Cyclic compression stimulates osteoblast differentiation via activation of the Wnt/β-catenin signaling pathway

Chen,Xi; Guo,Jianmin; Yuan,Yu; Sun,Zhongguang; Chen,Binglin; Tong,Xiaoyang; Zhang,Lingli; Shen,Chao; Zou,Jun

doi:10.3892/mmr.2017.6327

Cyclic compression stimulates osteoblast differentiation via activation of the Wnt/β-catenin signaling pathway

Authors:
- Xi Chen
- Jianmin Guo
- Yu Yuan
- Zhongguang Sun
- Binglin Chen
- Xiaoyang Tong
- Lingli Zhang
- Chao Shen
- Jun Zou
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Affiliations: School of Sports Science, Wenzhou Medical University, Wenzhou, Zhejiang 325035, P.R. China, School of Kinesiology, Shanghai University of Sport, Shanghai 200438, P.R. China, Development and Planning Department, Shanghai University of Sport, Shanghai 200438, P.R. China
Published online on: March 15, 2017 https://doi.org/10.3892/mmr.2017.6327
Pages: 2890-2896

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Abstract

It is widely accepted that mechanical stress is an important factor in bone associated cell differentiation, including that of mesenchymal stem cells, osteoblasts and osteocytes. The present study aimed to determine the effect of mechanical cyclic compressive load on osteoblast differentiation, and whether this was associated with activation of the wingless‑type (Wnt)/β-catenin signaling pathway. Using a 3D scaffold model, MC3T3‑E1 cells were exposed to cyclic compressive loading via the Flexcell‑5000C™ Compression system. Sinusoidal wave magnitudes of 0.33, 0.5 and 1 MPa were applied for 4, 6 and 8 h, at 1 Hz frequency. Expression levels of genes associated with osteoblast differentiation were enhanced following compression, including alkaline phosphatase, osteocalcin, runt‑related transcription factor 2 and osterix. Optimal compression was observed using a magnitude of 0.5 MPa for 6 h, whereas a magnitude of 1 MPa had no effect on osteoblast differentiation, and had a negative effect when applied for prolonged time periods. Compressive loading additionally enhanced the mRNA expression levels of the Wnt/β‑catenin signaling pathway component, low density lipoprotein receptor‑related protein 5, and the protein expression levels of Wnt1, disheveled segment polarity protein‑2 (DVL2) and β-catenin. By contrast, mRNA expression levels of sclerostin and the inactive form of β-catenin (phosphorylated at Ser33/37/Thr41) were reduced following compressive loading. Following compressive loading of cells, dickkopf-related protein 1 (DKK‑1), an inhibitor of the Wnt signaling pathway, increased protein expression levels of the inactive form of the Wnt‑associated protein, phosphorylated‑β‑catenin, compared with compression alone. However, DVL2 and Wnt1 protein expression levels were unaffected, suggesting that the loading‑induced activation of Wnt/β‑catenin signaling decreased however, it was not prevented by DKK‑1 treatment. In conclusion, the present study demonstrated that cyclic compressive load promoted osteoblast differentiation and may be dependent on the Wnt/β-catenin signaling pathway in regard to magnitude and duration.

