Abstract

This in vitro study aimed to find the best method of granulocyte isolation for subsequent labeling with multimodal nanoparticles (magnetic and fluorescent properties) to enable detection by optical and magnetic resonance imaging (MRI) techniques. The granulocytes were obtained from venous blood samples from 12 healthy volunteers. To achieve high purity and yield, four different methods of granulocyte isolation were evaluated. The isolated granulocytes were labeled with multimodal superparamagnetic iron oxide nanoparticles (M-SPIONs) coated with dextran, and the iron load was evaluated qualitatively and quantitatively by MRI, near-infrared fluorescence (NIRF) and inductively coupled plasma mass spectrometry (ICP-MS). The best method of granulocyte isolation was Percoll with Ficoll, which showed 95.92% purity and 94% viability. After labeling with M-SPIONs, the granulocytes showed 98.0% purity with a yield of 3.5 × 106 cells/mL and more than 98.6% viability. The iron-loading value in the labeled granulocytes, as obtained by MRI, was 6.40 ± 0.18 pg/cell. Similar values were found with the ICP-MS and NIRF imaging techniques. Therefore, our study shows that it is possible to isolate granulocytes with high purity and yield and labeling with M-SPIONs provides a high internalized iron load and low toxicity to cells. Therefore, these M-SPION-labeled granulocytes could be a promising candidate for future use in inflammation/infection detection by optical and MRI techniques.

Highlights

  • Inflammation is the immune system0 s response to pathological agents, damaged cells and toxins, and it aims to maintain integrity, defense against microorganisms and cellular and tissue repair [1,2,3]

  • Granulocytes were isolated by four different methods: method I—Percoll gradients, method

  • Granulocytes are known for their high potential and efficiency in detecting and eradicating microbial infections through an immune response against invading pathogens [66]

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Summary

Introduction

Inflammation is the immune system0 s response to pathological agents (infection), damaged cells and toxins, and it aims to maintain integrity, defense against microorganisms and cellular and tissue repair [1,2,3]. Acute inflammation of the lower respiratory tract is a leading cause of mortality and morbidity worldwide, with the greatest prevalence in children younger than five years [4,5,6]. Sepsis is another important public health problem; it occurs in 30% to 50% of hospitalizations that culminate in death [7,8] due to exacerbated and inappropriate inflammation, and it can cause septic shock and organ failure [9]. Hard-to-reach or occult inflammation and infection processes are clinically challenging. Accurate and early diagnosis of inflammation and infection helps reduce mortality and morbidity and increase drug treatment success [19,20,21]

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