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European Journal of Medical Research
Published in: European Journal of Medical Research 1/2020

01-12-2020 | Research

Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures

Authors: Eliza S. Hartmann, Sabine Schluessel, Miriam I. Köhler, Felicitas Beck, Julia I. Redeker, Burkhard Summer, Veronika Schönitzer, Andreas Fottner, Susanne Mayer-Wagner

Published in: European Journal of Medical Research | Issue 1/2020

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Abstract

Introduction

Periprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplasties. Various studies have examined PPF behavior in monolayer culture systems. However, the periprosthetic tissue is a three-dimensional (3D) mesh, which allows the cells to interact in a multidirectional way. The expression of bone remodeling markers of fibroblast-like cells in a multilayer environment changes significantly versus monolayer cultures without the addition of particles or cytokine stimulation. Gene expression of bone remodeling markers was therefore compared in fibroblast-like cells from different origins and dermal fibroblasts under transwell culture conditions versus monolayer cultures.

Methods

PPFs from periprosthetic tissues (n = 12), osteoarthritic (OA) synovial fibroblast-like cells (SFs) (n = 6), and dermal fibroblasts (DFs) were cultured in monolayer (density 5.5 × 103/cm2) or multilayer cultures (density 8.5 × 105/cm2) for 10 or 21 days. Cultures were examined via histology, TRAP staining, immunohistochemistry (anti-S100a4), and quantitative real-time PCR.

Results

Fibroblast-like cells (PPFs/SFs) and dermal fibroblasts significantly increased the expression of RANKL and significantly decreased the expression of ALP, COL1A1, and OPG in multilayer cultures. PPFs and SFs in multilayer cultures further showed a higher expression of cathepsin K, MMP-13, and TNF-α. In multilayer PPF cultures, the mRNA level of TRAP was also found to be significantly increased.