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1	NIH Consensus Development Panel on Osteoporosis Prevention, Diagnosis, and Therapy: Osteoporosis prevention, diagnosis, and therapy. JAMA. 285:785–795. 2001. View Article : Google Scholar : PubMed/NCBI
2	Bliuc D, Alarkawi D, Nguyen TV, Eisman JA and Center JR: Risk of subsequent fractures and mortality in elderly women and men with fragility fractures with and without osteoporotic bone density: The dubbo osteoporosis epidemiology study. J Bone Miner Res. 30:637–646. 2015. View Article : Google Scholar : PubMed/NCBI
3	Zaidi M: Skeletal remodeling in health and disease. Nature Med. 13:791–801. 2007. View Article : Google Scholar : PubMed/NCBI
4	Lang T, LeBlanc A, Evans H, Lu Y, Genant H and Yu A: Cortical and trabecular bone mineral loss from the spine and hip in long-duration spaceflight. J Bone Miner Res. 19:1006–1012. 2004. View Article : Google Scholar : PubMed/NCBI
5	Kim CH, Kim KH and Jacobs CR: Effects of high frequency loading on RANKL and OPG mRNA expression in ST-2 murine stromal cells. BMC Musculoskelet Disord. 10:1092009. View Article : Google Scholar : PubMed/NCBI
6	Ponik SM, Triplett JW and Pavalko FM: Osteoblasts and osteocytes respond differently to oscillatory and unidirectional fluid flow profiles. J Cell Biochem. 100:794–807. 2007. View Article : Google Scholar : PubMed/NCBI
7	Carinci F, Pezzetti F, Spina AM, Palmieri A, Carls F, Laino G, De Rosa A, Farina E, Illiano F, Stabellini G, et al: An in vitro model for dissecting distraction osteogenesis. J Craniofac Surg. 16:71–79. 2005. View Article : Google Scholar : PubMed/NCBI
8	Sittichockechaiwut A, Scutt AM, Ryan AJ, Bonewald LF and Reilly GC: Use of rapidly mineralising osteoblasts and short periods of mechanical loading to accelerate matrix maturation in 3D scaffolds. Bone. 44:822–829. 2009. View Article : Google Scholar : PubMed/NCBI
9	Zhong Z, Zeng XL, Ni JH and Huang XF: Comparison of the biological response of osteoblasts after tension and compression. Eur J Orthod. 35:59–65. 2013. View Article : Google Scholar : PubMed/NCBI
10	Thompson WR, Rubin CT and Rubin J: Mechanical regulation of signaling pathways in bone. Gene. 503:179–193. 2012. View Article : Google Scholar : PubMed/NCBI
11	Robling AG, Niziolek PJ, Baldridge LA, Condon KW, Allen MR, Alam I, Mantila SM, Gluhak-Heinrich J, Bellido TM, Harris SE and Turner CH: Mechanical stimulation of bone in vivo reduces osteocyte expression of Sost/sclerostin. J Biol Chem. 283:5866–5875. 2008. View Article : Google Scholar : PubMed/NCBI
12	Wozniak M, Fausto A, Carron CP, Meyer DM and Hruska KA: Mechanically strained cells of the osteoblast lineage organize their extracellular matrix through unique sites of alphavbeta3-integrin expression. J Bone Miner Res. 15:1731–1745. 2000. View Article : Google Scholar : PubMed/NCBI
13	Day TF, Guo X, Garrett-Beal L and Yang Y: Wnt/beta-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev Cell. 8:739–750. 2005. View Article : Google Scholar : PubMed/NCBI
14	Bennett CN, Longo KA, Wright WS, Suva LJ, Lane TF, Hankenson KD and MacDougald OA: Regulation of osteoblastogenesis and bone mass by Wnt10b. Proc Natl Acad Sci USA. 102:3324–3329. 2005. View Article : Google Scholar : PubMed/NCBI
15	Canalis E: Wnt signalling in osteoporosis: Mechanisms and novel therapeutic approaches. Nat Rev Endocrinol. 9:575–583. 2013. View Article : Google Scholar : PubMed/NCBI
16	Kato M, Patel MS, Levasseur R, Lobov I, Chang BH, Glass DA II, Hartmann C, Li L, Hwang TH, Brayton CF, et al: Cbfa1-independent decrease in osteoblast proliferation, osteopenia and persistent embryonic eye vascularization in mice deficient in Lrp5, a Wnt coreceptor. J Cell Biol. 157:303–314. 2002. View Article : Google Scholar : PubMed/NCBI
17	Krishnan V, Bryant HU and Macdougald OA: Regulation of bone mass by Wnt signaling. J Clin Invest. 116:1202–1209. 2006. View Article : Google Scholar : PubMed/NCBI
18	Gong Y, Slee RB, Fukai N, Rawadi G, Roman-Roman S, Reginato AM, Wang H, Cundy T, Glorieux FH, Lev D, et al: LDL receptor-related protein 5 (LRP5) affects bone accrual and eye development. Cell. 107:513–523. 2001. View Article : Google Scholar : PubMed/NCBI
19	Cui Y, Niziolek PJ, MacDonald BT, Zylstra CR, Alenina N, Robinson DR, Zhong Z, Matthes S, Jacobsen CM, Conlon RA, et al: Lrp5 functions in bone to regulate bone mass. Nat Med. 17:684–691. 2011. View Article : Google Scholar : PubMed/NCBI
20	Lian JB, Stein GS, Javed A, van Wijnen AJ, Stein JL, Montecino M, Hassan MQ, Gaur T, Lengner CJ and Young DW: Networks and hubs for the transcriptional control of osteoblastogenesis. Rev Endocr Metab Disord. 7:1–16. 2006. View Article : Google Scholar : PubMed/NCBI
21	Javed A, Bae JS, Afzal F, Gutierrez S, Pratap J, Zaidi SK, Lou Y, van Wijnen AJ, Stein JL, Stein GS and Lian JB: Structural coupling of Smad and Runx2 for execution of the BMP2 osteogenic signal. J Biol Chem. 283:8412–8422. 2008. View Article : Google Scholar : PubMed/NCBI
