Cyclic mechanical tension reinforces DNA damage and activates the p53-p21-Rb pathway to induce premature senescence of nucleus pulposus cells

Feng,Chencheng; Yang,Minghui; Zhang,Yang; Lan,Minghong; Huang,Bo; Liu,Huan; Zhou,Yue

doi:10.3892/ijmm.2018.3522

Cyclic mechanical tension reinforces DNA damage and activates the p53-p21-Rb pathway to induce premature senescence of nucleus pulposus cells

Authors:
- Chencheng Feng
- Minghui Yang
- Yang Zhang
- Minghong Lan
- Bo Huang
- Huan Liu
- Yue Zhou
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Affiliations: Department of Orthopedics, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, P.R. China
Published online on: February 28, 2018 https://doi.org/10.3892/ijmm.2018.3522
Pages: 3316-3326
Copyright: © Feng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Intervertebral disc (IVD) degeneration (IDD) is a widely recognized contributor to low back pain. Mechanical stress is a crucial etiological factor of IDD. During the process of IDD, a vicious circle is formed between abnormal mechanical stress and the damage of disc structure and function. Notably, the pathological process of IDD is mediated by the phenotypic shift of IVD cells from an extracellular matrix anabolic phenotype to a catabolic and pro-inflammatory phenotype. Therefore, the effects of mechanical stress on the initiation and progression of IDD depend on the mechanobiology of IVD cells. Recently, disc cell senescence was identified as a new hallmark of IDD. However, the senescent response of disc cells to mechanical stress remains unknown. In this study, we found that prolonged exposure of cyclic mechanical tension (CMT) with unphysiological magnitude generated by the Flexercell tension system markedly induced premature senescence of nucleus pulposus (NP) cells. CMT augmented the DNA damage of NP cells, but did not affect the redox homeostasis of NP cells. Moreover, the p53-p21-retinoblastoma protein (Rb) pathway was activated by CMT to mediate the CMT-induced premature senescence of NP cells. The findings are beneficial to understanding the mechanism of disc cell senescence and the mechanobiology of disc cells further. It suggests that prolonged abnormal mechanical stress accelerates the establishment and progression of disc cell senescence and consequently impairs the structural and functional homeostasis of IVDs to cause IDD. Preventing the pro-senescent effect of mechanical stress on IVD cells is a promising approach to delay the process of IDD.

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1	Vos T, Flaxman AD, Naghavi M, Lozano R, Michaud C, Ezzati M, Shibuya K, Salomon JA, Abdalla S, Aboyans V, et al: Years lived with disability (YLDs) for 1160 sequelae of 289 diseases and injuries 1990-2010: A systematic analysis for the Global Burden of Disease Study 2010. Lancet. 380:2163–2196. 2012. View Article : Google Scholar : PubMed/NCBI
2	Hong J, Reed C, Novick D and Happich M: Costs associated with treatment of chronic low back pain: An analysis of the UK General Practice Research Database. Spine. 38:75–82. 2013. View Article : Google Scholar
3	Cheung KM, Karppinen J, Chan D, Ho DW, Song YQ, Sham P, Cheah KS, Leong JC and Luk KD: Prevalence and pattern of lumbar magnetic resonance imaging changes in a population study of one thousand forty-three individuals. Spine. 34:934–940. 2009. View Article : Google Scholar : PubMed/NCBI
