Human adipose-derived stem cells obtained from lipoaspirates are highly susceptible to hydrogen peroxide mediated cytogenotoxicity

Alencar Kolinski Machado; Sabrina Guastavino Homrich; Cíntia Corte Real Rodrigues; Verônica Farina Azzolin; Marta Maria Medeiros Frescura Duarte; Danieli Monteiro Pillar; Luana Suéling Lenz; Tais Cristina Unfer; Diulie Valente de Souza; Ivana Beatrice Mânica da Cruz

doi:10.18593/abh.17060

Authors

Alencar Kolinski Machado Franciscan University https://orcid.org/0000-0003-2003-8420
Sabrina Guastavino Homrich Federal University of Santa Maria https://orcid.org/0000-0002-8202-3382
Cíntia Corte Real Rodrigues Federal University of Rio Grande do Sul https://orcid.org/0000-0002-7889-5911
Verônica Farina Azzolin Federal University of Santa Maria https://orcid.org/0000-0002-8191-5450
Marta Maria Medeiros Frescura Duarte Federal University of Santa Maria https://orcid.org/0000-0003-4371-4989
Danieli Monteiro Pillar Federal University of Santa Maria https://orcid.org/0000-0001-9760-7586
Luana Suéling Lenz Federal University of Rio Grande do Sul https://orcid.org/0000-0001-6346-4265
Tais Cristina Unfer Federal University of Sergipe https://orcid.org/0000-0001-6725-4003
Diulie Valente de Souza https://orcid.org/0000-0002-6126-2501
Ivana Beatrice Mânica da Cruz Federal University of Santa Maria https://orcid.org/0000-0002-0310-3190

DOI:

https://doi.org/10.18593/abh.17060

Keywords:

Adult stem cells genotoxicity, Oxidative stress, Lipoaspirates

Abstract

There is evidence that H₂O₂ can induce the proliferation, migration, and regeneration of stem cells, as well as that of adipose-derived stem cells (ASCs). This could be useful to expand the possible uses of ASCs in therapeutic applications. However, the safety profile of H₂O₂use in stem cells is not clear yet. Therefore, the present study evaluated the acute cytotoxic, oxidative and genotoxic effects of different concentrations of H₂O₂ on ASCs obtained from human lipoaspirates. The ASCs were treated with 1–1000 μM H₂O₂for two hours. Cell viability was evaluated by double-strand DNA determination. Apoptosis induction was analyzed measuring active levels of caspases 1, 3 and 8. Biochemical oxidative stress markers were analyzed and genotoxic effects were assessed by DNA comet assay. All H₂O₂concentrations increased ASC mortality rates with approximately 100% mortality achieved at ≥ 200 μM. Active caspases 1, 3 and 8, oxidative stress, as well as oxidative damage as assessed by lipid peroxidation increased dose‐dependently. There was also an approximate 50% increase in catalase levels in cells exposed to all H₂O₂tested concentrations. H₂O₂ concentrations of ≥ 10 μM were genotoxic. These results suggest that ASCs are highly sensitive to H₂O₂ exposition. In addition, DNA damage in the surviving cells may affect their proliferative and differentiation capacity, as well as their safety profile for therapeutic use.

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References

Fernandéz Vallone VB, Romaniuk MA, Choi H, Labovsky V, Otaegui J, Chasseing NA. Mesenchymal stem cells and their use in therapy: what has been achieved? Differentiation 2013; 85:1-2. doi: 10.1016/j.diff.2012.08.004

Ong WK, Sugii S. Adipose-derived stem cells: Fatty potentials for therapy. Int J Biochem Cell Biol. 2013; 45(6):1083-1086. doi: 10.1016/j.biocel.2013.02.013

Lindroos B, Suuronen R, Miettinen S. The potential of adipose stem cells in regenerative medicine. Stem Cell Rev. 2011; 7(2):269-91. doi: 10.1007/s12015-010-9193-7

Park SG, Kim J, Xia Y, Sung J. Generation of reactive oxygen species in adipose-derived stem cells: Friend or Foe? Expert Opin Ther Targets. 2011; 15(11):1297-306. doi: 10.1517/14728222.2011.628315

Urao N, Ushio-Fukai M. Redox regulation of stem/progenitor cells and boné marrow niche. Free Radic Biol Med. 2013; 54:26-39. doi: 10.1016/j.freeradbiomed.2012.10.532

Zhang J, Chen G, Wang Y, Zhao J, Duan H, Liao LM, et al. Hydrogen peroxide preconditioning enhances the therapeutic efficacy of Wharton’s Jelly mesenchymal stem cells after myocardial infarction. Chin. Med. J. 2012; 125(19):3472-8. PMID: 23044308.

