In vitro and in vivo studies of cytotoxic effects of FeSO4 nanoparticles

Elahe Moeini,Hadi Ebrahimnejad,Nahid Askari,Mojtaba Bonyadian,Iraj Karimi

doi:10.34172/jsums.2021.01

Abstract

Background and aims: Using iron as a food additive usually causes undesirable sensory changes and side effects in humans. In this study, we made iron (Fe) nanoparticles (NPs) and studied the cytotoxicity of FeSO4 bulk and NPs on HT-29 cells and different doses of these particles on rat intestine. Methods: Particle size of nanoscale was achieved by mechanical technique. Iron particles were characterized using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The effect of iron particles with different concentrations (6.25, 3.125, and 1.57 mM/mL) on the colon cell line was performed using the MTT assay at 24, 48, and 72 hours. Apoptosis and necrosis of the cells were assessed using Annexin V-FITC staining and propidium iodide (PI) at 24 h. In an in vivo study, Taftoon bread was produced from fortified wheat flour with FeSO4 bulk and NPs, which are recommended in human diet (9, 18, and 27 mg of elemental iron/kg flour). Wistar rats were fed daily with fortified bread for 21 days and their colon and small intestine were then evaluated histopathologically. Statistical analyses were performed using SPSS 22.0 software by chi-square test. Results: The synthesized FeSO4 NPs were smaller than 100 nm, and they had more adverse effects on the viability of the HT-29 cells compared to the bulk- FeSO4 at 72 hours. Flow cytometric study showed that the early apoptosis of cells by the bulk form was more than the NPs, but at the low concentration (1.57 mM/mL), the NPs induced more necrosis than the bulk particles (P=0.063). The survival rate of cells facing all concentrations of NPs and bulk- FeSO4 decreased dose dependently (P=0.075). In vivo results revealed that there were no pathological changes in rats’ intestinal tissues. Conclusion: The bulk and NPs of iron have adverse effects on the HT-29 cells, but no histopathological changes were seen on rats’ intestinal cells.

Highlights

Despite the comprehensive application of nanoparticles (NPs) in foods, such as improving flavour and texture, increasing shelf life, enhancing dietary value, the small size of these nanoparticles, i.e. less than 100 nm, and the possibility of their attachment to vital body structures and causing toxic effects, stay a source of concern among scientists, as reflected in the research undertaken [1,2,3,4,5,6,7,8,9,10,11,12]
The particle shape, size and mean diameter of NPs was observed by scanning electron microscopy (SEM; KYKYEM3200, China) and transmission electron microscopy (TEM; Philips CM 10, Eindhoven, The Netherlands)
Cell proliferation and metabolic activity The results showed that HT-29 cells metabolic activity in all the three concentration, except for 6.25 mM/mL of bulk particles, decreased after 72 hours in a dose- and time-dependent manner, so that cell viability at different concentrations of bulk FeSO4 (1.57, 3.125, and 6.25 mM/ mL) was 100%, 96%, and 73.7% at 72 hours, respectively

Summary

Introduction

Despite the comprehensive application of nanoparticles (NPs) in foods, such as improving flavour and texture, increasing shelf life, enhancing dietary value, the small size of these nanoparticles, i.e. less than 100 nm, and the possibility of their attachment to vital body structures and causing toxic effects, stay a source of concern among scientists, as reflected in the research undertaken [1,2,3,4,5,6,7,8,9,10,11,12]. Iron deficiency anemia is a prevalent and Moeini et al widespread nutritional disorder during pregnancy in most countries, including Iran, and a major risk factor for increasing the maternal and neonatal mortality and the prevalence of low birth weight and premature birth (2123) Enriching foods such as bread with iron supplements is an effective and inexpensive strategy for controlling and preventing disorders associated with iron deficiency or iron deficiency compensation; this strategy can cause problems such as changing the taste and color of the food or gastrointestinal tract complications (2426). We made iron (Fe) nanoparticles (NPs) and studied the cytotoxicity of FeSO4 bulk and NPs on HT-29 cells and different doses of these particles on rat intestine.

Methods

Results

Conclusion