Lentivirus-mediated TNF-α gene silencing and overexpression of osteoprotegerin inhibit titanium particle-induced inflammatory response and osteoclastogenesis in vitro

Peng,Li; Wang,Hongzhi; Song,Keguan; Wang,Hai; Liu,Ping

doi:10.3892/mmr.2015.4620

January-2016 Volume 13 Issue 1

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January-2016 Volume 13 Issue 1

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Article

Lentivirus-mediated TNF-α gene silencing and overexpression of osteoprotegerin inhibit titanium particle-induced inflammatory response and osteoclastogenesis in vitro

Authors:
- Li Peng
- Hongzhi Wang
- Keguan Song
- Hai Wang
- Ping Liu
View Affiliations / Copyright

Affiliations: The Third Department of Orthopaedics, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Orthopaedics, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350004, P.R. China, Department of Immunology, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China
Pages: 1010-1018
|
Published online on: November 27, 2015

https://doi.org/10.3892/mmr.2015.4620
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Abstract

Macrophages and osteoclasts release proinflammatory factors and promote osteoclastogenesis following the phagocytosis of wear particles. During this pathological process, receptor of nuclear factor κB ligand (RANKL) and tumor necrosis factor (TNF)-α are critical factors contributing to resorption and the inflammatory response. The present study aimed to construct recombination lentivirus vectors carrying TNF-α small interfering (si)RNA and osteoprotegerin (OPG) cDNA, and to examine the effects of Lenti‑siTNFα‑OPG on the wear particle‑induced inflammatory response and osteoclastogenesis in a titanium (Ti) particle‑induced‑inflammatory response cell model. Lenti‑siTNFα‑OPG vectors were constructed and transnfected into RAW264.7 and MC3T3‑E1 cells, respectively, prior to particle stimulation. The protein levels of TNF‑α, OPG and RANKL were evaluated using western blot analysis and enzyme‑linked immunosorbent assays, and the mRNA expression levels of the inflammatory factors, TNF‑α, interleukin (IL)‑1β and IL‑6, as well as OPG and RANKL, were measured using reverse transcription‑quantitative polymerase chain reaction analysis. The activity of alkaline phosphatase (ALP) was examined using an ALP kit. In the presence of the Lenti‑siTNFα‑OPG vector, the mRNA expression levels of the inflammatory factors and RANKL were downregulated, as were the protein levels of TNF‑α. The mRNA expression and protein levels of OPG were upregulated, and ALP activity was increased. These findings suggested that Lenti‑siTNFα‑OPG transfection inhibited the wear particle‑induced inflammatory response and osteoclastogenesis, which warrants further investigation for the prevention and/or treatment of wear particle-induced osteolysis.

