Excessive production of mitochondrion‑derived reactive oxygen species induced by titanium ions leads to autophagic cell death of osteoblasts via the SIRT3/SOD2 pathway

Wang,Siqian; Yang,Jingyuan; Lin,Tingting; Huang,Shengbing; Ma,Jianfeng; Xu,Xin

doi:10.3892/mmr.2020.11094

Excessive production of mitochondrion‑derived reactive oxygen species induced by titanium ions leads to autophagic cell death of osteoblasts via the SIRT3/SOD2 pathway

Authors:
- Siqian Wang
- Jingyuan Yang
- Tingting Lin
- Shengbing Huang
- Jianfeng Ma
- Xin Xu
View Affiliations

Affiliations: Department of Implantology, School and Hospital of Stomatology, Shandong University, Shandong Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong 250012, P.R. China, Department of Prothodontics, School and Hospital of Stomatology, Wenzhou Medical University, Wenzhou, Zhejiang 325027, P.R. China
Published online on: April 24, 2020 https://doi.org/10.3892/mmr.2020.11094
Pages: 257-264
Copyright: © Wang 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 incidence of peri-implant bone loss is high, and is a difficult condition to treat. Previous studies have shown that titanium (Ti) ions released from implants can lead to osteoblast cell damage, but the specific mechanisms have not been elucidated. The present study established a Ti ion damage osteoblast cell model. The levels of mitochondrion‑derived reactive oxygen species (mROS) and autophagy, cell viability and the sirtuin 3 (SIRT3)/superoxide dismutase 2 (SOD2) pathway were examined in this model. It was found that Ti ions decreased osteoblast viability. Moreover, with increased Ti ion concentration, the expression levels of microtubule associated protein 1 light chain 3α (LC3) progressively increased, P62 decreased, autophagic flow increased and mROS levels increased. After the addition of an autophagy inhibitor Bafilomycin A1 and Mito‑TEMPO, a mitochondrial antioxidant, the production of mROS was inhibited, the level of autophagy was decreased and cell activity was improved. In addition, with increased Ti ion concentration, the activity of SOD2 decreased, the acetylation level of SOD2 increased, the SIRT3 mRNA and protein expression levels decreased, and the activity of SIRT3 was significantly decreased. Furthermore, it was demonstrated that SIRT3 overexpression reduced the acetylation of SOD2 and increased the activity of SOD2, as well as reducing the production of mROS and the expression level of LC3, thus increasing cell viability. Therefore, the present results suggested that excessive production of mROS induced by Ti ions led to autophagic cell death of osteoblasts, which is dependent on the SIRT3/SOD2 pathway.

