Up-Regulation of Superoxide Dismutase 2 in 3D Spheroid Formation Promotes Therapeutic Potency of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells

Umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) are accessible, available in abundance, and have been shown to be a promising source that can regenerate cartilage in patients with osteoarthritis or other orthopedic diseases. Recently, a three-dimensional (3D) cell culture system was developed to mimic the naive tissue microenvironment. However, the efficacy of cells generated from the 3D spheroid culture system has not yet been elucidated. In the present study, we demonstrate the changes in superoxide dismutase 2 (SOD2) gene expression, an indicator of oxidative stress, on 3D spheroid MSCs. Moreover, siRNA transfection and neutralizing antibody investigations were performed to confirm the function of SOD2 and E-cadherin. Overall, we found that SOD2 siRNA transfection in the spheroid form of MSCs increases the expression of apoptotic genes and decreases the clearance of mitochondrial reactive oxygen species (ROS). As a result, we confirm that 3D spheroid formation increases E-cadherin and SOD2 expression, ultimately regulating the phosphoinositide 3-kinase (PI3K/pAkt/pNrf2 and pERK/pNrf2 signaling pathway. Additionally, we show that SOD2 expression on 3D spheroid MSCs affects the regeneration rates of destructive cartilage in an osteoarthritic model. We postulate that the impact of SOD2 expression on 3D spheroid MSCs reduces oxidative stress and apoptosis, and also promotes cartilage regeneration.

[1]  Ya-Wen Chen,et al.  Arsenic induces human chondrocyte senescence and accelerates rat articular cartilage aging , 2019, Archives of Toxicology.

[2]  Yehong Yang,et al.  Protective Effects Of Astragalus Polysaccharides On Oxidative Stress In High Glucose-Induced Or SOD2-Silenced H9C2 Cells Based On PCR Array Analysis , 2019, Diabetes, metabolic syndrome and obesity : targets and therapy.

[3]  W. Oh,et al.  Decorin Secreted by Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells Induces Macrophage Polarization via CD44 to Repair Hyperoxic Lung Injury , 2019, International journal of molecular sciences.

[4]  Eun-Ji Choi,et al.  Intra-articular delivery of synovium-resident mesenchymal stem cells via BMP-7-loaded fibrous PLGA scaffolds for cartilage repair. , 2019, Journal of controlled release : official journal of the Controlled Release Society.

[5]  E. Stelzer,et al.  E-cadherin, actin, microtubules and FAK dominate different spheroid formation phases and important elements of tissue integrity , 2018, Biology Open.

[6]  Min Zhao,et al.  Protective effects of SOD2 overexpression in human umbilical cord mesenchymal stem cells on lung injury induced by acute paraquat poisoning in rats , 2018, Life sciences.

[7]  Haniye Sadat Sajadi,et al.  Global, regional, and national incidence, prevalence, and years lived with disability for 354 diseases and injuries for 195 countries and territories, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017 , 2018, The Lancet.

[8]  S. Nagata.  Apoptosis and Clearance of Apoptotic Cells. , 2018, Annual review of immunology.

[9]  L. Baptista,et al.  Adult Stem Cells Spheroids to Optimize Cell Colonization in Scaffolds for Cartilage and Bone Tissue Engineering , 2018, International journal of molecular sciences.

[10]  X. Lei,et al.  Spheroid-cultured human umbilical cord-derived mesenchymal stem cells attenuate hepatic ischemia-reperfusion injury in rats , 2018, Scientific Reports.

[11]  R. Oberley-Deegan,et al.  Differential Effects of Superoxide Dismutase Mimetics after Mechanical Overload of Articular Cartilage , 2017, Antioxidants.

[12]  S. Eskelinen,et al.  Correlation between E-cadherin interactions, survivin expression, and apoptosis in MDCK and ts-Src MDCK cell culture models. , 2017, Laboratory investigation; a journal of technical methods and pathology.

[13]  Mei Zhao,et al.  Inflammation-mediated SOD-2 upregulation contributes to epithelial-mesenchymal transition and migration of tumor cells in aflatoxin G1-induced lung adenocarcinoma , 2017, Scientific Reports.

[14]  LiuLi-min,et al.  In Situ Release of VEGF Enhances Osteogenesis in 3D Porous Scaffolds Engineered with Osterix-Modified Adipose-Derived Stem Cells. , 2017, Tissue engineering. Part A.