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1	Ollivere B, Wimhurst JA, Clark IM and Donell ST: Current concepts in osteolysis. J Bone Joint Surg Br. 94:10–15. 2012. View Article : Google Scholar : PubMed/NCBI
2	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
3	Harris WH: Wear and periprosthetic osteolysis: The problem. Clin Orthop Relat Res. 66–70. 2001. View Article : Google Scholar
4	Schmalzried TP, Jasty M and Harris WH: Periprosthetic bone loss in total hip arthroplasty. Polyethylene wear debris and the concept of the effective joint space. J Bone Joint Surg Am. 74:849–863. 1992.PubMed/NCBI
5	Agarwal S: Osteolysis-basic science, incidence and diagnosis. Curr Orthopaed. 18:220–231. 2004. View Article : Google Scholar
6	Purdue PE, Koulouvaris P, Nestor BJ and Sculco TP: The central role of wear debris in periprosthetic osteolysis. HSS J. 2:102–113. 2006. View Article : Google Scholar
7	Abu-Amer Y, Darwech I and Clohisy JC: Aseptic loosening of total joint replacements: Mechanisms underlying osteolysis and potential therapies. Arthritis Res Ther. 9(Suppl 1): S62007. View Article : Google Scholar : PubMed/NCBI
8	Boyce BF and Xing L: Biology of RANK, RANKL and osteoprotegerin. Arthritis Res Ther. 9(Suppl 1): S12007. View Article : Google Scholar :
9	Takahashi N, Maeda K, Ishihara A, Uehara S and Kobayashi Y: Regulatory mechanism of osteoclastogenesis by RANKL and Wnt signals. Front Biosci (Landmark Ed). 16:21–30. 2011. View Article : Google Scholar
10	Kwan Tat S, Padrines M, Théoleyre S, Heymann D and Fortun Y: IL-6, RANKL, TNF-alpha/IL-1: Interrelations in bone resorption pathophysiology. Cytokine Growth Factor Rev. 15:49–60. 2004. View Article : Google Scholar : PubMed/NCBI
11	Akisue T, Bauer TW, Farver CF and Mochida Y: The effect of particle wear debris on NFkappaB activation and pro-inflammatory cytokine release in differentiated THP-1 cells. J Biomed Mater Res. 59:507–515. 2002. View Article : Google Scholar : PubMed/NCBI
12	Greenfield EM and Bechtold J; Implant Wear Symposium 2007 Biologic Work Group: What other biologic and mechanical factors might contribute to osteolysis? J Am Acad Orthop Sur. 16(Suppl): S56–S62. 2008.
13	Abbas S, Zhang YH, Clohisy JC and Abu-Amer Y: Tumor necrosis factor-alpha inhibits pre-osteoblast differentiation through its type-1 receptor. Cytokine. 22:33–41. 2003. View Article : Google Scholar : PubMed/NCBI
14	Xu J, Wu HF, Ang ES, Yip K, Woloszyn M, Zheng MH and Tan RX: NF-kappaB modulators in osteolytic bone diseases. Cytokine Growth Factor Rev. 20:7–17. 2009. View Article : Google Scholar
15	Yoshino T, Yamaguchi M, Shimizu M, Yamada K and Kasai K: TNF-α aggravates the progression of orthodontically-induced inflammatory root resorption in the presence of RANKL. J Hard Tissue Biol. 23:155–162. 2014. View Article : Google Scholar
16	Froelich S, Tai A and Wang P: Lentiviral vectors for immune cells targeting. Immunopharmacol Immunotoxicol. 32:208–218. 2010. View Article : Google Scholar : PubMed/NCBI
17	Rucci N, Rufo A, Alamanou M and Teti A: Modeled microgravity stimulates osteoclastogenesis and bone resorption by increasing osteoblast RANKL/OPG ratio. J Cell Biochem. 100:464–473. 2007. View Article : Google Scholar
18	Philpott A, Weston-Simons JS, Grammatopoulos G, Bejon P, Gill HS, McLardy-Smith P, Gundle R, Murray DW and Pandit H: Predictive outcomes of revision total hip replacement-A consecutive series of 1176 patients with a minimum 10-year follow-up. Maturitas. 77:185–190. 2014. View Article : Google Scholar
19	Goodman SB, Trindade M, Ma T, Genovese M and Smith RL: Pharmacologic modulation of periprosthetic osteolysis. Clin Orthop Relat Res. 39–45. 2005. View Article : Google Scholar : PubMed/NCBI
20	Wang CT, Lin YT, Chiang BL, Lee SS and Hou SM: Over-expression of receptor activator of nuclear factor-kappaB ligand (RANKL), inflammatory cytokines and chemokines in periprosthetic oste-olysis of loosened total hip arthroplasty. Biomaterials. 31:77–82. 2010. View Article : Google Scholar
21	Teitelbaum SL: Osteoclasts; culprits in inflammatory osteolysis. Arthritis Res Ther. 8(201)2006.
22	Ritchlin CT, Haas-Smith SA, Li P, Hicks DG and Schwarz EM: Mechanisms of TNF-alpha and RANKL mediated osteoclasto-genesis and bone resorption in psoriatic arthritis. J Clin Invest. 111:821–831. 2003. View Article : Google Scholar : PubMed/NCBI
23	He X, Andersson G, Lindgren U and Li Y: Resveratrol prevents RANKL-induced osteoclast differentiation of murine osteoclast progenitor RAW264.7 cells through inhibition of ROS production. Biochem Biophys Res Commun. 401:356–362. 2010. View Article : Google Scholar : PubMed/NCBI
24	Adapala NS, Barbe MF, Langdon WY, Nakamura MC, Tsygankov AY and Sanjay A: The loss of Cbl-phosphatidylinositol 3-kinase interaction perturbs RANKL-mediated signaling, inhibiting bone resorption and promoting osteoclast survival. J Biol Chem. 285:36745–36758. 2010. View Article : Google Scholar : PubMed/NCBI
25	Inose H, Ochi H, Kimura A, Fujita K, Xu R, Sato S, Iwasaki M, Sunamura S, Takeuchi Y, Fukumoto S, et al: A microRNA regulatory mechanism of osteoblast differentiation. Proc Natl Acad Sci USA. 106:20794–20799. 2009. View Article : Google Scholar : PubMed/NCBI
26	Bonnelye E, Chabadel A, Saltel F and Jurdic P: Dual effect of strontium ranelate: Stimulation of osteoblast differentiation and inhibition of osteoclast formation and resorption in vitro. Bone. 42:129–138. 2008. View Article : Google Scholar
27	Almeida M, Han L, Ambrogini E, Weinstein RS and Manolagas SC: Glucocorticoids and tumor necrosis factor α increase oxidative stress and suppress Wnt protein signaling in osteoblasts. J Biol Chem. 286:44326–44335. 2011. View Article : Google Scholar : PubMed/NCBI
28	Pivonka P, Zimak J, Smith DW, Gardiner BS, Dunstan CR, Sims NA, Martin TJ and Mundy GR: Theoretical investigation of the role of the RANK-RANKL-OPG system in bone remodeling. J Theor Biol. 262:306–316. 2010. View Article : Google Scholar
29	Dong L, Wang R, Zhu YA, Wang C, Diao H, Zhang C, Zhao J and Zhang J: Antisense oligonucleotide targeting TNF-alpha can suppress Co-Cr-Mo particle-induced osteolysis. J Orthop Res. 26:1114–1120. 2008. View Article : Google Scholar : PubMed/NCBI
30	Goater JJ, O'Keefe RJ, Rosier RN, Puzas JE and Schwarz EM: Efficacy of ex vivo OPG gene therapy in preventing wear debris induced osteolysis. J Orthop Res. 20:169–173. 2002. View Article : Google Scholar : PubMed/NCBI
31	Kay MA, Glorioso JC and Naldini L: Viral vectors for gene therapy: The art of turning infectious agents into vehicles of therapeutics. Nat Med. 7:33–40. 2001. View Article : Google Scholar : PubMed/NCBI