4	Takatalo J, Karppinen J, Niinimäki J, Taimela S, Näyhä S, Mutanen P, Sequeiros RB, Kyllönen E and Tervonen O: Does lumbar disc degeneration on magnetic resonance imaging associate with low back symptom severity in young Finnish adults? Spine. 36:2180–2189. 2011. View Article : Google Scholar : PubMed/NCBI
5	Roberts S, Evans H, Trivedi J and Menage J: Histology and pathology of the human intervertebral disc. J Bone Joint Surg Am. 88(Suppl 2): 10–14. 2006.PubMed/NCBI
6	Kanayama M, Togawa D, Takahashi C, Terai T and Hashimoto T: Cross-sectional magnetic resonance imaging study of lumbar disc degeneration in 200 healthy individuals. J Neurosurg Spine. 11:501–507. 2009. View Article : Google Scholar : PubMed/NCBI
7	Battié MC, Videman T, Kaprio J, Gibbons LE, Gill K, Manninen H, Saarela J and Peltonen L: The Twin Spine Study: Contributions to a changing view of disc degeneration. Spine J. 9:47–59. 2009. View Article : Google Scholar
8	Wang D, Nasto LA, Roughley P, Leme AS, Houghton AM, Usas A, Sowa G, Lee J, Niedernhofer L, Shapiro S, et al: Spine degeneration in a murine model of chronic human tobacco smokers. Osteoarthritis and cartilage/OARS. Osteoarthritis Res Soc. 20:896–905. 2012. View Article : Google Scholar
9	Stirling A, Worthington T, Rafiq M, Lambert PA and Elliott TS: Association between sciatica and Propionibacterium acnes. Lancet. 357:2024–2025. 2001. View Article : Google Scholar : PubMed/NCBI
10	Park EY and Park JB: Dose- and time-dependent effect of high glucose concentration on viability of notochordal cells and expression of matrix degrading and fibrotic enzymes. Int Orthop. 37:1179–1186. 2013. View Article : Google Scholar : PubMed/NCBI
11	Neidlinger-Wilke C1, Galbusera F, Pratsinis H, Mavrogonatou E, Mietsch A, Kletsas D and Wilke HJ: Mechanical loading of the intervertebral disc: from the macroscopic to the cellular level. Eur Spine J. 23(Suppl 3): S333–S343. 2014. View Article : Google Scholar
12	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 and cartilage/OARS. Osteoarthritis Res Soc. 23:1057–1070. 2015. View Article : Google Scholar
13	Setton LA and Chen J: Mechanobiology of the intervertebral disc and relevance to disc degeneration. J Bone Joint Surg Am. 88(Suppl 2): 52–57. 2006.PubMed/NCBI
14	Risbud MV and Shapiro IM: Role of cytokines in intervertebral disc degeneration: Pain and disc content. Nat Rev Rheumatol. 10:44–56. 2014. View Article : Google Scholar
15	Vo NV, Hartman RA, Yurube T, Jacobs LJ, Sowa GA and Kang JD: Expression and regulation of metalloproteinases and their inhibitors in intervertebral disc aging and degeneration. Spine J. 13:331–341. 2013. View Article : Google Scholar : PubMed/NCBI
16	Miyamoto H, Doita M, Nishida K, Yamamoto T, Sumi M and Kurosaka M: Effects of cyclic mechanical stress on the production of inflammatory agents by nucleus pulposus and anulus fibrosus derived cells in vitro. Spine. 31:4–9. 2006. View Article : Google Scholar : PubMed/NCBI
17	Gilbert HT, Hoyland JA and Millward-Sadler SJ: The response of human anulus fibrosus cells to cyclic tensile strain is frequency-dependent and altered with disc degeneration. Arthritis Rheum. 62:3385–3394. 2010. View Article : Google Scholar : PubMed/NCBI
18	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
19	Zhang YH, Zhao CQ, Jiang LS and Dai LY: Cyclic stretch-induced apoptosis in rat annulus fibrosus cells is mediated in part by endoplasmic reticulum stress through nitric oxide production. Eur Spine J. 20:1233–1243. 2011. View Article : Google Scholar : PubMed/NCBI
20	Roberts S, Evans EH, Kletsas D, Jaffray DC and Eisenstein SM: Senescence in human intervertebral discs. Eur Spine J. 15(Suppl 3): S312–S316. 2006. View Article : Google Scholar : PubMed/NCBI
