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BMC Musculoskeletal Disorders
Published in: BMC Musculoskeletal Disorders 1/2016

Open Access 01-12-2016 | Research article

Lentivirus-mediated short hairpin RNA interference targeting TNF-alpha in macrophages inhibits particle-induced inflammation and osteolysis in vitro and in vivo

Authors: Chu-qiang Qin, Dong-sheng Huang, Chi Zhang, Bin Song, Jian-bin Huang, Yue Ding

Published in: BMC Musculoskeletal Disorders | Issue 1/2016

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Abstract

Background

Aseptic loosening is a significant impediment to joint implant longevity. Prosthetic wear particles are postulated to play a central role in the onset and progression of periprosthetic osteolysis, leading to aseptic loosening of the prosthesis.

Methods

We investigated the inhibitory effects of a lentivirus-mediated short hairpin RNA that targets the TNF-alpha gene on the particle-induced inflammatory and osteolytic changes via macrophages both in vitro and in vivo. An siRNA sequence targeting the mouse TNF-alpha gene from four candidates, transcribed in vitro, was screened and identified. A lentivirus vector expressing short hairpin RNA (shRNA) was then constructed in order to facilitate efficient expression of TNF-alpha-siRNA. Lentivirus-mediated shRNA was transduced into cells of the mouse macrophage line RAW 264.7. Ceramic and titanium particles were introduced 24 h after lentivirus transduction to stimulate cells. TNF-alpha expression, represented by both mRNA and protein levels, was quantified with real-time PCR and ELISA at all time intervals. Lentivirus-mediated shRNA suspension was locally administered into the murine calvarial model, followed by local injection of particles. A multi-slice spiral CT scan was used to evaluate the osteolysis of the calvaria by detecting the width of the cranial sutures.

Results

Macrophages developed pseudopods when co-cultured with particles. Lentivirus-mediated shRNA was shown to effectively inhibit the expression of TNF-alpha at both the mRNA and protein levels in RAW 264.7. The multi-slice spiral CT scan showed that the lentivirus-mediated shRNA significantly suppressed osteolysis of mouse calvaria.

