Degree of endplate chondrocyte degeneration in different tension regions during mechanical stimulation

Xu,Yongming; Xu,Shujuan; Gao,Zhi; Xiao,Liang; Xiao,Fei; Xu,Hongguang; Zhang,Xiaoling

doi:10.3892/mmr.2018.8435

Degree of endplate chondrocyte degeneration in different tension regions during mechanical stimulation

Authors:
- Yongming Xu
- Shujuan Xu
- Zhi Gao
- Liang Xiao
- Fei Xiao
- Hongguang Xu
- Xiaoling Zhang
View Affiliations

Affiliations: Department of Orthopedic Surgery, Yijishan Hospital, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui 241001, P.R. China, Department of Surgery, Yijishan Hospital, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui 241001, P.R. China, Department of Orthopedic Surgery, Xinhua Hospital Afﬁliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China
Published online on: January 16, 2018 https://doi.org/10.3892/mmr.2018.8435
Pages: 4415-4421
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of this study was to explore the degree of degeneration of endplate chondrocytes in different tension regions induced by intermittent cyclic mechanical tension (ICMT) in vitro. Rat endplate chondrocytes were harvested and treated with 10% ICMT for 8 h/day with a frequency of 0.5 Hz. A cartilage degeneration model was induced using an FX‑5000T cell strain‑loading system. The experiment was divided into the central region and the peripheral region, according to the contact area between the loading post and the six‑well ﬂexible silicone rubber BioFlex plates. Toluidine blue and phalloidin staining were used to observe the morphological changes of cells following mechanical stimulation. Apoptosis was detected by flow cytometry and the mRNA and protein expression levels of collagen type II α1, aggrecan, SRY‑box 9 and matrix metalloproteinase 13 were detected by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blotting, respectively. Endplate chondrocytes exhibited degenerative alterations under mechanical conditions of 10% ICMT and 0.5 Hz at 8 h/day. Toluidine blue and phalloidin staining demonstrated that the cells in the peripheral region were more slender compared with cells in the central region, but RT‑qPCR and western blotting results demonstrated that the degree of cell degeneration between the two groups was not statistically differences. So that cell morphological alteration does not imply that cells have undergone degeneration.