21	Le Maitre CL, Freemont AJ and Hoyland JA: Accelerated cellular senescence in degenerate intervertebral discs: A possible role in the pathogenesis of intervertebral disc degeneration. Arthritis Res Ther. 9:R452007. View Article : Google Scholar : PubMed/NCBI
22	Wang F, Cai F, Shi R, Wang XH and Wu XT: Aging and age related stresses: A senescence mechanism of intervertebral disc degeneration. Osteoarthritis and cartilage/OARS. Osteoarthritis Res Soc. 24:398–408. 2016. View Article : Google Scholar
23	Feng C, Liu H, Yang M, Zhang Y, Huang B and Zhou Y: Disc cell senescence in intervertebral disc degeneration: Causes and molecular pathways. Cell Cycle. 15:1674–1684. 2016. View Article : Google Scholar : PubMed/NCBI
24	Dimozi A, Mavrogonatou E, Sklirou A and Kletsas D: Oxidative stress inhibits the proliferation, induces premature senescence and promotes a catabolic phenotype in human nucleus pulposus intervertebral disc cells. Eur Cell Mater. 30:89–103. 2015. View Article : Google Scholar : PubMed/NCBI
25	Jeong SW, Lee JS and Kim KW: In vitro lifespan and senescence mechanisms of human nucleus pulposus chondrocytes. Spine J. 14:499–504. 2014. View Article : Google Scholar
26	Purmessur D, Walter BA, Roughley PJ, Laudier DM, Hecht AC and Iatridis J: A role for TNFα in intervertebral disc degeneration: A non-recoverable catabolic shift. Biochem Biophys Res Commun. 433:151–156. 2013. View Article : Google Scholar : PubMed/NCBI
27	Xing QJ, Liang QQ, Bian Q, Ding DF, Cui XJ, Shi Q and Wang YJ: Leg amputation accelerates senescence of rat lumbar intervertebral discs. Spine. 35:E1253–E1261. 2010. View Article : Google Scholar : PubMed/NCBI
28	Liang QQ, Zhou Q, Zhang M, Hou W, Cui XJ, Li CG, Li TF, Shi Q and Wang YJ: Prolonged upright posture induces degenerative changes in intervertebral discs in rat lumbar spine. Spine. 33:2052–2058. 2008. View Article : Google Scholar : PubMed/NCBI
29	O'Connell GD, Johannessen W, Vresilovic EJ and Elliott DM: Human internal disc strains in axial compression measured noninvasively using magnetic resonance imaging. Spine. 32:2860–2868. 2007. View Article : Google Scholar
30	Nerurkar NL, Elliott DM and Mauck RL: Mechanical design criteria for intervertebral disc tissue engineering. J Biomech. 43:1017–1030. 2010. View Article : Google Scholar : PubMed/NCBI
31	Broberg KB: On the mechanical behaviour of intervertebral discs. Spine. 8:151–165. 1983. 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
33	Hiyama A, Sakai D, Risbud MV, Tanaka M, Arai F, Abe K and Mochida J: Enhancement of intervertebral disc cell senescence by WNT/β-catenin signaling-induced matrix metalloproteinase expression. Arthritis Rheum. 62:3036–3047. 2010. View Article : Google Scholar : PubMed/NCBI
34	Keorochana G, Johnson JS, Taghavi CE, Liao JC, Lee KB, Yoo JH, Ngo SS and Wang JC: The effect of needle size inducing degeneration in the rat caudal disc: Evaluation using radiograph, magnetic resonance imaging, histology, and immunohistochemistry. Spine J. 10:1014–1023. 2010. View Article : Google Scholar : PubMed/NCBI
35	Jun JI and Lau LF: The matricellular protein CCN1 induces fibroblast senescence and restricts fibrosis in cutaneous wound healing. Nat Cell Biol. 12:676–685. 2010. View Article : Google Scholar : PubMed/NCBI
36	Gruber HE, Watts JA, Hoelscher GL, Bethea SF, Ingram JA, Zinchenko NS and Hanley EN Jr: Mitochondrial gene expression in the human annulus: In vivo data from annulus cells and selectively harvested senescent annulus cells. Spine J. 11:782–791. 2011. View Article : Google Scholar : PubMed/NCBI