Conclusions

Our investigation highlighted the results that lentivirus-mediated shRNA targeting the TNF-alpha gene successfully inhibited particle-induced inflammatory and osteolytic changes both in vitro and in vivo. Therefore, lentivirus-mediated gene therapy may provide a novel therapeutic approach to aseptic joint loosening.
Literature
1.
Keener JD, Callaghan JJ, Goetz DD, Pederson DR, Sullivan PM, Johnston RC. Twenty-five-year results after Charnley total hip arthroplasty in patients less than fifty years old: a concise follow-up of a previous report. J Bone Joint Surg Am. 2003;85-A(6):1066–72.CrossRefPubMed
2.
Warme BA, Epstein NJ, Trindade MC, Miyanishi K, Ma T, Saket RR, et al. Proinflammatory mediator expression in a novel murine model of titanium-particle-induced intramedullary inflammation. J Biomed Mater Res B Appl Biomater. 2004;71(2):360–6.CrossRefPubMed
3.
Buly RL, Huo MH, Salvati E, Brien W, Bansal M. Titanium wear debris in failed cemented total hip arthroplasty. An analysis of 71 cases. J Arthroplasty. 1992;7(3):315–23.CrossRefPubMed
4.
Goodman SB. Allograft alternatives: bone substitutes and beyond. Orthopedics. 2010;33(9):661.PubMed
5.
Yang F, Wu W, Cao L, Huang Y, Zhu Z, Tang T, et al. Pathways of macrophage apoptosis within the interface membrane in aseptic loosening of prostheses. Biomaterials. 2011;32(35):9159–67.CrossRefPubMed
6.
Bauer TW, Shanbhag AS. Are there biological markers of wear? J Am Acad Orthop Surg. 2008;16 Suppl 1:S68–71.CrossRefPubMed
7.
Ren PG, Irani A, Huang Z, Ma T, Biswal S, Goodman SB. Continuous infusion of UHMWPE particles induces increased bone macrophages and osteolysis. Clin Orthop Relat Res. 2011;469(1):113–22.CrossRefPubMed
8.
Jin S, Park JY, Hong JM, Kim TH, Shin HI, Park EK, et al. Inhibitory effect of (−)-epigallocatechin gallate on titanium particle-induced TNF-alpha release and in vivo osteolysis. Exp Mol Med. 2011;43(7):411–8.CrossRefPubMedPubMedCentral
9.
Childs LM, Goater JJ, O’Keefe RJ, Schwarz EM. Efficacy of etanercept for wear debris-induced osteolysis. J Bone Miner Res. 2001;16(2):338–47.CrossRefPubMed
10.
Miettinen SS, Jaatinen J, Pelttari A, Lappalainen R, Monkkonen J, Venesmaa PK, et al. Effect of locally administered zoledronic acid on injury-induced intramembranous bone regeneration and osseointegration of a titanium implant in rats. J Orthop Sci. 2009;14(4):431–6.CrossRefPubMed
11.
Friedl G, Radl R, Stihsen C, Rehak P, Aigner R, Windhager R. The effect of a single infusion of zoledronic acid on early implant migration in total hip arthroplasty. A randomized, double-blind, controlled trial. J Bone Joint Surg Am. 2009;91(2):274–81.CrossRefPubMed
12.
Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 1998;391(6669):806–11.CrossRefPubMed
13.
Manjunath NKPL. Interfering antiviral immunity: application, subversion, hope? Trends Immunol. 2006;27(7):328–35.CrossRefPubMed
14.
Morris KRJ. Lentiviral-mediated delivery of siRNAs for antiviral therapy. Gene Ther. 2006.
15.
Pluta K, Kacprzak MM. Use of HIV as a gene transfer vector. Acta Biochim Pol. 2009;56(4):531–95.PubMed
16.
Sinn PAAB. Lentivirus vector can be readministered to nasal epithelia without blocking immune responses. J Virol. 2008.
17.
Escors D, Breckpot K. Lentiviral vectors in gene therapy: their current status and future potential. Arch Immunol Ther Exp (Warsz). 2010;58(2):107–19.CrossRef
18.
Wedemeyer C, Xu J, Neuerburg C, Landgraeber S, Malyar NM, von Knoch F, et al. Particle-induced osteolysis in three-dimensional micro-computed tomography. Calcif Tissue Int. 2007;81(5):394–402.CrossRefPubMed
19.
Beck RT, Illingworth KD, Saleh KJ. Review of periprosthetic osteolysis in total joint arthroplasty: an emphasis on host factors and future directions. J Orthop Res. 2012;30(4):541–6.CrossRefPubMed
20.
