Current Issue

Volume 21, Number 1, Apr-Jun(Spring) 2019 , Serial Number: 81 Pages: 14-26

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


Neda Eslahi, Ph.D, 1, Ali Shakeri-Zadeh, Ph.D, 2, Khadijeh Ashtari, Ph.D, 3, 4, Vahid Pirhajati-Mahabadi, Ph.D, 5, Tahereh Tohidi Moghadam, Ph.D., 6, Ronak Shabani, Ph.D., 1, Kamran Kamrava, M.D., 7, Zahra Madjd, Ph.D., 8, Chad Maki, D.V.M., 9, Hamid Reza Asgari, Ph.D., 1, Morteza Koruji, Ph.D., 1, 3, *,
Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
Department of Medical Physics, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran
Department of Medical Nanotechnology, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
Neuroscience Research Center, Iran University of Medical Sciences, Tehran, Iran
Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
Clinical Nanomedicine Laboratory, ENT-Head and Neck Research Center, Hazrat Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran
Oncopathology Research Center and Dep Pathology, Faculty of Medicine Iran University of Medical Sciences, Tehran, Iran
VetCell Therapeutics, Daimler St, Santa Ana CA, USA
*Corresponding Addresses: P.O.Box: 354-14665 Cellular and Molecular Research Center Iran University of Medical Sciences Tehran Iran Department of Anatomical Sciences School of Medicine Iran University of Medical Sciences Tehran Iran Email: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.