Excessive mechanical stretch‑mediated osteoblasts promote the catabolism and apoptosis of chondrocytes via the Wnt/β‑catenin signaling pathway

Song,Cheng-Xian; Liu,Sheng-Yao; Zhu,Wen-Ting; Xu,Shao-Yong; Ni,Guo-Xin

doi:10.3892/mmr.2021.12232

Excessive mechanical stretch‑mediated osteoblasts promote the catabolism and apoptosis of chondrocytes via the Wnt/β‑catenin signaling pathway

Authors:
- Cheng-Xian Song
- Sheng-Yao Liu
- Wen-Ting Zhu
- Shao-Yong Xu
- Guo-Xin Ni
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Affiliations: Department of Orthopedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Orthopedics, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510260, P.R. China, Department of Pharmacy, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510150, P.R. China
Published online on: June 17, 2021 https://doi.org/10.3892/mmr.2021.12232
Article Number: 593
Copyright: © Song et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Excessive biomechanical loading is considered an important cause of osteoarthritis. Although the mechanical responses of chondrocytes and osteoblasts have been investigated, their communication during mechanical loading and the underlying molecular mechanisms are not yet fully known. The present study investigated the effects of excessive mechanically stretched osteoblasts on the metabolism and apoptosis of chondrocytes, and also assessed the involvement of the Wnt/β‑catenin signaling pathway. In the present study, rat chondrocytes and osteoblasts were subjected to mechanical tensile strain, and an indirect chondrocyte‑osteoblast co‑culture model was established. Reverse transcription‑quantitative PCR and western blotting were performed to determine the expression levels of genes and proteins of interest. An ELISA was performed to investigate the levels of cytokines, including matrix metalloproteinase (MMP) 13, MMP 3, interleukin‑6 (IL‑6) and prostaglandin E2 (PG E2), released from osteoblasts. Flow cytometry was performed to detect the apoptosis of chondrocytes exposed to stretched osteoblast conditioned culture medium. The levels of MMP 13, IL‑6 and PG E2 increased significantly in the supernatants of stretched osteoblasts compared with the un‑stretched group. By contrast, the mRNA expression levels of Collagen 1a and alkaline phosphatase were significantly decreased in osteoblasts subjected to mechanical stretch compared with the un‑stretched group. The mRNA expression level of Collagen 2a was significantly decreased, whereas the expression levels of MMP 13 and a disintegrin and metalloproteinase with thrombospondin‑like motifs 5 were significantly increased in chondrocytes subjected to mechanical stretch compared with the un‑stretched group. In the co‑culture model, the results indicated that excessive mechanically stretched osteoblasts induced the catabolism and apoptosis of chondrocytes, which was partly inhibited by Wnt inhibitor XAV‑939. The results of the present study demonstrated that excessive mechanical stretch led to chondrocyte degradation and inhibited osteoblast osteogenic differentiation; furthermore, excessive mechanically stretched osteoblasts induced the catabolism and apoptosis of chondrocytes via the Wnt/β‑catenin signaling pathway.

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1	Bijlsma JW, Berenbaum F and Lafeber FP: Osteoarthritis: An update with relevance for clinical practice. Lancet. 377:2115–2126. 2011. View Article : Google Scholar : PubMed/NCBI
2	Bowden JL, Hunter DJ, Deveza LA, Duong V, Dziedzic KS, Allen KD, Chan PK and Eyles JP: Core and adjunctive interventions for osteoarthritis: Efficacy and models for implementation. Nat Rev Rheumatol. 16:434–447. 2020. View Article : Google Scholar : PubMed/NCBI
3	Qian J, Liang J, Wang Y and Wang H: Effect of passive motion on articular cartilage in rat osteoarthritis. Exp Ther Med. 8:377–383. 2014. View Article : Google Scholar : PubMed/NCBI