Robaszkiewicz A, Erdélyi K, Kovács K, Kovács I, Bai P, Rajnavölgyi E, et al. Hydrogen peroxide-induced poly(ADP-ribosyl)ation regulates osteogenic differentiation-associated cell death. Free Radic Biol Med. 2012; 53(8):1552-64. doi: 10.1016/j.freeradbiomed.2012.08.567

Burova E, Borodkina A, Shatrova A, Nikolsky N. Sublethal oxidative stress induces the premature senescence of human mesenchymal stem cells derived from endometrium. Oxid Med Cell Longev. 2013: 1-12. doi: 10.1155/2013/474931

Buehrer BM, Cheatham B. Isolation and characterization of human adipose-derived stem cells for use in tissue engineering. Methods Mol Biol. 2013; 1001:1-11. doi: 10.1007/978-1-62703-363-3_1

Costa F, Dornelles E, Manica-Cattani MF, Algarve T, Souza-Filho O, Sagrillo M, et al. Influence of Val16Ala SOD2 polymorphism on the in-vitro effect of clomiphene citrate in oxidative metabolism. Reprod biomed online 2012; 24:474-81. doi: 10.1016/j.rbmo.2012.01.009

Ohkawa H, Ohishi H, Yagi K. Assay for lipid peroxide in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 1979; 95:351-8. doi: 10.1016/0003-2697(79)90738-3

Boveris A, Cadenas E. Cellular source and steady-state levels of reactive oxygen species. In: Mazzaro D, Clerch L, editors. Oxygen, Gene Expression and Cellular Function. New York: Marcel Dekker Inc; 1997.

Aebi H. Catalase in vitro. Methods Enzymol 1984; 105:121-7. doi:10.1016/S0076-6879(84)05016-3

Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988; 175(1):184-91. PMID: 3345800

Nadin SB, Vargas-Roig LM, Ciocca DRA. A silver staining method for single-cell gel assay. J Histochem Cytochem 2001; 49(9):1183-6. doi: 10.1177/002215540104900912

Sherman MH, Bassing CH, Teitell MA. (2011) Regulation of cell differentiation by the DNA damage response. Trends Cell Biol 2011; 21(5):312-9. doi: 10.1016/j.tcb.2011.01.004

Weiss CN, Ito K. DNA damage response, redox status and hematopoiesis. Blood Cells Mol Dis 2014; 52(1):12-8. doi: 10.1016/j.bcmd.2013.08.002

Naka K, Muraguchi T, Hoshii T, Hirao A. (2008) Regulation of reactive oxygen species and genomic stability in hematopoietic stem cells. Antioxid Redox Signal 2008; 10(11):1883-94. doi: 10.1089/ars.2008.2114

Sardina JL, Lopez-Ruano G, Sanchez-Sanchez B, Llanillo M, Hernández-Hernández A. Reactive oxygen species: are they important for haematopoiesis? Crit Rev Oncol Hematol. 2012; 81(3):257-74. doi: 10.1016/j.critrevonc.2011.03.005

Jang YY, Sharkis SJ. A low level of reactive oxygen species selects for primitive hematopoietic stem cells that may reside in the low-oxygenic niche. Blood 2007; 110(8):3056-63. doi: 10.1182/blood-2007-05-087759

Groeger G, Quiney C, Cotter TG. Hydrogen peroxide as a cell-survival signaling molecule. Antioxid Redox Signal 2009; 11(11):2655-71. doi: 10.1089/ARS.2009.2728

Chaudhari P, Ye Z, Jang YY. Roles of reactive oxygen species in the fate of stem cells. Antioxid Redox Signal 2014; 20(12):1881-90. doi: 10.1089/ars.2012.4963

Mooney BM, Raof NA, Li Y, Xie Y. Convergent mechanisms in pluripotent stem cells and cancer: implications for stem cell engineering. Biotechnol J. 2013; 8(4):408-19. doi: 10.1002/biot.201200202

Sosa V, Moliné T, Somoza R, Paciucci R, Kondoh H, LLeonart ME. Oxidative stress and cancer: an overview. Ageing Res Rev. 2012; 12(1):376-90. doi: 10.1016/j.arr.2012.10.004

Human adipose-derived stem cells obtained from lipoaspirates are highly susceptible to hydrogen peroxide mediated cytogenotoxicity

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