Elsevier

Cytotherapy

Volume 16, Issue 6, June 2014, Pages 764-775
Cytotherapy

Original paper
Mesenchymal stromal cells
Mesenchymal stromal cell injection protects against oxidative stress in Escherichia coli–induced acute lung injury in mice

https://doi.org/10.1016/j.jcyt.2013.12.006Get rights and content

Abstract

Background aims

Stem cells may be a promising therapy for acute respiratory distress syndrome. Recent in vivo and in vitro studies suggested that the mesenchymal stromal cells (MSCs) have anti-oxidative stress properties. We hypothesized that intravenous injection of bone marrow–derived mesenchymal stem cells (MSCs) could attenuate Escherichia coli–induced acute lung injury (ALI) in mice by controlling the oxidative stress status.

Methods

Eighty mice were randomly divided into four groups: group 1 (control group) received 25 μL of saline as a vehicle; group 2 contained E coli–induced ALI mice; group 3 included mice that received MSCs before induction of ALI; group 4 included mice that received MSCs after induction of ALI. Lung samples were isolated and assayed for oxidative stress variables and histopathologic analysis. Total anti-oxidant capacity was measured in broncho-alveolar lavage.

Results

Pre- and post-injury MSC injection increased survival, reduced pulmonary edema and attenuated lung injuries in ALI mice. Histologically, MSCs exhibited a considerable degree of preservation of the pulmonary alveolar architecture. An increase of anti-oxidant enzyme activities and a decrease of myeloperoxidase activity and malondialdehyde levels in the MSC recipient groups versus the ALI group were found. Furthermore, the total anti-oxidant capacity and reduced glutathione levels were significantly increased in MSCs recipient groups versus the ALI group. Weak +ve inducible nitric oxide synthase immuno-expression in groups that received MSCs was detected. Pre-injury MSC injection showed better effects than did post-injury MSC injection.

Conclusions

Systemic bone marrow–derived MSC injection was effective in modulating the oxidative stress status in E coli–induced acute lung injury in mice.

Introduction

Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS), are lung diseases characterized by a severe inflammatory process causing alveolar damage and resulting in a variable degree of ventilation perfusion mismatch, severe hypoxemia, poor lung compliance and noncardiogenic pulmonary edema (1). Patients with ALI have a ratio of arterial oxygen tension to fraction of inspired oxygen (PaO2/FiO2) of 201 to 300 mm Hg, whereas patients with ARDS have worse hypoxemia, with a PaO2/FiO2 of ≤200 mm Hg (2). Acute respiratory distress syndrome is a major cause of acute respiratory failure in critically ill patients. Several etiological factors associated with the development of ALI as sepsis, pneumonia and trauma with multiple transfusions accounting for most cases. A major cause for the development of ALI is sepsis, wherein Gram-negative bacteria are a prominent cause (3). There are no therapies for ARDS until now, yet management remains supportive (2). Improvements in these measures, such as protective mechanical ventilation strategies (4), restrictive intravenous fluid management (5) and prone positioning of severely hypoxemic patients (6) have decreased mortality rates from ARDS. Despite all innovations in intensive care medicine, the mortality of acute respiratory failure from ARDS remains up to 40% (1). More complex strategies, aimed to reduce early injury and/or enhance repair after injury, are needed.

These insights have led to a renewed interest in the therapeutic potential of mesenchymal stromal cells (MSCs). MSCs are multipotent cells derived from adult tissues that are capable of self-renewal and differentiation into chondrocytes, osteocytes and adipocytes. The derivation of MSCs from adult tissues, their relative ease of isolation and enormous expansion potential in culture make them attractive therapeutic candidates (7). They are immunologically well tolerated (8) and therefore can be transplanted from an individual to another, an important advantage for acute illnesses such as ARDS. Recent studies have demonstrated that bone marrow (BM)-derived MSCs reduce lung injury in experimental models of lipopolysaccharide (LPS)-induced ALI in mouse 9, 10, 11, even though the mechanisms underlying the therapeutic effect of MSCs on ALI has yet to be elucidated. One of the main mechanisms involved in the pathogenesis of ALI is through an increase in oxidative stress, resulting from an imbalance between the production of reactive oxygen species (ROS) and their elimination (12). Intracellular ROS accumulation can result in nitration and/or oxidation of cellular biomolecules including proteins, lipids and DNA (12). Therefore, the anti-oxidative enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) can prevent lung injury induced by LPS from ROS exposure (13).

We hypothesized that intravenous injection of BM-MSCs could attenuate Escherichia coli–induced ALI in mice, and, if so, the protective mechanism may be mediated by controlling the oxidative stress status. We examined the effects of MSCs in two groups, one with the injection of MSCs before injury and the other after ALI injury, to evaluate whether MSCs had a protective rather than a therapeutic effect. The anti-oxidant markers as SOD, CAT, GPx and glutathione reductase (GR) activities, reduced glutathione (GSH) levels in lung homogenates and total anti-oxidant capacity (TAC) in broncho-alveolar lavage (BAL) were estimated; malondialdehyde (MDA) level as an indicator of lipid peroxidation, myeloperoxidase (MPO) activity as an indicator of neutrophils recruitment and inducible nitric oxide synthase (iNOS) expression were evaluated to elucidate the potential anti-oxidative responses of the MSCs.

Section snippets

Methods

This study was performed in a stem cell research laboratory in the Medical Biochemistry Department in collaboration with Clinical Pharmacology, Histology and Cell Biology, Microbiology and Immunology, and Anesthesia and Intensive Care Departments, Faculty of Medicine, Zagazig University, Egypt.

MSC characterization

Mouse MSCs were obtained from bone marrow of Balb c mice by adherence to plastic culture flasks. Morphologically, these cells had a spindled, fibroblast appearance after expansion. Cells that had been passaged four times were used in all experiments. Flow cytometric analysis demonstrated that MSCs did not express CD34 (0.2%) but expressed CD105, CD73 (99.5% and 96.4%, respectively).

Survival rate

E coli–induced ALI significantly reduced the survival rate (70%, 14/20 mice surviving) at post-injury day 2

Discussion

ALI/ARDS account for 10–15% intensive care unit admissions; even with the current advances in lung-protective ventilation and fluid management, there is still a high mortality rate (29). They are followed by poor survival and diminished quality of lung function. In addition, survivors often have long-term neuromuscular, cognitive and psychological symptoms (2). Probable reasons for the failure to find a successful therapy include deficits in our understanding of the disease, coupled with a

Conclusions

Our study suggests that BM-MSC injection in mice with E coli–induced ALI improves survival and attenuates ALI through anti-oxidative mechanisms. To date, although few mesenchymal stem cells clinical trials in critically ill patients have not reported adverse immune side effects; the reported data were very limited. Therefore, Mesenchymal stem cells may be a promising therapy for patients suffering from ARDS but the gaps in our knowledge regarding the optimal mesenchymal stem cell

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