Effect of a disintegrin and metalloproteases 10 on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells induced by steroid-induced osteonecrosis of femoral head

doi:10.12307/2023.369

Abstract

Abstract: BACKGROUND: Many studies have found that the pathological progression of steroid-induced osteonecrosis of the femoral head is related to abnormal proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, in which a disintegrin and metalloproteases 10 (ADAM10) is highly involved, but the specific mechanism remains unclear.
OBJECTIVE: To investigate the effects of ADAM10 on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells derived from steroid-induced osteonecrosis of the femoral head patients and its possible mechanism.
METHODS: Bone marrow was harvested from proximal femur in 23 patients with steroid-induced osteonecrosis of the femoral head and 17 patients with femoral neck fracture under sterile conditions. Bone marrow mesenchymal stem cells from steroid-induced osteonecrosis of the femoral head (S-BMSCs) and femoral neck fracture (F-BMSCs) were isolated by density gradient centrifugation, and cultured to the third generation in vitro. The proliferation activity of cells in the two groups was detected by CCK-8 assay. Alizarin red staining and alkaline phosphatase staining were used to observe the osteogenic differentiation ability of cells in the two groups. qRT-PCR and western blot assay were used to detect the levels of ADAM10 mRNA and protein in cells of the two groups. S-BMSCs were infected with ADAM10 overexpression lentivirus vector, and then treated with Notch signaling pathway inhibitor DAPT. The proliferation activity of these cells was detected by CCK-8 assay. Alizarin red staining and alkaline phosphatase staining were used to observe the osteogenic differentiation of cells. qRT-PCR was used to detect the expression levels of osteogenic markers alkaline phosphatase, Runt-related transcription factor 2, and osteocalcin mRNA. The expression levels of Notch signaling pathway related proteins Notch1, NICD, and Hes1 were detected by western blot assay.
RESULTS AND CONCLUSION: (1) The proliferation activity, osteogenic differentiation ability and the expression level of ADAM10 in S-BMSCs were significantly lower than those in F-BMSCs (P < 0.01). (2) After ADAM10 overexpression, the proliferation activity, osteogenic differentiation ability, the expression levels of alkaline phosphatase, Runt-related transcription factor 2, and osteocalcin mRNA and Notch1, NICD and Hes1 protein of S-BMSCs were significantly increased (P < 0.05). However, combined intervention with DAPT significantly inhibited the promoting effects of ADAM10 overexpression on proliferation activity and osteogenic differentiation ability of S-BMSCs, and inhibited the activation of Notch signaling pathway. (3) The results show that ADAM10 can be underexpressed in S-BMSCs, and its overexpression can enhance the proliferation and osteogenic differentiation ability of S-BMSCs, and its mechanism may be related to the activation of Notch signaling pathway.

Key words: steroid-induced osteonecrosis of femoral head, bone marrow mesenchymal stem cell, a disintegrin and metalloproteases 10, Notch signaling pathway, osteogenic differentiation

CLC Number:

Fan Jun, Wu Chenhuan, Cheng Zhonghua. Effect of a disintegrin and metalloproteases 10 on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells induced by steroid-induced osteonecrosis of femoral head[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(10): 1507-1513.