View Figures

View References

1	Anselme K: Osteoblast adhesion on biomaterials. Biomaterials. 21:667–681. 2000. View Article : Google Scholar : PubMed/NCBI
2	Anselme K: Biomaterials and interface with bone. Osteoporos Int. 22:2037–2042. 2011. View Article : Google Scholar : PubMed/NCBI
3	Davies JE: Mechanisms of endosseous integration. Int J Prosthodont. 11:391–401. 1998.PubMed/NCBI
4	Davies JE: Understanding peri-implant endosseous healing. J Dent Educ. 67:932–949. 2003.PubMed/NCBI
5	Hermann JS, Buser D, Schenk RK, Schoolfield JD and Cochran DL: Biologic Width around one- and two-piece titanium implants. Clin Oral Implants Res. 12:559–571. 2001. View Article : Google Scholar : PubMed/NCBI
6	Lindhe J and Meyle J; Group D of European Workshop on Periodontology, : Peri-implant diseases: Consensus Report of the Sixth European Workshop on Periodontology. J Clin Periodontol. 35 (Suppl):282–285. 2008. View Article : Google Scholar : PubMed/NCBI
7	Brånemark PI, Adell R, Albrektsson T, Lekholm U, Lundkvist S and Rockler B: Osseointegrated titanium fixtures in the treatment of edentulousness. Biomaterials. 4:25–28. 1983. View Article : Google Scholar : PubMed/NCBI
8	Wachi T, Shuto T, Shinohara Y, Matono Y and Makihira S: Release of titanium ions from an implant surface and their effect on cytokine production related to alveolar bone resorption. Toxicology. 327:1–9. 2015. View Article : Google Scholar : PubMed/NCBI
9	Schwarz F, Sahm N, Mihatovic I, Golubovic V and Becker J: Surgical therapy of advanced ligature-induced peri-implantitis defects: Cone-beam computed tomographic and histological analysis. J Clin Periodontol. 38:939–949. 2011. View Article : Google Scholar : PubMed/NCBI
10	Kumazawa R, Watari F, Takashi N, Tanimura Y, Uo M and Totsuka Y: Effects of Ti ions and particles on neutrophil function and morphology. Biomaterials. 23:3757–3764. 2002. View Article : Google Scholar : PubMed/NCBI
11	Wennerberg A, Ide-Ektessabi A, Hatkamata S, Sawase T, Johansson C, Albrektsson T, Martinelli A, Södervall U and Odelius H: Titanium release from implants prepared with different surface roughness. Clin Oral Implants Res. 15:505–512. 2004. View Article : Google Scholar : PubMed/NCBI
12	Ellingsen JE, Johansson CB, Wennerberg A and Holmén A: Improved retention and bone-tolmplant contact with fluoride-modified titanium implants. Int J Oral Maxillofac Implants. 19:659–666. 2004.PubMed/NCBI
13	Pajarinen J, Kouri VP, Jämsen E, Li TF, Mandelin J and Konttinen YT: The response of macrophages to titanium particles is determined by macrophage polarization. Acta Biomater. 9:9229–9240. 2013. View Article : Google Scholar : PubMed/NCBI
14	Obando-Pereda GA, Fischer L and Stach-Machado DR: Titanium and zirconia particle-induced pro-inflammatory gene expression in cultured macrophages and osteolysis, inflammatory hyperalgesia and edema in vivo. Life Sci. 97:96–106. 2014. View Article : Google Scholar : PubMed/NCBI
15	Brettin BT, Grosland NM, Qian F, Southard KA, Stuntz TD, Morgan TA, Marshall SD and Southard TE: Bicortical vs monocortical orthodontic skeletal anchorage. Am J Orthod Dentofacial Orthop. 134:625–635. 2008. View Article : Google Scholar : PubMed/NCBI
16	Bitar D and Parvizi J: Biological response to prosthetic debris. World J Orthop. 6:172–189. 2015. View Article : Google Scholar : PubMed/NCBI
17	Grunow M and Schöpp W: [Determination of the activity of superoxide dismutase in terms of rea substrate conversion]. Biomed Biochim Acta. 48:185–199. 1989.(In German). PubMed/NCBI
18	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
19	Klionsky DJ, Elazar Z, Seglen PO and Rubinsztein DC: Does bafilomycin A1 block the fusion of autophagosomes with lysosomes? Autophagy. 4:849–850. 2008. View Article : Google Scholar : PubMed/NCBI
20	Sena LA and Chandel NS: Physiological roles of mitochondrial reactive oxygen species. Mol Cell. 48:158–167. 2012. View Article : Google Scholar : PubMed/NCBI
21	Lambert AJ and Brand MD: Reactive oxygen species production by mitochondria. Methods Mol Biol. 554:165–181. 2009. View Article : Google Scholar : PubMed/NCBI
22	Tao R, Vassilopoulos A, Parisiadou L, Yan Y and Gius D: Regulation of MnSOD enzymatic activity by Sirt3 connects the mitochondrial acetylome signaling networks to aging and carcinogenesis. Antioxid Redox Signal. 20:1646–1654. 2014. View Article : Google Scholar : PubMed/NCBI
23	Chen Y, Zhang J, Lin Y, Lei Q, Guan KL, Zhao S and Xiong Y: Tumour suppressor SIRT3 deacetylates and activates manganese superoxide dismutase to scavenge ROS. EMBO Rep. 12:534–541. 2011. View Article : Google Scholar : PubMed/NCBI