4	Ni GX, Liu SY, Lei L, Li Z, Zhou YZ and Zhan LQ: Intensity-dependent effect of treadmill running on knee articular cartilage in a rat model. BioMed Res Int. 2013:1723922013. View Article : Google Scholar : PubMed/NCBI
5	Goldring MB and Goldring SR: Articular cartilage and subchondral bone in the pathogenesis of osteoarthritis. Ann N Y Acad Sci. 1192:230–237. 2010. View Article : Google Scholar : PubMed/NCBI
6	Goldring SR and Goldring MB: Changes in the osteochondral unit during osteoarthritis: Structure, function and cartilage-bone crosstalk. Nat Rev Rheumatol. 12:632–644. 2016. View Article : Google Scholar : PubMed/NCBI
7	Yao Z, Chen P, Wang S, Deng G, Hu Y, Lin Q, Zhang X and Yu B: Reduced PDGF-AA in subchondral bone leads to articular cartilage degeneration after strenuous running. J Cell Physiol. 234:17946–17958. 2019. View Article : Google Scholar : PubMed/NCBI
8	Sanchez C, Pesesse L, Gabay O, Delcour JP, Msika P, Baudouin C and Henrotin YE: Regulation of subchondral bone osteoblast metabolism by cyclic compression. Arthritis Rheum. 64:1193–1203. 2012. View Article : Google Scholar : PubMed/NCBI
9	Yan YX, Gong YW, Guo Y, Lv Q, Guo C, Zhuang Y, Zhang Y, Li R and Zhang XZ: Mechanical strain regulates osteoblast proliferation through integrin-mediated ERK activation. PLoS One. 7:e357092012. View Article : Google Scholar : PubMed/NCBI
10	Agarwal S, Long P, Gassner R, Piesco NP and Buckley MJ: Cyclic tensile strain suppresses catabolic effects of interleukin-1beta in fibrochondrocytes from the temporomandibular joint. Arthritis Rheum. 44:608–617. 2001. View Article : Google Scholar : PubMed/NCBI
11	Zhu G, Qian Y, Wu W and Li R: Negative effects of high mechanical tensile strain stimulation on chondrocyte injury in vitro. Biochem Biophys Res Commun. 510:48–52. 2019. View Article : Google Scholar : PubMed/NCBI
12	Takebe K, Nishiyama T, Hayashi S, Hashimoto S, Fujishiro T, Kanzaki N, Kawakita K, Iwasa K, Kuroda R and Kurosaka M: Regulation of p38 MAPK phosphorylation inhibits chondrocyte apoptosis in response to heat stress or mechanical stress. Int J Mol Med. 27:329–335. 2011.PubMed/NCBI
13	Kaufmann SH, Lee SH, Meng XW, Loegering DA, Kottke TJ, Henzing AJ, Ruchaud S, Samejima K and Earnshaw WC: Apoptosis associated caspase activation assays. Methods. 44:262–272. 2008. View Article : Google Scholar : PubMed/NCBI
14	Iannone F and Lapadula G: Phenotype of chondrocytes in osteoarthritis. Biorheology. 45:411–413. 2008. View Article : Google Scholar : PubMed/NCBI
15	Laowanitwattana T, Aungsuchawan S, Narakornsak S, Markmee R, Tancharoen W, Keawdee J, Boonma N, Tasuya W, Peerapapong L, Pangjaidee N, et al: Osteoblastic differentiation potential of human amniotic fluid-derived mesenchymal stem cells in different culture conditions. Acta Histochem. 120:701–712. 2018. View Article : Google Scholar : PubMed/NCBI
16	Lefebvre V and Dvir-Ginzberg M: SOX9 and the many facets of its regulation in the chondrocyte lineage. Connect Tissue Res. 58:2–14. 2017. View Article : Google Scholar : PubMed/NCBI
17	Bleuel J, Zaucke F, Brüggemann GP and Niehoff A: Effects of cyclic tensile strain on chondrocyte metabolism: A systematic review. PLoS One. 10:e01198162015. View Article : Google Scholar : PubMed/NCBI
18	Tetlow LC, Adlam DJ and Woolley DE: Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage: Associations with degenerative changes. Arthritis Rheum. 44:585–594. 2001. View Article : Google Scholar : PubMed/NCBI
19	Sakao K, Takahashi KA, Mazda O, Arai Y, Tonomura H, Inoue A, Saito M, Fujioka M, Takamiya H, Imanishi J, et al: Enhanced expression of interleukin-6, matrix metalloproteinase-13, and receptor activator of NF-kappaB ligand in cells derived from osteoarthritic subchondral bone. J Orthop Sci. 13:202–210. 2008. View Article : Google Scholar : PubMed/NCBI
20	Verma P and Dalal K: ADAMTS-4 and ADAMTS-5: Key enzymes in osteoarthritis. J Cell Biochem. 112:3507–3514. 2011. View Article : Google Scholar : PubMed/NCBI