Figures/Tables 9

References

[1] WU X, SUN W, TAN M. Noncoding RNAs in Steroid-Induced Osteonecrosis of the Femoral Head. Biomed Res Int. 2019;2019: 8140595.
[2] HAN L, WANG B, WANG R, et al. The shift in the balance between osteoblastogenesis and adipogenesis of mesenchymal stem cells mediated by glucocorticoid receptor. Stem Cell Res Ther. 2019;10(1): 377.
[3] WANG SH, GOU GH, WU CC, et al. Increased COUP-TFII Expression Mediates the Differentiation Imbalance of Bone Marrow-Derived Mesenchymal Stem Cells in Femoral Head Osteonecrosis. Biomed Res Int. 2019;2019:9262430.
[4] WANG T, TENG S, ZHANG Y, et al. Role of mesenchymal stem cells on differentiation in steroid-induced avascular necrosis of the femoral head. Exp Ther Med. 2017;13(2):669-675.
[5] HAO C, YANG S, XU W, et al. MiR-708 promotes steroid-induced osteonecrosis of femoral head, suppresses osteogenic differentiation by targeting SMAD3. Sci Rep. 2016;6:22599.
[6] ZIOUTI F, EBERT R, RUMMLER M, et al. NOTCH Signaling Is Activated through Mechanical Strain in Human Bone Marrow-Derived Mesenchymal Stromal Cells. Stem Cells Int. 2019;2019:5150634.
[7] VAN TETERING G, VAN DIEST P, VERLAAN I, et al. Metalloprotease ADAM10 is required for Notch1 site 2 cleavage. J Biol Chem. 2009; 284(45):31018-31027.
[8] 朱永翠, 翟蕾, 闫亚姿, 等.过表达ADAM10通过调控Notch信号通路对牙周膜干细胞成骨分化的影响[J].口腔医学研究,2021, 37(5):468-473.
[9] 中国医师协会骨科医师分会显微修复工作委员会. 成人股骨头坏死临床诊疗指南(2016)[J]. 中华骨科杂志,2016,36(15):945-954.
[10] KOO KH, KIM R, KIM YS, et al. Risk period for developing osteonecrosis of the femoral head in patients on steroid treatment. Clin Rheumatol. 2002;21(4):299-303.
[11] LV Z, MUHEREMU A, BAI X, et al. PTH(1‑34) activates the migration and adhesion of BMSCs through the rictor/mTORC2 pathway. Int J Mol Med. 2020;46(6):2089-2101.
[12] CHENG S, LIU X, GONG F, et al. Dexamethasone promotes the endoplasmic reticulum stress response of bone marrow mesenchymal stem cells by activating the PERK-Nrf2 signaling pathway. Pharmacol Res Perspect. 2021;9(3):e00791.
[13] ZHANG P, TAO F, LI Q, et al. 5-Azacytidine and trichostatin A enhance the osteogenic differentiation of bone marrow mesenchymal stem cells isolated from steroid-induced avascular necrosis of the femoral head in rabbit. J Biosci. 2019;44(4):87.
[14] SMITH TM JR, THARAKAN A, MARTIN RK. Targeting ADAM10 in Cancer and Autoimmunity. Front Immunol. 2020;11:499.
[15] YUAN Q, YU H, CHEN J, et al. ADAM10 promotes cell growth, migration, and invasion in osteosarcoma via regulating E-cadherin/β-catenin signaling pathway and is regulated by miR-122-5p. Cancer Cell Int. 2020;20:99.
[16] ARAYA HF, SEPULVEDA H, LIZAMA CO, et al. Expression of the ectodomain-releasing protease ADAM17 is directly regulated by the osteosarcoma and bone-related transcription factor RUNX2. J Cell Biochem. 2018;119(10):8204-8219.
[17] 谢映春, 涂小林. 体外激活骨细胞Notch信号对骨髓基质细胞成骨分化的影响[J]. 第三军医大学学报,2020,42(9):891-898.
[18] ZHANG H, LIN J, CHEN J, et al. Effects of Notch/p38MAPK signaling pathway on articular cartilage defect recovery by BMSCs tissue based on the rabbit articular cartilage defect models. Saudi J Biol Sci. 2020; 27(3):859-864.
[19] WANG G, YAN J, ZHANG H, et al. Transient activation of notch signaling enhances endogenous stromal cell expansion and subsequent bone defect repair. J Orthop Translat. 2021;31:26-32.
[20] LINDSEY RC, XING W, POURTEYMOOR S, et al. Novel Role for Claudin-11 in the Regulation of Osteoblasts via Modulation of ADAM10-Mediated Notch Signaling. J Bone Miner Res. 2019;34(10):1910-1922.