Effects of cyclic tension stress on the apoptosis of osteoclasts in vitro

Li,Fengbo; Sun,Xiaolei; Zhao,Bin; Ma,Jianxiong; Zhang,Yang; Li,Shuang; Li,Yanjun; Ma,Xinlong

doi:10.3892/etm.2015.2338

Effects of cyclic tension stress on the apoptosis of osteoclasts in vitro

Authors:
- Fengbo Li
- Xiaolei Sun
- Bin Zhao
- Jianxiong Ma
- Yang Zhang
- Shuang Li
- Yanjun Li
- Xinlong Ma
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Affiliations: Institute of Orthopedics, Tianjin Hospital, Tianjin 300211, P.R. China
Published online on: March 9, 2015 https://doi.org/10.3892/etm.2015.2338
Pages: 1955-1961

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Abstract

The aim of the present study was to investigate the effect of cyclic tension stress on osteoclast apoptosis in vitro using murine RAW264.7 cells treated with receptor activator of nuclear factor‑κB. Using the EF3200 mechanical testing instrument with BioDynamic bioreactor system, cultured osteoclasts which were seeded in a silicone rubber membrane load carrier, were loaded with periodic cyclic stretch microstrain. The induced osteoclasts were subjected to 0, 5, 10 and 15% stretch microstrain for 1 h daily for three days. The number of tartrate‑resistant acid phosphatase‑positive osteoclasts and the resorption area were assessed. Osteoclast apoptosis was detected by the Annexin V‑fluorescein isothiocyanate (FITC)/propidium iodide binding assay. The mRNA expression of Bcl‑2, Bax, caspase‑3 and cytochrome c was detected following force loading using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. Compared with the cells under no cyclic tension stress, the number of osteoclasts and the resorption area were increased in the cells under 10 and 15% stretch microstrain. The Annexin V binding assay showed that the early apoptosis rate of the 5, 10 and 15% stretch microstrain groups was decreased compared with that of the control group. RT‑qPCR results showed that the Bcl‑2/Bax ratio was significantly increased in the cells subjected to 5, 10 and 15% stretch microstrain compared with that in the control cells (P<0.05), while the expression of cytochrome c in the 10 and 15% stretch microstrain groups was decreased significantly (P<0.05). No significant difference was observed between the cytochrome c expression of the 5% stretch microstrain group and that of the control group (P>0.05). The expression of caspase‑3 in the 5, 10 and 15% stretch microstrain groups was decreased significantly compared with that in the control group (P<0.05). These data suggest that cyclic tension stress can inhibit apoptosis in osteoclasts, possibly by increasing the Bcl‑2/Bax ratio, inhibiting the activity of caspase‑3 and downregulating the expression of cytochrome c.