Catelas I, Jacobs JJ. Biologic activity of wear particles. Instr Course Lect. 2010;59:3–16.PubMed
21.
Shanbhag AS. Use of bisphosphonates to improve the durability of total joint replacements. J Am Acad Orthop Surg. 2006;14(4):215–25.CrossRefPubMed
22.
Nich C, Takakubo Y, Pajarinen J, Ainola M, Salem A, Sillat T, et al. Macrophages-Key cells in the response to wear debris from joint replacements. J Biomed Mater Res a. 2013;101(10):3033–45.CrossRefPubMedPubMedCentral
23.
Mabilleau G, Pandit H, Jinnah RH, Sabokbar A. Biological response to common surface bearings used in orthopaedics. J Surg Orthop Adv. 2008;17(1):34–9.PubMed
24.
Ding Y, Qin CQ, Fu YR, Xu J, Huang DS. In vitro comparison of the biological activity of alumina ceramic and titanium particles associated with aseptic loosening. Biomed Mater. 2012;7(4):45019.CrossRef
25.
26.
Wang H, Jia TH, Zacharias N, Gong W, Du HX, Wooley PH et al. Combination gene therapy targeting on interleukin-1beta and RANKL for wear debris-induced aseptic loosening. Gene Ther. 2012;20:128–35.CrossRefPubMedPubMedCentral
27.
Li P, Schwarz EM, O’Keefe RJ, Ma L, Looney RJ, Ritchlin CT, et al. Systemic tumor necrosis factor alpha mediates an increase in peripheral CD11bhigh osteoclast precursors in tumor necrosis factor alpha-transgenic mice. Arthritis Rheum. 2004;50(1):265–76.CrossRefPubMed
28.
Boyce BF, Li P, Yao Z, Zhang Q, Badell IR, Schwarz EM, et al. TNF-alpha and pathologic bone resorption. Keio J Med. 2005;54(3):127–31.CrossRefPubMed
29.
Popivanova BK, Kitamura K, Wu Y, Kondo T, Kagaya T, Kaneko S, et al. Blocking TNF-alpha in mice reduces colorectal carcinogenesis associated with chronic colitis. J Clin Invest. 2008;118(2):560–70.PubMedPubMedCentral
30.
Deng Y, Wang CC, Choy KW, Du Q, Chen J, Wang Q, et al. Therapeutic potentials of gene silencing by RNA interference: principles, challenges, and new strategies. Gene. 2014;538(2):217–27.CrossRefPubMed
31.
Lundstrom K. Latest development in viral vectors for gene therapy. Trends Biotechnol. 2003;21(3):117–22.CrossRefPubMed
32.
Primo MN, Bak RO, Mikkelsen JG. Lentiviral vectors for cutaneous RNA managing. Exp Dermatol. 2012;21(3):162–70.CrossRefPubMed
33.
34.
Utzschneider S, Becker F, Grupp TM, Sievers B, Paulus A, Gottschalk O, et al. Inflammatory response against different carbon fiber-reinforced PEEK wear particles compared with UHMWPE in vivo. Acta Biomater. 2010;6(11):4296–304.CrossRefPubMed
35.
Merkel KD, Erdmann JM, McHugh KP, Abu-Amer Y, Ross FP, Teitelbaum SL. Tumor necrosis factor-alpha mediates orthopedic implant osteolysis. Am J Pathol. 1999;154(1):203–10.CrossRefPubMedPubMedCentral
36.
Mao X, Pan X, Cheng T, Zhang X. Therapeutic potential of the proteasome inhibitor Bortezomib on titanium particle-induced inflammation in a murine model. Inflammation. 2012;35(3):905–12.CrossRefPubMed
37.
Yang SY, Yu H, Gong W, Wu B, Mayton L, Costello R, et al. Murine model of prosthesis failure for the long-term study of aseptic loosening. J Orthop Res. 2007;25(5):603–11.CrossRefPubMed
38.
Khatri N, Rathi M, Baradia D, Trehan S, Misra A. In vivo delivery aspects of miRNA, shRNA and siRNA. Crit Rev Ther Drug Carrier Syst. 2012;29(6):487–527.CrossRefPubMed
39.
Vicentini FT, Borgheti-Cardoso LN, Depieri LV, de Macedo MD, Abelha TF, Petrilli R, et al. Delivery systems and local administration routes for therapeutic siRNA. Pharm Res. 2013;30(4):915–31.CrossRefPubMed
Metadata
Title
Lentivirus-mediated short hairpin RNA interference targeting TNF-alpha in macrophages inhibits particle-induced inflammation and osteolysis in vitro and in vivo
Authors
Chu-qiang Qin
Dong-sheng Huang
Chi Zhang
Bin Song
Jian-bin Huang
Yue Ding
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Musculoskeletal Disorders / Issue 1/2016
Electronic ISSN: 1471-2474
DOI
https://doi.org/10.1186/s12891-016-1290-6

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