Preconditioning of adipose tissue-derived mesenchymal stem cells with deferoxamine increases the production of pro-angiogenic, neuroprotective and anti-inflammatory factors: Potential application in the treatment of diabetic neuropathy

Abstract

Diabetic neuropathy (DN) is one of the most frequent and troublesome complications of diabetes mellitus. Evidence from diabetic animal models and diabetic patients suggests that reduced availability of neuroprotective and pro-angiogenic factors in the nerves in combination with a chronic pro-inflammatory microenvironment and high level of oxidative stress, contribute to the pathogenesis of DN. Mesenchymal stem cells (MSCs) are of great interest as therapeutic agents for regenerative purposes, since they can secrete a broad range of cytoprotective and anti-inflammatory factors. Therefore, the use of the MSC secretome may represent a promising approach for DN treatment. Recent data indicate that the paracrine potential of MSCs could be boosted by preconditioning these cells with an environmental or pharmacological stimulus, enhancing their therapeutic efficacy. In the present study, we observed that the preconditioning of human adipose tissue-derived MSCs (AD-MSCs) with 150μM or 400μM of the iron chelator deferoxamine (DFX) for 48 hours, increased the abundance of the hypoxia inducible factor 1 alpha (HIF-1α) in a concentration dependent manner, without affecting MSC morphology and survival. Activation of HIF-1α led to the up-regulation of the mRNA levels of pro-angiogenic factors like vascular endothelial growth factor alpha and angiopoietin 1. Furthermore this preconditioning increased the expression of potent neuroprotective factors, including nerve growth factor, glial cell-derived neurotrophic factor and neurotrophin-3, and cytokines with anti-inflammatory activity like IL4 and IL5. Additionally, we observed that these molecules, which could also be used as therapeutics, were also increased in the secretome of MSCs preconditioned with DFX compared to the secretome obtained from non-preconditioned cells. Moreover, DFX preconditioning significantly increased the total antioxidant capacity of the MSC secretome and they showed neuroprotective effects when evaluated in an in vitro model of DN. Altogether, our findings suggest that DFX preconditioning of AD-MSCs improves their therapeutic potential and should be considered as a potential strategy for the generation of new alternatives for DN treatment.

Activated mesenchymal stem cell administration inhibits chronic alcohol drinking and suppresses relapse‐like drinking in high‐alcohol drinker rats

“…Human adipose tissue‐derived MSCs (hMSCs) were isolated from fresh subcutaneous adipose tissue samples (abdominal region) obtained from liposuction aspirates of patients undergoing cosmetic liposuction at Clínica Alemana, Santiago, Chile, as previously described by Oses et al . (). A written informed consent was obtained.…”

The Pericytic Phenotype of Adipose Tissue-Derived Stromal Cells Is Promoted by NOTCH2

“…Among these therapeutically relevant cells mesenchymal stem cells (MSC), such as adipose tissue‐derived stromal/stem cells (ASC) , have a strong potency to differentiate into smooth muscle cells and pericytes . Besides this constructive role, ASC are instructive in regenerative processes and secrete a host of mediators that favor the formation and maintenance of blood vessels, that is, vascular endothelial growth factor (VEGF), platelet‐derived growth factor (PDGF), angiopoetins‐1 and 2 (Ang‐1, Ang‐2) . Moreover, ASC promote remodeling of the extracellular matrix , which is essential during vasculogenesis.…”

Cryopreserved H2 O2 -preconditioned human adipose-derived stem cells exhibit fast post-thaw recovery and enhanced bioactivity against oxidative stress

“…Finally, for osteogenic differentiation, cells were seeded at 1,900 cells/cm 2 , and extracellular calcium deposits were qualitatively assessed using Alizarin red S (Sigma, St. Louis, Missouri; Oses et al, ). Samples were fixed in 10% formaldehyde for 10 min at RT, washed with PBS and distilled water, and incubated in 2% Alizarin Red S dye (pH 4.1–4.3) diluted in water for 15 min in the dark with gentle shaking.…”

“…For these reasons, any strategy that improves cell functionality immediately post‐thaw, without the need for an in vitro recovery phase, could be crucial for the success of an off‐the‐shelf cell therapy. We consider that the best strategy is cell in vitro preconditioning before cryopreservation to increase the intrinsic biological activity of ASCs (Baldari et al, ; Boregowda & Phinney, ; Guess et al, ; Oses et al, ), increase stem cell potency, and enhance the regenerative effects of ASC‐based therapies (Amiri, Jahanian‐Najafabadi, & Roudkenar, ). The goal of preconditioning is to induce a beneficial adaptive response in cells, during the stage of in vitro expansion, by exposing them to low levels of a specific stimulus: cytokines, drugs, or various types of stress such as hypoxia, heat shock, nutrient deprivation, acidosis, and/or oxidation (Baldari et al, ; Ezquer, Ezquer, Vicencio, & Calligaris, ; Oses et al, ; Park, Ahn, Sung, Ahn, & Chang, ).…”