Oxidative Stress Response in Adipose-Derived Mesenchymal Stem/Stromal Cells

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20m
ICE Krakow

ICE Krakow

ul. Marii Konopnickiej 17 30-302 Kraków

Speaker

Waheed, Tawakalitu Okikiola (University medical center, Rostock )

Description

OXIDATIVE STRESS RESPONSE IN ADIPOSE-DERIVED MESENCHYMAL STEM/STROMAL CELLS

Tawakalitu Okikiola Waheed, Olga Hahn, Kaarthik Sridharan, Nina Gehm, Caroline Moerke, Kirsten Peters

Introduction: Adipose tissue-derived mesenchymal stem/stromal cells (adMSC) are shown to support wound healing due to their regenerative and immunomodulatory capacities. However, cell survival in the specific wound microenvironment might be affected by oxidative stress, so their ability to regenerate tissues may be impaired. In this study, we investigated the effects of oxidative stress induced by repeated treatment with hydrogen peroxide (H2O2, only briefly stable in medium) or the enzyme glucose oxidase (GOX, which catalyzes the long-term production of H2O2). Using this cell culture model, the short- and long-term effects of oxidative stress on cell survival, proliferation, migration, and differentiation ability were examined.

Methodology: adMSC were isolated from adipose tissue by a CD34-specific isolation protocol. adMSC in passage 4 were exposed to different concentrations of H2O2 or GOX and the viability, proliferation, migration (scratch assay), adipogenic, and osteogenic differentiation capacity of adMSC were examined. At the same time, the extent of oxidative stress was quantified by Amplex Red reagent (detects H2O2) and DCFDA (detects reactive oxygen species/ROS). With this approach, we aimed to relate the induced oxidative stress to the cellular response.

Results: First, we detected the toxicity threshold of the two different oxidative stressors. Only non-toxic concentrations were used for all further investigations. In addition, no significant influence on the metabolic activity of the adMSC could be shown in the amounts of non-toxic H2O2 and GOX used. The adipogenic differentiation capacity (examined over a period of 14 days) was not changed by the non-toxic concentrations of H2O2 or GOX. The degree of osteogenic differentiation (examined over a period of 35 days) was under the influence of higher oxidative stress (0.5 mM and 1 mM H2O2, respectively 5 mU/ml and 10 mU/ml GOX) due to an increase in extracellular calcium deposition. Initial investigations show that the migration activity (detected using the scratch assay) is significantly increased by the addition of non-toxic amounts of H2O2 and GOX.

Conclusion: These results indicate a certain resilience of adMSC to oxidative stress. We could show that osteogenic differentiation and migration ability are increased by non-toxic concentrations of H2O2 and GOX. We are currently investigating how oxidative stress affects the inflammatory response of cells. For this purpose, cytokines (e.g. IL-6) and growth factors (e.g. TGF-β, bFGF) important for wound healing are examined. These fundamental investigations can help to describe the therapeutic potential of adMSC in the wound healing process more precisely and, if necessary, to influence them.

Keywords: Mesenchymal stem/stromal cells, hydrogen peroxide, glucose oxidase, oxidative stress, wound healing.

References:
1. Atashi, Fatemeh. et al., Stem cells and development. 24(10), 1150-63 (2015).
2. Rochette, L. et al., International Journal of Molecular Sciences. 21(23), 9262 (2020).
3. Guo, L. et al., Stem Cell Research and Therapy. 11(1), 434 (2020).
4. Dikalov, S.I. and Harrison D.G. Antioxidants and Redox Signaling. 20(2), 372-82 (2014).
5. Milton, R.D. et al., Physical Chemistry Chemical Physics. 15, 19371-19379 (2013)

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