24	Tao R, Coleman MC, Pennington JD, Ozden O, Park SH, Jiang H, Kim HS, Flynn CR, Hill S, Hayes McDonald W, et al: Sirt3-mediated deacetylation of evolutionarily conserved lysine 122 regulates MnSOD activity in response to stress. Mol Cell. 40:893–904. 2010. View Article : Google Scholar : PubMed/NCBI
25	Di Tucci AA, Murru R, Alberti D, Rabault B, Deplano S and Angelucci E: Correction of anemia in a transfusion-dependent patient with primary myelofibrosis receiving iron chelation therapy with deferasirox (Exjade, ICL670). Eur J Haematol. 78:540–542. 2007. View Article : Google Scholar : PubMed/NCBI
26	Messa E, Cilloni D, Messa F, Arruga F, Roetto A and Saglio G: Deferasirox treatment improved the hemoglobin level and decreased transfusion requirements in four patients with the myelodysplastic syndrome and primary myelofibrosis. Acta Haematol. 120:70–74. 2008. View Article : Google Scholar : PubMed/NCBI
27	Angelucci E, Santini V, Di Tucci AA, Quaresmini G, Finelli C, Volpe A, Quarta G, Rivellini F, Sanpaolo G, Cilloni D, et al: Deferasirox for transfusion-dependent patients with myelodysplastic syndromes: Safety, efficacy, and beyond (GIMEMA MDS0306 Trial). Eur J Haematol. 92:527–536. 2014. View Article : Google Scholar : PubMed/NCBI
28	Jacobs JJ, Gilbert JL and Urban RM: Corrosion of metal orthopaedic implants. J Bone Joint Surg Am. 80:268–282. 1998. View Article : Google Scholar : PubMed/NCBI
29	Gallo J, Kamínek P, Tichá V, Riháková P and Ditmar R: Particle disease. A comprehensive theory of periprosthetic osteolysis: A review. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 146:21–28. 2002. View Article : Google Scholar : PubMed/NCBI
30	Nakagawa M, Matsuya S and Udoh K: Effects of fluoride and dissolved oxygen concentrations on the corrosion behavior of pure titanium and titanium alloys. Dent Mater J. 21:83–92. 2002. View Article : Google Scholar : PubMed/NCBI
31	Matono Y, Nakagawa M, Matsuya S, Ishikawa K and Terada Y: Corrosion behavior of pure titanium and titanium alloys in various concentrations of Acidulated Phosphate Fluoride (APF) solutions. Dent Mater J. 25:104–112. 2006. View Article : Google Scholar : PubMed/NCBI
32	Roehlecke C, Witt M, Kasper M, Schulze E, Wolf C, Hofer A and Funk RW: Synergistic effect of titanium alloy and collagen type I on cell adhesion, proliferation and differentiation of osteoblast-like cells. Cells Tissues Organs. 168:178–187. 2001. View Article : Google Scholar : PubMed/NCBI
33	Terheyden H, Lang NP, Bierbaum S and Stadlinger B: Osseointegration--communication of cells. Clin Oral Implants Res. 23:1127–1135. 2012. View Article : Google Scholar : PubMed/NCBI
34	Rajendrakumar SK, Revuri V, Samidurai M, Mohapatra A, Lee JH, Ganesan P, Jo J, Lee YK and Park IK: Peroxidase-mimicking nanoassembly mitigates lipopolysaccharide-induced endotoxemia and cognitive damage in the brain by impeding inflammatory signaling in macrophages. Nano Lett. 18:6417–6426. 2018. View Article : Google Scholar : PubMed/NCBI
35	Landgraeber S, Jäger M, Jacobs JJ and Hallab NJ: The pathology of orthopedic implant failure is mediated by innate immune system cytokines. Mediators Inflamm. 2014:1851502014. View Article : Google Scholar : PubMed/NCBI
36	Li X, Ma XY, Feng YF, Ma ZS, Wang J, Ma TC, Qi W, Lei W and Wang L: Osseointegration of chitosan coated porous titanium alloy implant by reactive oxygen species-mediated activation of the PI3K/AKT pathway under diabetic conditions. Biomaterials. 36:44–54. 2015. View Article : Google Scholar : PubMed/NCBI
37	Feng YF, Wang L, Zhang Y, Li X, Ma ZS, Zou JW, Lei W and Zhang ZY: Effect of reactive oxygen species overproduction on osteogenesis of porous titanium implant in the present of diabetes mellitus. Biomaterials. 34:2234–2243. 2013. View Article : Google Scholar : PubMed/NCBI
38	Wang L, Zhao X, Wei BY, Liu Y, Ma XY, Wang J, Cao PC, Zhang Y, Yan YB, Lei W, et al: Insulin improves osteogenesis of titanium implants under diabetic conditions by inhibiting reactive oxygen species overproduction via the PI3K-Akt pathway. Biochimie. 108:85–93. 2015. View Article : Google Scholar : PubMed/NCBI
39	Klionsky DJ, Abdelmohsen K, Abe A, Abedin MDJ, Abeliovich H, Arozena AA, Adachi H, Adams CM, Adams PD, Adeli K, et al: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 12:1–222. 2016. View Article : Google Scholar : PubMed/NCBI
40	Liu Y and Levine B: Autosis and autophagic cell death: The dark side of autophagy. Cell Death Differ. 22:367–376. 2015. View Article : Google Scholar : PubMed/NCBI