21	Ruan G, Xu J, Wang K, Wu J, Zhu Q, Ren J, Bian F, Chang B, Bai X, Han W, et al: Associations between knee structural measures, circulating inflammatory factors and MMP13 in patients with knee osteoarthritis. Osteoarthritis Cartilage. 26:1063–1069. 2018. View Article : Google Scholar : PubMed/NCBI
22	Nusse R: Wnt signaling in disease and in development. Cell Res. 15:28–32. 2005. View Article : Google Scholar : PubMed/NCBI
23	Oh H, Chun CH and Chun JS: Dkk-1 expression in chondrocytes inhibits experimental osteoarthritic cartilage destruction in mice. Arthritis Rheum. 64:2568–2578. 2012. View Article : Google Scholar : PubMed/NCBI
24	Miclea RL, Siebelt M, Finos L, Goeman JJ, Löwik CW, Oostdijk W, Weinans H, Wit JM, Robanus-Maandag EC and Karperien M: Inhibition of Gsk3β in cartilage induces osteoarthritic features through activation of the canonical Wnt signaling pathway. Osteoarthritis Cartilage. 19:1363–1372. 2011. View Article : Google Scholar : PubMed/NCBI
25	Chen M, Zhu M, Awad H, Li TF, Sheu TJ, Boyce BF, Chen D and O'Keefe RJ: Inhibition of beta-catenin signaling causes defects in postnatal cartilage development. J Cell Sci. 121:1455–1465. 2008. View Article : Google Scholar : PubMed/NCBI
26	Gordon MD and Nusse R: Wnt signaling: Multiple pathways, multiple receptors, and multiple transcription factors. J Biol Chem. 281:22429–22433. 2006. View Article : Google Scholar : PubMed/NCBI
27	Niu Q, Li F, Zhang L, Xu X, Liu Y, Gao J and Feng X: Role of the Wnt/β-catenin signaling pathway in the response of chondrocytes to mechanical loading. Int J Mol Med. 37:755–762. 2016. View Article : Google Scholar : PubMed/NCBI
28	Liedert A, Wagner L, Seefried L, Ebert R, Jakob F and Ignatius A: Estrogen receptor and Wnt signaling interact to regulate early gene expression in response to mechanical strain in osteoblastic cells. Biochem Biophys Res Commun. 394:755–759. 2010. View Article : Google Scholar : PubMed/NCBI
29	Yan YB, Li JM, Xiao E, An JG, Gan YH and Zhang Y: A pilot trial on the molecular pathophysiology of traumatic temporomandibular joint bony ankylosis in a sheep model. Part I: Expression of Wnt signaling. J Craniomaxillofac Surg. 42:e15–e22. 2014. View Article : Google Scholar
30	Chen X, Guo J, Yuan Y, Sun Z, Chen B, Tong X, Zhang L, Shen C and Zou J: Cyclic compression stimulates osteoblast differentiation via activation of the Wnt/β-catenin signaling pathway. Mol Med Rep. 15:2890–2896. 2017. View Article : Google Scholar : PubMed/NCBI
31	Fushiki R, Mayahara K, Ogawa M, Takahashi Y, Karasawa Y, Tsurumachi N, Tamura T and Shimizu N: High-magnitude mechanical strain inhibits the differentiation of bone-forming rat calvarial progenitor cells. Connect Tissue Res. 56:336–341. 2015. View Article : Google Scholar : PubMed/NCBI
32	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
33	Zhang RK, Li GW, Zeng C, Lin CX, Huang LS, Huang GX, Zhao C, Feng SY and Fang H: Mechanical stress contributes to osteoarthritis development through the activation of transforming growth factor beta 1 (TGF-β1). Bone Joint Res. 7:587–594. 2018. View Article : Google Scholar : PubMed/NCBI
34	Findlay DM and Atkins GJ: Osteoblast-chondrocyte interactions in osteoarthritis. Curr Osteoporos Rep. 12:127–134. 2014. View Article : Google Scholar : PubMed/NCBI
35	Yuan XL, Meng HY, Wang YC, Peng J, Guo QY, Wang AY and Lu SB: Bone-cartilage interface crosstalk in osteoarthritis: Potential pathways and future therapeutic strategies. Osteoarthritis Cartilage. 22:1077–1089. 2014. View Article : Google Scholar : PubMed/NCBI
36	Li Z, Liu SY, Xu L, Xu SY and Ni GX: Effects of treadmill running with different intensity on rat subchondral bone. Sci Rep. 7:19772017. View Article : Google Scholar : PubMed/NCBI
37	Liu SY, Li Z, Xu SY, Xu L, Yang M and Ni GX: Intensity dependent effect of treadmill running on differentiation of rat bone marrow stromal cells. Mol Med Rep. 17:7746–7756. 2018.PubMed/NCBI