View Figures

View References

1	Manek NJ and MacGregor AJ: Epidemiology of back disorders: Prevalence, risk factors, and prognosis. Curr Opin Rheumatol. 17:134–140. 2005.PubMed/NCBI
2	Rodriguez AG, Rodriguez-Soto AE, Burghardt AJ, Berven S, Majumdar S and Lotz JC: Morphology of the human vertebral endplate. J Orthop Res. 30:280–287. 2012. View Article : Google Scholar : PubMed/NCBI
3	Walter BA, Korecki CL, Purmessur D, Roughley PJ, Michalek AJ and Iatridis JC: Complex loading affects intervertebral disc mechanics and biology. Osteoarthritis Cartilage. 19:1011–1018. 2011. View Article : Google Scholar : PubMed/NCBI
4	Vergroesen PP, Kingma I, Emanuel KS, Hoogendoorn RJ, Welting TJ, van Royen BJ, van Dieën JH and Smit TH: Mechanics and biology in intervertebral disc degeneration: A vicious circle. Osteoarthritis Cartilage. 23:1057–1070. 2015. View Article : Google Scholar : PubMed/NCBI
5	Xu HG, Yu YF, Zheng Q, Zhang W, Wang CD, Zhao XY, Tong WX, Wang H, Liu P and Zhang XL: Autophagy protects end plate chondrocytes from intermittent cyclic mechanical tension induced calcification. Bone. 66:232–239. 2014. View Article : Google Scholar : PubMed/NCBI
6	Pan J, Wang T, Wang L, Chen W and Song M: Cyclic strain-induced cytoskeletal rearrangement of human periodontal ligament cells via the Rho signaling pathway. PLoS One. 9:e915802014. View Article : Google Scholar : PubMed/NCBI
7	Xu HG, Ma MM, Zheng Q, Shen X, Wang H, Zhang SF, Xu JJ, Wang CD and Zhang XL: P120-catenin protects endplate chondrocytes from intermittent cyclic mechanical tension induced degeneration by inhibiting the expression of RhoA/ROCK-1 signaling pathway. Spine (Phila Pa 1976). 41:1261–1271. 2016. View Article : Google Scholar : PubMed/NCBI
8	Wang J, Wang CD, Zhang N, Tong WX, Zhang YF, Shan SZ, Zhang XL and Li QF: Mechanical stimulation orchestrates the osteogenic differentiation of human bone marrow stromal cells by regulating HDAC1. Cell Death Dis. 7:e22212016. View Article : Google Scholar : PubMed/NCBI
9	Zhang R, Mao P, Fu W, Pang XQ, Wang YY, Yang C, He WQ, Liu XQ and Li YM: Effects of cyclic stretch on the induction of the transdifferentiation in human lung epithelial cells. Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 25:455–459. 2013.(In Chinese). PubMed/NCBI
10	Witt F, Duda GN, Bergmann C and Petersen A: Cyclic mechanical loading enables solute transport and oxygen supply in bone healing: An in vitro investigation. Tissue Eng Part A. 20:486–493. 2014.PubMed/NCBI
11	Xu HG, Hu CJ, Wang H, Liu P, Yang XM, Zhang Y and Wang LT: Effects of mechanical strain on ANK, ENPP1 and TGF-β1 expression in rat endplate chondrocytes in vitro. Mol Med Rep. 4:831–835. 2011.PubMed/NCBI
12	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
13	Ferguson SJ and Steffen T: Biomechanics of the aging spine. Eur Spine J. 12 Suppl 2:S97–S103. 2003. View Article : Google Scholar : PubMed/NCBI
14	Alkhatib B, Rosenzweig DH, Krock E, Roughley PJ, Beckman L, Steffen T, Weber MH, Ouellet JA and Haglund L: Acute mechanical injury of the human intervertebral disc: Link to degeneration and pain. Eur Cells Mater. 28:98–111. 2014. View Article : Google Scholar
15	Sowa GA, Coelho JP, Vo NV, Pacek C, Westrick E and Kang JD: Cells from degenerative intervertebral discs demonstrate unfavorable responses to mechanical and inflammatory stimuli: A pilot study. Am J Phys Med Rehabil. 91:846–855. 2012. View Article : Google Scholar : PubMed/NCBI
16	Bruckner P and van der Rest M: Structure and function of cartilage collagens. Microsc Res Tech. 28:378–384. 1994. View Article : Google Scholar : PubMed/NCBI
17	Xu HG, Zheng Q, Song JX, Li J, Wang H, Liu P, Wang J, Wang CD and Zhang XL: Intermittent cyclic mechanical tension promotes endplate cartilage degeneration via canonical Wnt signaling pathway and E-cadherin/β-catenin complex cross-talk. Osteoarthritis Cartilage. 24:158–168. 2016. View Article : Google Scholar : PubMed/NCBI
18	Benya PD, Brown PD and Padilla SR: Microfilament modification by dihydrocytochalasin B causes retinoic acid-modulated chondrocytes to reexpress the differentiated collagen phenotype without a change in shape. J Cell Biol. 106:161–170. 1988. View Article : Google Scholar : PubMed/NCBI
19	Guilak F: Compression-induced changes in the shape and volume of the chondrocyte nucleus. J Biomech. 28:1529–1541. 1995. View Article : Google Scholar : PubMed/NCBI
20	Li S, Jia X, Duance VC and Blain EJ: The effects of cyclic tensile strain on the organisation and expression of cytoskeletal elements in bovine intervertebral disc cells: An in vitro study. Eur Cell Mater. 21:508–522. 2011. View Article : Google Scholar : PubMed/NCBI
21	Nofal GA and Knudson CB: Latrunculin and cytochalasin decrease chondrocyte matrix retention. J Histochem Cytochem. 50:1313–1324. 2002. View Article : Google Scholar : PubMed/NCBI