Ouabain induces in vitro differentiation of human bone marrow mesenchymal stem cells into osteoblasts

doi:10.12307/2022.556

Abstract

Abstract:

BACKGROUND: As a ligand of Na+/K+-ATPase, ouabain can be combined with it to regulate cell proliferation and metabolism. Studying the effect of ouabain on the osteogenic differentiation of bone marrow mesenchymal stem cells can provide ideas for the treatment of bone defects and bone diseases.
OBJECTIVE: To explore the effects of different concentrations of Na+/K+-ATPase ligand ouabain on the osteogenic differentiation of human bone marrow mesenchymal stem cells.
METHODS: Human bone marrow mesenchymal stem cells were isolated from fresh bone marrow samples from patients that experienced hip replacement, using the whole bone marrow direct adherence method, as experimental cells for inducing osteogenic differentiation. Before osteoinduction, human bone marrow mesenchymal stem cells were treated in different concentrations of ouabain (10, 100, and 1 000 nmol/L) for 24 hours. The untreated human bone marrow mesenchymal stem cells were used as the control group to induce directly. At 21 days after osteoinduction, alizarin red staining was performed. The proportion area was measured using Image Pro Plus software to compare the differences in the amount of calcified nodules produced in each group.
RESULTS AND CONCLUSION: Compared with the control group, as human bone marrow mesenchymal stem cells in vitro treated with ouabain in low concentration (10 nmol/L), the number of calcified nodules increased after alizarin red staining and the staining areas were higher than that in the control group (P < 0.05). The number of calcified nodules observed after treatment in the high concentration group (100, 1 000 nmol/L) was reduced, and the measured stained area was smaller than that in the control group (P < 0.05). The higher the concentration, the fewer the calcifiednodules were. It is concluded that the fresh bone marrow form hip arthroplasty can be used as a test material for isolating human bone marrow mesenchymal stem cells. Low concentrations of ouabain promote differentiation of human bone marrow mesenchymal stem cells into osteoblasts, while a high concentration of ouabain showed an inhibitory effect.

Key words: stem cells, bone marrow, human bone marrow mesenchymal stem cells, ouabain, Na+/K+-ATPase, osteogenic differentiation, artificial hip replacement

CLC Number:

Cui Yinpeng, Guo Ai, Ma Lifeng, Liu Zhenjiang. Ouabain induces in vitro differentiation of human bone marrow mesenchymal stem cells into osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(24): 3796-3801.

Figures/Tables 8

References

[1] MEPPELINK AM, WANG XH, BRADICA G, et al. Rapid isolation of bone marrow mesenchymal stromal cells using integrated centrifuge-based technology. Cytotherapy. 2016;18(6):729-739.
[2] JOHNSTONE BH, MILLER HM, BECK MR, et al. Identification and characterization of a large source of primary mesenchymal stem cells tightly adhered to bone surfaces of human vertebral body marrow cavities. Cytotherapy. 2020;22(11):617-628.
[3] MAHBOUDI H, KAZEMI B, SOLEIMANI M, et al. Enhanced chondrogenesis of human bone marrow mesenchymal Stem Cell (BMSC) on nanofiber-based polyethersulfone (PES) scaffold. Gene. 2018;643:98-106.
[4] RATH SN, NOOEAID P, ARKUDAS A, et al. Adipose- and bone marrow-derived mesenchymal stem cells display different osteogenic differentiation patterns in 3D bioactive glass-based scaffolds. J Tissue Eng Regen Med. 2016;10(10):E497-E509.
[5] HUANG GT, GRONTHOS S, SHI S. Mesenchymal stem cells derived from dental tissues vs. those from other sources: their biology and role in regenerative medicine. J Dent Res. 2009;88(9):792-806.
[6] CENTENO CJ, BUSSE D, KISIDAY J, et al. Increased knee cartilage volume in degenerative joint disease using percutaneously implanted, autologous mesenchymal stem cells. Pain Physician. 2008;11(3):343-353.
[7] SHAO X, GOH JC, HUTMACHER DW, et al. Repair of large articular osteochondral defects using hybrid scaffolds and bone marrow-derived mesenchymal stem cells in a rabbit model. Tissue Eng. 2006;12(6):1539-1551.
[8] MORENO C, YANO S, BEZANILLA F, et al. Transient Electrical Currents Mediated by the Na+/K+-ATPase: A Tour from Basic Biophysics to Human Diseases. Biophys J. 2020;119(2):236-242.
[9] LI Z, CAI T, TIAN J, et al. NaKtide, a Na/K-ATPase-derived peptide Src inhibitor, antagonizes ouabain-activated signal transduction in cultured cells. J Biol Chem. 2009;284(31):21066-21076.
[10] GABLE ME, ABDALLAH SL, NAJJAR SM, et al. Digitalis-induced cell signaling by the sodium pump: on the relation of Src to Na(+)/K(+)-ATPase. Biochem Biophys Res Commun. 2014;446(4):1151-1154.
[11] HAMLYN JM, BLAUSTEIN MP, BOVA S, et al. Identification and characterization of a ouabain-like compound from human plasma. Proc Natl Acad Sci U S A. 1991;88(14):6259-6263.
[12] OTSU K, KURUMA A, YANAGIDA E, et al. Na+/K+ ATPase and its functional coupling with Na+/Ca2+ exchanger in mouse embryonic stem cells during differentiation into cardiomyocytes. Cell Calcium. 2005;37(2):137-151.
[13] LEE YK, NG KM, LAI WH, et al. Ouabain facilitates cardiac differentiation of mouse embryonic stem cells through ERK1/2 pathway. Acta Pharmacol Sin. 2011;32(1):52-61.
[14] SAYED M, DRUMMOND CA, EVANS KL, et al. Effects of Na/K-ATPase and its ligands on bone marrow stromal cell differentiation. Stem Cell Res. 2014;13(1):12-23.
[15] TIAN J, LI X, LIANG M, et al. Changes in sodium pump expression dictate the effects of ouabain on cell growth. J Biol Chem. 2009;284(22): 14921-14929.
[16] MAKIHIRA S, NIKAWA H, KAJIYA M, et al. Blocking of sodium and potassium ion-dependent adenosine triphosphatase-α1 with ouabain and vanadate suppresses cell-cell fusion during RANKL-mediated osteoclastogenesis. Eur J Pharmacol. 2011;670(2-3):409-418.
[17] 潘欣宇,崔颖,马林祥,等.骨髓间充质干细胞的研究进展及临床应用前景[J].中国医学工程,2011,19(5):173-174.
[18] ZHOU Y, WU C, CHANG J. Bioceramics to regulate stem cells and their microenvironment for tissue regeneration. Materials Today. 2019;24: 41-56.
[19] LAM ATL, REUVENY S, OH SK. Human mesenchymal stem cell therapy for cartilage repair: Review on isolation, expansion, and constructs. Stem Cell Res. 2020;44:101738.
[20] 王承云,石磊,夏春,等.人骨髓间充质干细胞分离培养方法的改进[J].中国组织工程研究与临床康复,2011,15(32):5896-5900.
[21] DOMINICI M, LE BLANC K, MUELLER I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8(4):315-317.
[22] 石志红,袁祖贻,张慧峰,等.不同浓度哇巴因对A549细胞增殖及死亡的影响[J].现代肿瘤医学,2011,19(8):1515-1518.
[23] OLIVEIRA TN, POSSIDONIO AC, SOARES CP, et al. The role of Na+/K+-ATPase during chick skeletal myogenesis. PLoS One. 2015;10(3): e0120940.
[24] HUANG T, YU Z, YU Q, et al. Inhibition of osteogenic and adipogenic potential in bone marrow-derived mesenchymal stem cells under osteoporosis. Biochem Biophys Res Commun. 2020;525(4):902-908.
[25] LIU L, IVANOV AV, GABLE ME, et al. Comparative properties of caveolar and noncaveolar preparations of kidney Na+/K+-ATPase. Biochemistry. 2011;50(40):8664-8673.
[26] KUANG Z, BAI J, NI L, et al. Withanolide B promotes osteogenic differentiation of human bone marrow mesenchymal stem cells via ERK1/2 and Wnt/β-catenin signaling pathways. Int Immunopharmacol. 2020;88:106960.