38	Tang L, Lin Z and Li YM: Effects of different magnitudes of mechanical strain on Osteoblasts in vitro. Biochem Biophys Res Commun. 344:122–128. 2006. View Article : Google Scholar : PubMed/NCBI
39	Lin YY, Tanaka N, Ohkuma S, Iwabuchi Y, Tanne Y, Kamiya T, Kunimatsu R, Huang YC, Yoshioka M, Mitsuyoshi T, et al: Applying an excessive mechanical stress alters the effect of subchondral osteoblasts on chondrocytes in a co-culture system. Eur J Oral Sci. 118:151–158. 2010. View Article : Google Scholar : PubMed/NCBI
40	Zhang WM, Liu Y, Li TT, Piao CM, Liu O, Liu JL, Qi YF, Jia LX and Du J: Sustained activation of ADP/P2ry12 signaling induces SMC senescence contributing to thoracic aortic aneurysm/dissection. J Mol Cell Cardiol. 99:76–86. 2016. View Article : Google Scholar : PubMed/NCBI
41	Hu W, Chen Y, Dou C and Dong S: Microenvironment in subchondral bone: Predominant regulator for the treatment of osteoarthritis. Ann Rheum Dis. 80:413–422. 2020. View Article : Google Scholar : PubMed/NCBI
42	Saltzman BM and Riboh JC: Subchondral bone and the osteochondral unit: Basic science and clinical implications in sports medicine. Sports Health. 10:412–418. 2018. View Article : Google Scholar : PubMed/NCBI
43	Jiang J, Nicoll SB and Lu HH: Co-culture of osteoblasts and chondrocytes modulates cellular differentiation in vitro. Biochem Biophys Res Commun. 338:762–770. 2005. View Article : Google Scholar : PubMed/NCBI
44	Sanchez C, Deberg MA, Piccardi N, Msika P, Reginster JY and Henrotin YE: Subchondral bone osteoblasts induce phenotypic changes in human osteoarthritic chondrocytes. Osteoarthritis Cartilage. 13:988–997. 2005. View Article : Google Scholar : PubMed/NCBI
45	Priam S, Bougault C, Houard X, Gosset M, Salvat C, Berenbaum F and Jacques C: Identification of soluble 14-3-3є as a novel subchondral bone mediator involved in cartilage degradation in osteoarthritis. Arthritis Rheum. 65:1831–1842. 2013. View Article : Google Scholar : PubMed/NCBI
46	Sanchez C, Deberg MA, Piccardi N, Msika P, Reginster JY and Henrotin YE: Osteoblasts from the sclerotic subchondral bone downregulate aggrecan but upregulate metalloproteinases expression by chondrocytes. This effect is mimicked by interleukin-6, −1beta and oncostatin M pre-treated non-sclerotic osteoblasts. Osteoarthritis Cartilage. 13:979–987. 2005. View Article : Google Scholar : PubMed/NCBI
47	Sasaki K, Takagi M, Konttinen YT, Sasaki A, Tamaki Y, Ogino T, Santavirta S and Salo J: Upregulation of matrix metalloproteinase (MMP)-1 and its activator MMP-3 of human osteoblast by uniaxial cyclic stimulation. J Biomed Mater Res B Appl Biomater. 80:491–498. 2007. View Article : Google Scholar : PubMed/NCBI
48	Fermor B, Gundle R, Evans M, Emerton M, Pocock A and Murray D: Primary human osteoblast proliferation and prostaglandin E2 release in response to mechanical strain in vitro. Bone. 22:637–643. 1998. View Article : Google Scholar : PubMed/NCBI
49	Akhtar N, Khan NM, Ashruf OS and Haqqi TM: Inhibition of cartilage degradation and suppression of PGE2 and MMPs expression by pomegranate fruit extract in a model of posttraumatic osteoarthritis. Nutrition. 33:1–13. 2017. View Article : Google Scholar : PubMed/NCBI
50	Weng LH, Wang CJ, Ko JY, Sun YC and Wang FS: Control of Dkk-1 ameliorates chondrocyte apoptosis, cartilage destruction, and subchondral bone deterioration in osteoarthritic knees. Arthritis Rheum. 62:1393–1402. 2010. View Article : Google Scholar : PubMed/NCBI
51	Prasadam I, Friis T, Shi W, van Gennip S, Crawford R and Xiao Y: Osteoarthritic cartilage chondrocytes alter subchondral bone osteoblast differentiation via MAPK signalling pathway involving ERK1/2. Bone. 46:226–235. 2010. View Article : Google Scholar : PubMed/NCBI
52	Kimura H, Fumoto K, Shojima K, Nojima S, Osugi Y, Tomihara H, Eguchi H, Shintani Y, Endo H, Inoue M, et al: CKAP4 is a Dickkopf1 receptor and is involved in tumor progression. J Clin Invest. 126:2689–2705. 2016. View Article : Google Scholar : PubMed/NCBI