[15]  Eva Syková,et al.  The therapeutic potential of three-dimensional multipotent mesenchymal stromal cell spheroids , 2017, Stem Cell Research & Therapy.

[16]  C. V. van Blitterswijk,et al.  Cell aggregation enhances bone formation by human mesenchymal stromal cells. , 2017, European cells & materials.

[17]  D. McDermott,et al.  Mechanisms of Sustained Neutrophilia in Patient WHIM-09, Cured of WHIM Syndrome by Chromothripsis , 2017, Journal of Clinical Immunology.

[18]  L. Poulain,et al.  Chondrogenic commitment of human umbilical cord blood-derived mesenchymal stem cells in collagen matrices for cartilage engineering , 2016, Scientific Reports.

[19]  R. Mahla.  Stem Cells Applications in Regenerative Medicine and Disease Therapeutics , 2016, International journal of cell biology.

[20]  B. Jiang,et al.  Spheroid Mesenchymal Stem Cells and Mesenchymal Stem Cell-Derived Microvesicles: Two Potential Therapeutic Strategies. , 2016, Stem cells and development.

[21]  Zoe Cesarz,et al.  Spheroid Culture of Mesenchymal Stem Cells , 2015, Stem cells international.

[22]  M. Detamore,et al.  Stem Cells in Aggregate Form to Enhance Chondrogenesis in Hydrogels , 2015, PloS one.

[23]  Michelle R. Campbell,et al.  Beyond antioxidant genes in the ancient Nrf2 regulatory network. , 2015, Free radical biology & medicine.

[24]  S. Holley,et al.  Integration of cell-cell and cell-ECM adhesion in vertebrate morphogenesis. , 2015, Current opinion in cell biology.

[25]  T. Talaei-Khozani,et al.  An in vitro model for hepatocyte-like cell differentiation from Wharton’s jelly derived-mesenchymal stem cells by cell-base aggregates , 2015, Gastroenterology and hepatology from bed to bench.

[26]  T. Sasho,et al.  Mechanical overloading causes mitochondrial superoxide and SOD2 imbalance in chondrocytes resulting in cartilage degeneration , 2015, Scientific Reports.

[27]  J. Bao,et al.  Three-dimensional spheroid culture of human umbilical cord mesenchymal stem cells promotes cell yield and stemness maintenance , 2015, Cell and Tissue Research.

[28]  E. Macit,et al.  The relation between oxidative stress, inflammation, and neopterin in the paraquat-induced lung toxicity , 2015, Human & experimental toxicology.

[29]  S. Grandemange,et al.  Manganese superoxide dismutase in breast cancer: from molecular mechanisms of gene regulation to biological and clinical significance. , 2014, Free radical biology & medicine.

[30]  Yaojiong Wu,et al.  Epigenetic changes of mesenchymal stem cells in three-dimensional (3D) spheroids , 2014, Journal of cellular and molecular medicine.

[31]  Rachelle Buchbinder,et al.  Burden of disability due to musculoskeletal (MSK) disorders. , 2014, Best practice & research. Clinical rheumatology.

[32]  D. Candas,et al.  MnSOD in oxidative stress response-potential regulation via mitochondrial protein influx. , 2014, Antioxidants & redox signaling.

[33]  J. Karp,et al.  Mesenchymal stem cells: immune evasive, not immune privileged , 2014, Nature Biotechnology.

[34]  D. Prockop,et al.  Dynamic compaction of human mesenchymal stem/precursor cells into spheres self‐activates caspase‐dependent IL1 signaling to enhance secretion of modulators of inflammation and immunity (PGE2, TSG6, and STC1) , 2013, Stem cells.

[35]  Jerry C. Hu,et al.  Chondrogenically Tuned Expansion Enhances the Cartilaginous Matrix-Forming Capabilities of Primary, Adult, Leporine Chondrocytes , 2013, Cell transplantation.

[36]  Jerry C. Hu,et al.  Unlike Bone, Cartilage Regeneration Remains Elusive , 2012, Science.

[37]  Byung-Soo Kim,et al.  Enhanced cartilage formation via three-dimensional cell engineering of human adipose-derived stem cells. , 2012, Tissue engineering. Part A.