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1	Nakahama K: Cellular communications in bone homeostasis and repair. Cell Mol Life Sci. 67:4001–4009. 2010. View Article : Google Scholar : PubMed/NCBI
2	Liu H and Li B: p53 control of bone remodeling. J Cell Biochem. 111:529–534. 2010. View Article : Google Scholar : PubMed/NCBI
3	Ehrlich PJ and Lanyon LE: Mechanical strain and bone cell function: A review. Osteoporos Int. 13:688–700. 2002. View Article : Google Scholar : PubMed/NCBI
4	Chow JW, Jagger CJ and Chambers TJ: Reduction in dynamic indices of cancellous bone formation in rat tail vertebrae after caudal neurectomy. Calcif Tissue Int. 59:117–120. 1996. View Article : Google Scholar : PubMed/NCBI
5	Huiskes R, Ruimerman R, van Lenthe GH and Janssen JD: Effects of mechanical forces on maintenance and adaptation of form in trabecular bone. Nature. 405:704–706. 2000. View Article : Google Scholar : PubMed/NCBI
6	Rubin CT and Lanyon LE: Kappa Delta Award paper. Osteoregulatory nature of mechanical stimuli: Function as a determinant for adaptive remodeling in bone. J Orthop Res. 5:300–310. 1987. View Article : Google Scholar : PubMed/NCBI
7	Jee WS, Li XJ and Schaffler MB: Adaptation of diaphyseal structure with aging and increased mechanical usage in the adult rat: A histomorphometrical and biomechanical study. Anat Rec. 230:332–338. 1991. View Article : Google Scholar : PubMed/NCBI
8	Turner CH, Akhter MP, Raab DM, Kimmel DB and Recker RR: A noninvasive, in vivo model for studying strain adaptive bone modeling. Bone. 12:73–79. 1991. View Article : Google Scholar : PubMed/NCBI
9	Chambers TJ, Evans M, Gardner TN, Turner-Smith A and Chow JW: Induction of bone formation in rat tail vertebrae by mechanical loading. Bone Miner. 20:167–178. 1993. View Article : Google Scholar : PubMed/NCBI
10	Forwood MR and Turner CH: Skeletal adaptations to mechanical usage: Results from tibial loading studies in rats. Bone. 17(4 Suppl): 197S–205S. 1995. View Article : Google Scholar : PubMed/NCBI
11	Mosley JR, March BM, Lynch J and Lanyon LE: Strain magnitude related changes in whole bone architecture in growing rats. Bone. 20:191–198. 1997. View Article : Google Scholar : PubMed/NCBI
12	Cowin SC, Moss-Salentijn L and Moss ML: Candidates for the mechanosensory system in bone. J Biomech Eng. 113:191–197. 1991. View Article : Google Scholar : PubMed/NCBI
13	Weinbaum S, Cowin SC and Zeng Y: A model for the excitation of osteocytes by mechanical loading-induced bone fluid shear stresses. J Biomech. 27:339–360. 1994. View Article : Google Scholar : PubMed/NCBI
14	Kaspar D, Seidl W, Neidlinger-Wilke C, et al: Proliferation of human-derived osteoblast-like cells depends on the cycle number and frequency of uniaxial strain. J Biomech. 35:873–880. 2002. View Article : Google Scholar : PubMed/NCBI
15	Kaspar D, Seidl W, Neidlinger-Wilke C, et al: Dynamic cell stretching increases human osteoblast proliferation and CICP synthesis but decreases osteocalcin synthesis and alkaline phosphatase activity. J Biomech. 33:45–51. 2000. View Article : Google Scholar : PubMed/NCBI
16	Neidlinger-Wilke C, Wilke HJ and Claes L: Cyclic stretching of human osteoblasts affects proliferation and metabolism: A new experimental method and its application. J Orthop Res. 12:70–78. 1994. View Article : Google Scholar : PubMed/NCBI
17	Chen YJ, Huang CH, Lee IC, et al: Effects of cyclic mechanical stretching on the mRNA expression of tendon/ligament-related and osteoblast-specific genes in human mesenchymal stem cells. Connect Tissue Res. 49:7–14. 2008. View Article : Google Scholar : PubMed/NCBI
18	Hara F, Fukuda K, Ueno M, et al: Pertussis toxin-sensitive G proteins as mediators of stretch-induced decrease in nitric-oxide release of osteoblast-like cells. J Orthop Res. 17:593–597. 1999. View Article : Google Scholar : PubMed/NCBI
19	Searby ND, Steele CR and Globus RK: Influence of increased mechanical loading by hypergravity on the microtubule cytoskeleton and prostaglandin E2 release in primary osteoblasts. Am J Physiol Cell Physiol. 289:C148–C158. 2005. View Article : Google Scholar : PubMed/NCBI
20	Kanno T, Takahashi T, Tsujisawa T, et al: Mechanical stress-mediated Runx2 activation is dependent on Ras/ERK1/2 MAPK signaling in osteoblasts. J Cell Biochem. 101:1266–1277. 2007. View Article : Google Scholar : PubMed/NCBI
21	Boyle WJ, Simonet WS and Lacey DL: Osteoclast differentiation and activation. Nature. 423:337–342. 2003. View Article : Google Scholar : PubMed/NCBI