41	Essick EE and Sam F: Oxidative stress and autophagy in cardiac disease, neurological disorders, aging and cancer. Oxid Med Cell Longev. 3:168–177. 2010. View Article : Google Scholar : PubMed/NCBI
42	Zhang T, Li Y, Park KA, Byun HS, Won M, Jeon J, Lee Y, Seok JH, Choi SW, Lee SH, et al: Cucurbitacin induces autophagy through mitochondrial ROS production which counteracts to limit caspase-dependent apoptosis. Autophagy. 8:559–576. 2012. View Article : Google Scholar : PubMed/NCBI
43	Shen W, Tian C, Chen H, Yang Y, Zhu D, Gao P and Liu J: Oxidative stress mediates chemerin-induced autophagy in endothelial cells. Free Radic Biol Med. 55:73–82. 2013. View Article : Google Scholar : PubMed/NCBI
44	Dai DF and Rabinovitch P: Mitochondrial oxidative stress mediates induction of autophagy and hypertrophy in angiotensin-II treated mouse hearts. Autophagy. 7:917–918. 2011. View Article : Google Scholar : PubMed/NCBI
45	Kim Y, Kim YS, Kim DE, Lee JS, Song JH, Kim HG, Cho DH, Jeong SY, Jin DH, Jang SJ, et al: BIX-01294 induces autophagy-associated cell death via EHMT2/G9a dysfunction and intracellular reactive oxygen species production. Autophagy. 9:2126–2139. 2013. View Article : Google Scholar : PubMed/NCBI
46	Liang Q, Benavides GA, Vassilopoulos A, Gius D, Darley-Usmar V and Zhang J: Bioenergetic and autophagic control by Sirt3 in response to nutrient deprivation in mouse embryonic fibroblasts. Biochem J. 454:249–257. 2013. View Article : Google Scholar : PubMed/NCBI
47	Pi H, Xu S, Reiter RJ, Guo P, Zhang L, Li Y, Li M, Cao Z, Tian L, Xie J, et al: SIRT3-SOD2-mROS-dependent autophagy in cadmium-induced hepatotoxicity and salvage by melatonin. Autophagy. 11:1037–1051. 2015. View Article : Google Scholar : PubMed/NCBI
48	Lee J, Giordano S and Zhang J: Autophagy, mitochondria and oxidative stress: Cross-talk and redox signalling. Biochem J. 441:523–540. 2012. View Article : Google Scholar : PubMed/NCBI
49	Zorov DB, Juhaszova M and Sollott SJ: Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol Rev. 94:909–950. 2014. View Article : Google Scholar : PubMed/NCBI
50	Zhu Y, Park SH, Ozden O, Kim HS, Jiang H, Vassilopoulos A, Spitz DR and Gius D: Exploring the electrostatic repulsion model in the role of Sirt3 in directing MnSOD acetylation status and enzymatic activity. Free Radic Biol Med. 53:828–833. 2012. View Article : Google Scholar : PubMed/NCBI
51	Zeng L, Yang Y, Hu Y, Sun Y, Du Z, Xie Z, Zhou T and Kong W: Age-related decrease in the mitochondrial sirtuin deacetylase Sirt3 expression associated with ROS accumulation in the auditory cortex of the mimetic aging rat model. PLoS One. 9:e880192014. View Article : Google Scholar : PubMed/NCBI
52	Li M, Chiu JF, Mossman BT and Fukagawa NK: Down-regulation of manganese-superoxide dismutase through phosphorylation of FOXO3a by Akt in explanted vascular smooth muscle cells from old rats. J Biol Chem. 281:40429–40439. 2006. View Article : Google Scholar : PubMed/NCBI
53	Niska K, Pyszka K, Tukaj C, Wozniak M, Radomski MW and Inkielewicz-Stepniak I: Titanium dioxide nanoparticles enhance production of superoxide anion and alter the antioxidant system in human osteoblast cells. Int J Nanomedicine. 10:1095–1107. 2015.PubMed/NCBI
54	Li Y, Wang W, Wu Q, Li Y, Tang M, Ye B and Wang D: Molecular control of TiO₂-NPs toxicity formation at predicted environmental relevant concentrations by Mn-SODs proteins. PLoS One. 7:e446882012. View Article : Google Scholar : PubMed/NCBI
55	Bánréti A, Sass M and Graba Y: The emerging role of acetylation in the regulation of autophagy. Autophagy. 9:819–829. 2013. View Article : Google Scholar : PubMed/NCBI
56	Papanicolaou KN, O'Rourke B and Foster DB: Metabolism leaves its mark on the powerhouse: Recent progress in post-translational modifications of lysine in mitochondria. Front Physiol. 5:3012014. View Article : Google Scholar : PubMed/NCBI
57	Hebert AS, Dittenhafer-Reed KE, Yu W, Bailey DJ, Selen ES, Boersma MD, Carson JJ, Tonelli M, Balloon AJ, Higbee AJ, et al: Calorie restriction and SIRT3 trigger global reprogramming of the mitochondrial protein acetylome. Mol Cell. 49:186–199. 2013. View Article : Google Scholar : PubMed/NCBI
58	Giralt A and Villarroya F: SIRT3, a pivotal actor in mitochondrial functions: Metabolism, cell death and aging. Biochem J. 444:1–10. 2012. View Article : Google Scholar : PubMed/NCBI
59	Kim H, Lee YD, Kim HJ, Lee ZH and Kim HH: SOD2 and Sirt3 Control Osteoclastogenesis by Regulating Mitochondrial ROS. J Bone Miner Res. 32:397–406. 2017. View Article : Google Scholar : PubMed/NCBI