J. Phys. Chem. B. 2010, 114:11934–11941. DOI:

Raja, M. A.; Zeenat, S. Arif, M.; Liu, C. Int. J. Nanomed. 2016, 11, 4397–4412.DOI

Layek, B.; Singh, J. Biomacromolecules 2013, 14, 485-494. DOI: