Age-related changes in differentiation of bone marrow mesenchymal stem cells and the activity of Notch signaling pathway

doi:10.19439/j.sjos.2022.02.002

Abstract

Abstract: PURPOSE: To explore the age-related changes in differentiation and proliferation of murine bone marrow mesenchymal stem cells (BMSCs) and the activity of Notch signaling pathway in vitro. METHODS: BMSCs were harvested from the whole bone marrow of young, adult and aged C57BL/6 mice and were evaluated for cell-surface protein expression using flow cytometry. After osteogenic and adipogenic induction, osteogenic and adipogenic differentiation ability of BMSCs was evaluated. Cell viability was analyzed by proliferation and migration assays. The expressions of Notch-related genes were analyzed by real-time PCR. The experimental data were analyzed with SPSS 19.0 software package. RESULTS: Alizarin red S and Oil red O staining results indicated that the osteogenic ability of BMSCs gradually decreased with aging, while the adipogenic ability increased. Cell activity assays showed that the proliferative and migrated capacity did not decline with aging significantly, which suggested that the changes of osteogenic and adipogenic differentiation observed in aged BMSCs were not attributed to cell activity but to differentiation potential. Real-time PCR showed the aged cells exhibited significantly higher Notch signaling expression level than the younger ones (P<0.05). CONCLUSIONS: Decreased bone formation capacity and increased adipogenic differentiation ability were noted in aged BMSCs. At the same time, the activity of Notch signaling pathway shows an aging increase which would propose a new idea to restore the damaged osteogenic differentiation ability of aged BMSCs.

Key words: Aging, Bone marrow mesenchymal stem cells, Osteogenesis, Adipogenesis, Notch

CLC Number:

Q254

TANG Zhao-long, JING Wei. Age-related changes in differentiation of bone marrow mesenchymal stem cells and the activity of Notch signaling pathway[J]. Shanghai Journal of Stomatology, 2022, 31(2): 120-125.

References

[1] Boskey AL, Imbert L.Bone quality changes associated with aging and disease: a review[J]. Ann N Y Acad Sci, 2017, 1410(1): 93-106.
[2] Infante A, Rodríguez CI.Osteogenesis and aging: lessons from mesenchymal stem cells[J]. Stem Cell Res Ther, 2018, 9(1): 244-250.
[3] Mutyaba PL, Belkin NS, Lopas L, et al.Notch signaling in mesenchymal stem cells (MSC) harvested from geriatric mice[J]. J Orthop Trauma, 2014, 28(Suppl 1): S20-S23.
[4] Jing W, Xiong Z, Cai X, et al.Effects of γ-secretase inhibition on the proliferation and vitamin D 3 induced osteogenesis in adipose derived stem cells[J]. Biochem Biophys Res Commun, 2010, 392(3): 442-447.
[5] Huang YK, Yin Y, Gu YZ, et al.Characterization and immunogenicity of bone marrow-derived mesenchymal stem cells under osteoporotic conditions[J]. Sci China Life Sci, 2020, 63(3): 429-442.
[6] Baht GS, Silkstone D, Vi L, et al.Exposure to a youthful circulaton rejuvenates bone repair through modulation of β-catenin[J]. Nat Commun, 2015, 6: 7131.
[7] Chen Q, Shou P, Zheng C, et al.Fate decision of mesenchymal stem cells: adipocytes or osteoblasts?[J]. Cell Death Differ, 2016, 23(7): 1128-1139.
[8] Abdallah BM, Jafari A, Zaher W, et al.Skeletal (stromal) stem cells: an update on intracellular signaling pathways controlling osteoblast differentiation[J]. Bone, 2015, 70: 28-36.
[9] Schultz MB, Sinclair DA.When stem cells grow old: phenotypes and mechanisms of stem cell aging[J]. Development, 2016, 143(1): 3-14.
[10] Friedenstein AJ, Gorskaja JF, Kulagina NN.Fibroblast precursors in normal and irradiated mouse hematopoietic organs[J]. Exp Hematol, 1976, 4(5): 267-274.
[11] Devlin MJ, Rosen CJ.The bone-fat interface: basic and clinical implications of marrow adiposity[J]. Lancet Diabetes Endocrinol, 2015, 3(2): 141-147.
[12] Goodell MA, Rando TA.Stem cells and healthy aging[J]. Science, 2015, 350(6265): 1199-1204.
[13] Qadir AS, Liang S, Wu Z, et al.Senile osteoporosis: the involvement of differentiation and senescence of bone marrow stromal cells[J]. Int J Mol Sci, 2020, 21(1) : 349-356.
[14] Sui BD, Hu CH, Zheng CX, et al.Microenvironmental views on mesenchymal stem cell differentiation in aging[J]. J Dent Res, 2016, 95(12): 1333-1340.
[15] Hojo H, Ohba S, Chung U, et al.Signaling pathways regulating the specification and differentiation of the osteoblast lineage[J]. Regen Ther, 2015, 1: 57-62.
[16] Semenova D, Bogdanova M, Kostina A, et al.Dose-dependent mechanism of Notch action in promoting osteogenic differentiation of mesenchymal stem cells[J]. Cell Tissue Res, 2020, 379(1): 169-179.
[17] Luo Z, Shang X, Zhang H, et al.Notch signaling in osteogenesis, osteoclastogenesis, and angiogenesis[J]. Am J Pathol, 2019, 189(8): 1495-1500.
[18] Wang C, Inzana JA, Mirando AJ, et al.NOTCH signaling in skeletal progenitors is critical for fracture repair[J]. J Clin Invest, 2016, 126(4): 1471-1481.
[19] He Y, Zou L.Notch-1 inhibition reduces proliferation and promotes osteogenic differentiation of bone marrow mesenchymal stem cells[J]. Exp Ther Med, 2019, 18(3): 1884-1890.
[20] Shao J, Zhou Y, Lin J, et al.Notch expressed by osteocytes plays a critical role in mineralisation[J]. J Mol Med (Berl), 2018, 96(3-4): 333-347.