22	Quinn JM and Gillespie MT: Modulation of osteoclast formation. Biochem Biophys Res Commun. 328:739–745. 2005. View Article : Google Scholar : PubMed/NCBI
23	Suzuki N, Yoshimura Y, Deyama Y, et al: Mechanical stress directly suppresses osteoclast differentiation in RAW264. 7 cells. Int J Mol Med. 21:291–296. 2008.
24	Kurata K, Uemura T, Nemoto A, et al: Mechanical strain effect on bone-resorbing activity and messenger RNA expressions of marker enzymes in isolated osteoclast culture. J Bone Miner Res. 16:722–730. 2001. View Article : Google Scholar : PubMed/NCBI
25	Weinstein RS and Manolagas SC: Apoptosis and osteoporosis. Am J Med. 108:153–164. 2000. View Article : Google Scholar : PubMed/NCBI
26	Abe K, Yoshimura Y, Deyama Y, et al: Effects of bisphosphonates on osteoclastogenesis in RAW264. 7 cells. Int J Mol Med. 29:1007–1015. 2012.
27	Mdel M Arriero, Ramis JM, Perelló J and Monjo M: Inositol hexakisphosphate inhibits osteoclastogenesis on RAW 264.7 cells and human primary osteoclasts. PLoS One. 7:e431872012. View Article : Google Scholar : PubMed/NCBI
28	Osdoby P, Martini MC and Caplan AI: Isolated osteoclasts and their presumed progenitor cells, the monocyte, in culture. J Exp Zool. 224:331–344. 1982. View Article : Google Scholar : PubMed/NCBI
29	Zallone A Zambonin, Teti A and Primavera MV: Isolated osteoclasts in primary culture: First observations on structure and survival in culture media. Anat Embryol (Berl). 165:405–413. 1982. View Article : Google Scholar : PubMed/NCBI
30	Tezuka K, Tezuka Y, Maejima A, et al: Molecular cloning of a possible cysteine proteinase predominantly expressed in osteoclasts. J Biol Chem. 269:1106–1109. 1994.PubMed/NCBI
31	Udagawa N, Takahashi N, Akatsu T, et al: The bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoclast-like cell differentiation in cocultures with mouse spleen cells. Endocrinology. 125:1805–1813. 1989. View Article : Google Scholar : PubMed/NCBI
32	Collin-Osdoby P, Oursler MJ, Webber D and Osdoby P: Osteoclast-specific monoclonal antibodies coupled to magnetic beads provide a rapid and efficient method of purifying avian osteoclasts. J Bone Miner Res. 6:1353–1365. 1991. View Article : Google Scholar : PubMed/NCBI
33	Quinn JM, Neale S, Fujikawa Y, et al: Human osteoclast formation from blood monocytes, peritoneal macrophages, and bone marrow cells. Calcif Tissue Int. 62:527–531. 1998. View Article : Google Scholar : PubMed/NCBI
34	Duncan RL and Turner CH: Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tissue Int. 57:344–358. 1995. View Article : Google Scholar : PubMed/NCBI
35	Skerry TM and Suva LJ: Investigation of the regulation of bone mass by mechanical loading: From quantitative cytochemistry to gene array. Cell Biochem Funct. 21:223–229. 2003. View Article : Google Scholar : PubMed/NCBI
36	Kroemer G, Galluzzi L and Brenner C: Mitochondrial membrane permeabilization in cell death. Physiol Rev. 87:99–163. 2007. View Article : Google Scholar : PubMed/NCBI
37	Adams JM and Cory S: Bcl-2-regulated apoptosis: Mechanism and therapeutic potential. Curr Opin Immunol. 19:488–496. 2007. View Article : Google Scholar : PubMed/NCBI
38	Yin XM: Bid, a BH3-only multi-functional molecule, is at the cross road of life and death. Gene. 369:7–19. 2006. View Article : Google Scholar : PubMed/NCBI
39	Certo M, Del Gaizo Moore V, Nishino M, et al: Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. Cancer Cell. 9:351–365. 2006. View Article : Google Scholar : PubMed/NCBI
40	Kuwana T and Newmeyer DD: Bcl-2-family proteins and the role of mitochondria in apoptosis. Curr Opin Cell Biol. 15:691–699. 2003. View Article : Google Scholar : PubMed/NCBI
41	Hellebrand EE and Varbiro G: Development of mitochondrial permeability transition inhibitory agents: A novel drug target. Drug Discov Ther. 4:54–61. 2010.PubMed/NCBI
42	Jilka RL, Noble B and Weinstein RS: Osteocyte apoptosis. Bone. 54:264–271. 2013. View Article : Google Scholar : PubMed/NCBI
43	Timmer JC and Salvesen GS: Caspase substrates. Cell Death Differ. 14:66–72. 2007. View Article : Google Scholar : PubMed/NCBI
44	Dai C, Zhang B, Liu X, et al: Pyrimethamine sensitizes pituitary adenomas cells to temozolomide through cathepsin B-dependent and caspase-dependent apoptotic pathways. Int J Cancer. 133:1982–1993. 2013. View Article : Google Scholar : PubMed/NCBI
45	Cryns V and Yuan J: Proteases to die for. Genes Dev. 12:1551–1570. 1998. View Article : Google Scholar : PubMed/NCBI