37	Kim KW, Chung HN, Ha KY, Lee JS and Kim YY: Senescence mechanisms of nucleus pulposus chondrocytes in human inter-vertebral discs. Spine J. 9:658–666. 2009. View Article : Google Scholar : PubMed/NCBI
38	Gruber HE, Ingram JA, Norton HJ and Hanley EN Jr: Senescence in cells of the aging and degenerating intervertebral disc: Immunolocalization of senescence-associated beta-galactosidase in human and sand rat discs. Spine. 32:321–327. 2007. View Article : Google Scholar : PubMed/NCBI
39	Sakai D and Andersson GB: Stem cell therapy for intervertebral disc regeneration: Obstacles and solutions. Nat Rev Rheumatol. 11:243–256. 2015. View Article : Google Scholar : PubMed/NCBI
40	Binch AL, Cole AA, Breakwell LM, Michael AL, Chiverton N, Cross AK and Le Maitre CL: Expression and regulation of neurotrophic and angiogenic factors during human intervertebral disc degeneration. Arthritis Res Ther. 16:4162014. View Article : Google Scholar : PubMed/NCBI
41	Binch AL, Cole AA, Breakwell LM, Michael AL, Chiverton N, Creemers LB, Cross AK and Le Maitre CL: Nerves are more abundant than blood vessels in the degenerate human intervertebral disc. Arthritis Res Ther. 17:3702015. View Article : Google Scholar : PubMed/NCBI
42	Khan IM, Gilbert SJ, Caterson B, Sandell LJ and Archer CW: Oxidative stress induces expression of osteoarthritis markers procollagen IIA and 3B3(-) in adult bovine articular cartilage. Osteoarthritis and cartilage/OARS. Osteoarthritis Res Soc. 16:698–707. 2008. View Article : Google Scholar
43	Martin JA, Brown TD, Heiner AD and Buckwalter JA: Chondrocyte senescence, joint loading and osteoarthritis. Clin Orthop Relat Res. 427(Suppl): S96–S103. 2004. View Article : Google Scholar
44	Xu HG, Zheng Q, Song JX, Li J, Wang H, Liu P, Wang J, Wang C and Zhang X: Intermittent cyclic mechanical tension promotes endplate cartilage degeneration via canonical Wnt signaling pathway and E-cadherin/beta-catenin complex cross-talk. Osteoarthritis and cartilage/OARS. Osteoarthritis Res Soc. 24:158–168. 2016. View Article : Google Scholar
45	Xiao L, Xu HG, Wang H, Liu P, Liu C, Shen X, Zhang T and Xu YM: Intermittent cyclic mechanical tension promotes degeneration of endplate cartilage via the nuclear factor-κB signaling pathway: An in vivo study. Orthop Surg. 8:393–399. 2016. View Article : Google Scholar : PubMed/NCBI
46	Hunter CJ, Matyas JR and Duncan NA: Cytomorphology of notochordal and chondrocytic cells from the nucleus pulposus: A species comparison. J Anat. 205:357–362. 2004. View Article : Google Scholar : PubMed/NCBI
47	Rufai A, Benjamin M and Ralphs JR: The development of fibro-cartilage in the rat intervertebral disc. Anat Embryol (Berl). 192:53–62. 1995. View Article : Google Scholar
48	Stevens JW, Kurriger GL, Carter AS and Maynard JA: CD44 expression in the developing and growing rat intervertebral disc. Dev Dyn. 219:381–390. 2000. View Article : Google Scholar : PubMed/NCBI
49	Alini M, Eisenstein SM, Ito K, Little C, Kettler AA, Masuda K, Melrose J, Ralphs J, Stokes I and Wilke HJ: Are animal models useful for studying human disc disorders/degeneration? Eur Spine J. 17:2–19. 2008. View Article : Google Scholar
50	Daly C, Ghosh P, Jenkin G, Oehme D and Goldschlager T: A Review of animal models of intervertebral disc degeneration: Pathophysiology, regeneration, and translation to the clinic. BioMed Res Int. 2016:59521652016. View Article : Google Scholar : PubMed/NCBI