22	Sawakami K, Robling AG, Ai M, Pitner ND, Liu D, Warden SJ, Li J, Maye P, Rowe DW, Duncan RL, et al: The Wnt co-receptor LRP5 is essential for skeletal mechanotransduction but not for the anabolic bone response to parathyroid hormone treatment. J Biol Chem. 281:23698–23711. 2006. View Article : Google Scholar : PubMed/NCBI
23	Sen B, Guilluy C, Xie Z, Case N, Styner M, Thomas J, Oguz I, Rubin C, Burridge K and Rubin J: Mechanically induced focal adhesion assembly amplifies anti-adipogenic pathways in mesenchymal stem cells. Stem cells. 29:1829–1836. 2011. View Article : Google Scholar : PubMed/NCBI
24	Tu X, Rhee Y, Condon KW, Bivi N, Allen MR, Dwyer D, Stolina M, Turner CH, Robling AG, Plotkin LI and Bellido T: Sost downregulation and local Wnt signaling are required for the osteogenic response to mechanical loading. Bone. 50:209–217. 2012. View Article : Google Scholar : PubMed/NCBI
25	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
26	Koike M, Shimokawa H, Kanno Z, Ohya K and Soma K: Effects of mechanical strain on proliferation and differentiation of bone marrow stromal cell line ST2. J Bone Miner Metab. 23:219–225. 2005. View Article : Google Scholar : PubMed/NCBI
27	Jacobs C, Grimm S, Ziebart T, Walter C and Wehrbein H: Osteogenic differentiation of periodontal fibroblasts is dependent on the strength of mechanical strain. Arch Oral Biol. 58:896–904. 2013. View Article : Google Scholar : PubMed/NCBI
28	Sanchez C, Gabay O, Salvat C, Henrotin YE and Berenbaum F: Mechanical loading highly increases IL-6 production and decreases OPG expression by osteoblasts. Osteoarthritis Cartilage. 17:473–481. 2009. View Article : Google Scholar : PubMed/NCBI
29	Fermor B, Weinberg JB, Pisetsky DS, Misukonis MA, Banes AJ and Guilak F: The effects of static and intermittent compression on nitric oxide production in articular cartilage explants. J Orthop Res. 19:729–737. 2001. View Article : Google Scholar : PubMed/NCBI
30	Xue Y, Yan Y, Gong H, Fang B, Zhou Y, Ding Z, Yin P, Zhang G, Ye Y, Yang C, et al: Insulin-like growth factor binding protein 4 enhances cardiomyocytes induction in murine-induced pluripotent stem cells. J Cell Biochem. 115:1495–1504. 2014. View Article : Google Scholar : PubMed/NCBI
31	Lviak 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
32	Dumas V, Perrier A, Malaval L, Laroche N, Guignandon A, Vico L and Rattner A: The effect of dual frequency cyclic compression on matrix deposition by osteoblast-like cells grown in 3D scaffolds and on modulation of VEGF variant expression. Biomaterials. 30:3279–3288. 2009. View Article : Google Scholar : PubMed/NCBI
33	David V, Guignandon A, Martin A, Malaval L, Lafage-Proust MH, Rattner A, Mann V, Noble B, Jones DB and Vico L: Ex Vivo bone formation in bovine trabecular bone cultured in a dynamic 3D bioreactor is enhanced by compressive mechanical strain. Tissue Eng Part A. 14:117–126. 2008. View Article : Google Scholar : PubMed/NCBI
34	Peptan AI, Lopez A, Kopher RA and Mao JJ: Responses of intramembranous bone and sutures upon in vivo cyclic tensile and compressive loading. Bone. 42:432–438. 2008. View Article : Google Scholar : PubMed/NCBI
35	Goel S, Chin EN, Fakhraldeen SA, Berry SM, Beebe DJ and Alexander CM: Both LRP5 and LRP6 receptors are required to respond to physiological Wnt ligands in mammary epithelial cells and fibroblasts. J Biol Chem. 287:16454–16466. 2012. View Article : Google Scholar : PubMed/NCBI
36	Colaianni G, Cuscito C, Mongelli T, Pignataro P, Tamma R, Oranger A, Colucci S and Grano M: Cellular Mechanisms of Bone Regeneration: Role of Wnt-1 in Bone-Muscle Interaction during Physical Activity39. J Biol Regul Homeost Agents. 29:(4 Suppl). S39–S45. 2015.
37	MacDonald BT, Tamai K and He X: Wnt/beta-catenin signaling: Components, mechanisms, and diseases. Dev Cell. 17:9–26. 2009. View Article : Google Scholar : PubMed/NCBI
38	Baron R and Kneissel M: WNT signaling in bone homeostasis and disease: From human mutations to treatments. Nat Med. 19:179–192. 2013. View Article : Google Scholar : PubMed/NCBI
39	Robinson JA, Chatterjee-Kishore M, Yaworsky PJ, Cullen DM, Zhao W, Li C, Kharode Y, Sauter L, Babij P, Brown EL, et al: Wnt/beta-catenin signaling is a normal physiological response to mechanical loading in bone. J Biol Chem. 281:31720–31728. 2006. View Article : Google Scholar : PubMed/NCBI
40	He XC, Zhang J, Tong WG, Tawfik O, Ross J, Scoville DH, Tian Q, Zeng X, He X, Wiedemann LM, et al: BMP signaling inhibits intestinal stem cell self-renewal through suppression of Wnt-beta-catenin signaling. Nat Genet. 36:1117–1121. 2004. View Article : Google Scholar : PubMed/NCBI
41	Case N, Ma M, Sen B, Xie Z, Gross TS and Rubin J: Beta-catenin levels influence rapid mechanical responses in osteoblasts. J Biol Chem. 283:29196–29205. 2008. View Article : Google Scholar : PubMed/NCBI