[38]  Byung-Soo Kim,et al.  Transplantation of cord blood mesenchymal stem cells as spheroids enhances vascularization. , 2012, Tissue engineering. Part A.

[39]  E. J. Lee,et al.  Spherical bullet formation via E-cadherin promotes therapeutic potency of mesenchymal stem cells derived from human umbilical cord blood for myocardial infarction. , 2012, Molecular therapy : the journal of the American Society of Gene Therapy.

[40]  Peter G. Schultz,et al.  A Stem Cell–Based Approach to Cartilage Repair , 2012, Science.

[41]  P. Wilder-Smith,et al.  Three-dimensional spheroid culture of human gingiva-derived mesenchymal stem cells enhances mitigation of chemotherapy-induced oral mucositis. , 2012, Stem cells and development.

[42]  Yuan-Yu Hsueh,et al.  Spheroid Formation and Neural Induction in Human Adipose-Derived Stem Cells on a Chitosan-Coated Surface , 2012, Cells Tissues Organs.

[43]  T. Young,et al.  The influence of spheroid formation of human adipose-derived stem cells on chitosan films on stemness and differentiation capabilities. , 2012, Biomaterials.

[44]  Liming Bian,et al.  Enhanced MSC chondrogenesis following delivery of TGF-β3 from alginate microspheres within hyaluronic acid hydrogels in vitro and in vivo. , 2011, Biomaterials.

[45]  H. Thielecke,et al.  A scaffold-free in vitro model for osteogenesis of human mesenchymal stem cells. , 2011, Tissue & cell.

[46]  Nikolay Bazhanov,et al.  Aggregation of human mesenchymal stromal cells (MSCs) into 3D spheroids enhances their antiinflammatory properties , 2010, Proceedings of the National Academy of Sciences.

[47]  T. Kirkwood,et al.  Superoxide dismutase downregulation in osteoarthritis progression and end-stage disease , 2010, Annals of the rheumatic diseases.

[48]  Weihua Yu,et al.  Chondrogenic differentiation of human mesenchymal stem cells: a comparison between micromass and pellet culture systems , 2010, Biotechnology Letters.

[49]  B. Zhu,et al.  Mitochondrial superoxide dismutase SOD2, but not cytosolic SOD1, plays a critical role in protection against glutamate-induced oxidative stress and cell death in HT22 neuronal cells. , 2010, Free radical biology & medicine.

[50]  J. Mateos,et al.  Mitochondrial Dysregulation of Osteoarthritic Human Articular Chondrocytes Analyzed by Proteomics , 2009, Molecular & Cellular Proteomics.

[51]  G. Berx,et al.  The cell-cell adhesion molecule E-cadherin , 2008, Cellular and Molecular Life Sciences.

[52]  E. Lander,et al.  Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. , 2008, Cancer research.

[53]  Michael R Rosen,et al.  Mesenchymal Stem Cells Support Migration, Extracellular Matrix Invasion, Proliferation, and Survival of Endothelial Cells In Vitro , 2007, Stem cells.

[54]  F. Blanco,et al.  Cell death and apoptosis in osteoarthritic cartilage. , 2007, Current drug targets.

[55]  Masayuki Yamamoto,et al.  Nrf2-Keap1 defines a physiologically important stress response mechanism. , 2004, Trends in molecular medicine.

[56]  Joseph A. Buckwalter,et al.  Aging, articular cartilage chondrocyte senescence and osteoarthritis , 2004, Biogerontology.

[57]  T. Aw,et al.  Differential Effects of Superoxide Dismutase Isoform Expression on Hydroperoxide-induced Apoptosis in PC-12 Cells* , 2003, The Journal of Biological Chemistry.

[58]  Hans Hauner,et al.  Cartilage-like gene expression in differentiated human stem cell spheroids: a comparison of bone marrow-derived and adipose tissue-derived stromal cells. , 2003, Arthritis and rheumatism.

[59]  Yong-jie Wu,et al.  Anti-inflammatory effect of recombinant human superoxide dismutase in rats and mice and its mechanism. , 2002, Acta pharmacologica Sinica.

[60]  T. Aigner,et al.  Apoptotic cell death is not a widespread phenomenon in normal aging and osteoarthritis human articular knee cartilage: a study of proliferation, programmed cell death (apoptosis), and viability of chondrocytes in normal and osteoarthritic human knee cartilage. , 2001, Arthritis and rheumatism.