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Cell J. 2019 Apr;21(1):14-26. doi: 10.22074/cellj.2019.5691. Epub 2018 Nov 18.

In Vitro Cytotoxicity of Folate-Silica-Gold Nanorods on Mouse Acute Lymphoblastic Leukemia and Spermatogonial Cells.

Author information

1
Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
2
Department of Medical Physics, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
3
Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran.
4
Department of Medical Nanotechnology, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
5
Neuroscience Research Center, Iran University of Medical Sciences, Tehran, Iran.
6
Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
7
Clinical Nanomedicine Laboratory, ENT-Head and Neck Research Center, Hazrat Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran.
8
Oncopathology Research Center and Dep Pathology, Faculty of Medicine Iran University of Medical Sciences, Tehran, Iran.
9
VetCell Therapeutics, Daimler St, Santa Ana CA, USA.
10
Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran. Electronic Address: koruji.m@iums.ac.ir.

Abstract

Objective:

The purpose of this study was to evaluate in vitro cytotoxicity of gold nanorods (GNRs) on the viability of spermatogonial cells (SSCs) and mouse acute lymphoblastic leukemia cells (EL4s).

Materials and Methods:

In this experimental study, SSCs were isolated from the neonate mice, following enzymatic digestion and differential plating. GNRs were synthesized, then modified by silica and finally conjugated with folic acid to form F-Si-GNRs. Different doses of F-Si-GNRs (25, 50, 75, 100, 125 and 140 μM) were used on SSCs and EL4s. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) proliferation assay was performed to examine the GNRs toxicity. Flow cytometry was used to confirm the identity of the EL4s and SSCs. Also, the identity and functionality of SSCs were determined by the expression of specific spermatogonial genes and transplantation into recipient testes. Apoptosis was determined by flow cytometry using an annexin V/propidium iodide (PI) kit.

Results:

Flow cytometry showed that SSCs and EL4s were positive for Plzf and H-2kb, respectively. The viability percentage of SSCs and EL4s that were treated with 25, 50, 75, 100, 125 and 140 μM of F-Si-GNRs was 65.33 ± 3.51%, 60 ± 3.6%, 51.33 ± 3.51%, 49 ± 3%, 30.66 ± 2.08% and 16.33 ± 2.51% for SSCs and 57.66 ± 0.57%, 54.66 ± 1.5%, 39.66 ± 1.52%, 12.33 ± 2.51%, 10 ± 1% and 5.66 ± 1.15% for EL4s respectively. The results of the MTT assay indicated that 100 μM is the optimal dose to reach the highest and lowest level of cell death in EL4s and in SSCs, respectively.

Conclusion:

Cell death increased with increasing concentrations of F-Si-GNRs. Following utilization of F-Si-GNRs, there was a significant difference in the extent of apoptosis between cancer cells and SSCs.

KEYWORDS:

Acute Lymphoblastic Leukemia Cells; Cytotoxicity; Folic Acid; Gold Nanorods; Spermatogonial Cells

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