Conclusion

The multilayer cultures are able to induce significant expression changes in fibroblast-like cells depending on the nature of cellular origin without the addition of any further stimulus. This system might be a useful tool to get more in vivo like results regarding fibroblast-like cell cultures.
Literature
1.
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
2.
Gallo J, Goodman SB, Konttinen YT, Raska M. Particle disease: biologic mechanisms of periprosthetic osteolysis in total hip arthroplasty. Innate Immun. 2013;19(2):213–24.CrossRefPubMed
3.
Gallo J, Goodman SB, Konttinen YT, Wimmer MA, Holinka M. Osteolysis around total knee arthroplasty: a review of pathogenetic mechanisms. Acta Biomater. 2013;9(9):8046–58.CrossRefPubMedPubMedCentral
4.
Koreny T, Tunyogi-Csapo M, Gal I, Vermes C, Jacobs JJ, Glant TT. The role of fibroblasts and fibroblast-derived factors in periprosthetic osteolysis. Arthr Rheum. 2006;54(10):3221–32.CrossRef
5.
Yao J, Glant TT, Lark MW, Mikecz K, Jacobs JJ, Hutchinson NI, et al. The potential role of fibroblasts in periprosthetic osteolysis: fibroblast response to titanium particles. J Bone Miner Res. 1995;10(9):1417–27.CrossRefPubMed
6.
Wei X, Zhang X, Flick LM, Drissi H, Schwarz EM, O’Keefe RJ. Titanium particles stimulate COX-2 expression in synovial fibroblasts through an oxidative stress-induced, calpain-dependent, NF-kappaB pathway. Am J Physiol Cell Physiol. 2009;297(2):C310–20.CrossRefPubMedPubMedCentral
7.
Pap T, Claus A, Ohtsu S, Hummel KM, Schwartz P, Drynda S, et al. Osteoclast-independent bone resorption by fibroblast-like cells. Arthr Res Ther. 2003;5(3):R163–73.CrossRef
8.
Mandelin J, Li TF, Hukkanen M, Liljestrom M, Salo J, Santavirta S, et al. Interface tissue fibroblasts from loose total hip replacement prosthesis produce receptor activator of nuclear factor-kappaB ligand, osteoprotegerin, and cathepsin K. J Rheumatol. 2005;32(4):713–20.PubMed
9.
Wagner S, Gollwitzer H, Wernicke D, Langer R, Siebenrock KA, Hofstetter W. Interface membrane fibroblasts around aseptically loosened endoprostheses express MMP-13. J Orthop Res. 2008;26(2):143–52.CrossRefPubMed
10.
Sabokbar A, Itonaga I, Sun SG, Kudo O, Athanasou NA. Arthroplasty membrane-derived fibroblasts directly induce osteoclast formation and osteolysis in aseptic loosening. J Orthop Res. 2005;23(3):511–9.CrossRefPubMed
11.
Qian YB, Zeng BF, Zhang XL, Jiang Y. Substance P stimulates production of interleukin 1 beta and tumor necrosis factor alpha in fibroblasts from hip periprosthetic membrane. J Arthroplasty. 2008;23(4):581–5.CrossRefPubMed
12.
Muller-Ladner U, Ospelt C, Gay S, Distler O, Pap T. Cells of the synovium in rheumatoid arthritis. Synovial fibroblasts. Arthr Res Ther. 2007;9(6):223.CrossRef
13.
Niedermeier M, Pap T, Korb A. Therapeutic opportunities in fibroblasts in inflammatory arthritis. Best Pract Res Clin Rheumatol. 2010;24(4):527–40.CrossRefPubMed
14.
15.
Sabater D, Fernandez-Lopez JA, Remesar X, Alemany M. The use of Transwells (TM) improves the rates of differentiation and growth of cultured 3T3L1 cells. Anal Bioanal Chem. 2013;405(16):5605–10.CrossRefPubMed
16.
Engler AJ, Sen S, Sweeney HL, Discher DE. Matrix elasticity directs stem cell lineage specification. Cell. 2006;126(4):677–89.CrossRefPubMed
17.
Koehler MI, Hartmann ES, Schluessel S, Beck F, Redeker JI, Schmitt B, et al. Impact of periprosthetic fibroblast-like cells on osteoclastogenesis in co-culture with peripheral blood mononuclear cells varies depending on culture system. Int J Mol Sci. 2019;20(10):2583.CrossRefPubMedCentral
18.
Wedemeyer C, Kauther MD, Hanenkamp S, Nuckel H, Bau M, Siffert W, et al. BCL2-938C > A and CALCA-1786T > C polymorphisms in aseptic loosened total hip arthroplasty. Eur J Med Res. 2009;14(6):250–5.CrossRefPubMedPubMedCentral
19.
Bloemen V, Schoenmaker T, de Vries TJ, Everts V. Direct cell-cell contact between periodontal ligament fibroblasts and osteoclast precursors synergistically increases the expression of genes related to osteoclastogenesis. J Cell Physiol. 2010;222(3):565–73.PubMed
20.
Granchi D, Amato I, Battistelli L, Ciapetti G, Pagani S, Avnet S, et al. Molecular basis of osteoclastogenesis induced by osteoblasts exposed to wear particles. Biomaterials. 2005;26(15):2371–9.CrossRefPubMed
21.
Hong H, Park YK, Choi MS, Ryu NH, Song DK, Suh SI, et al. Differential down-regulation of COX-2 and MMP-13 in human skin fibroblasts by glucosamine-hydrochloride. J Dermatol Sci. 2009;56(1):43–50.CrossRefPubMed
22.
Cordonnier T, Layrolle P, Gaillard J, Langonne A, Sensebe L, Rosset P, et al. 3D environment on human mesenchymal stem cells differentiation for bone tissue engineering. J Mater Sci. 2010;21(3):981–7.
23.
Vandesompele J, DePreter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002;3(7):RESEARCH0034.CrossRefPubMedPubMedCentral
24.
Waldele S, Koers-Wunrau C, Beckmann D, Korb-Pap A, Wehmeyer C, Pap T, et al. Deficiency of fibroblast activation protein alpha ameliorates cartilage destruction in inflammatory destructive arthritis. Arthr Res Ther. 2015;17:12.CrossRef
25.
Hartmann ES, Kohler MI, Huber F, Redeker JI, Schmitt B, Schmitt-Sody M, et al. Factors regulating bone remodelling processes in aseptic implant loosening. J Orthop Res. 2016;35(2):248–57.CrossRefPubMed
26.
Tsutsumi R, Xie C, Wei X, Zhang M, Zhang X, Flick LM, et al. PGE2 signaling through the EP4 receptor on fibroblasts upregulates RANKL and stimulates osteolysis. J Bone Miner Res. 2009;24(10):1753–62.CrossRefPubMedPubMedCentral
27.
Tomankova T, Kriegova E, Fillerova R, Luzna P, Ehrmann J, Gallo J. Comparison of periprosthetic tissues in knee and hip joints: differential expression of CCL3 and DC-STAMP in total knee and hip arthroplasty and similar cytokine profiles in primary knee and hip osteoarthritis. Osteoarthr Cartil. 2014;22(11):1851–60.CrossRef
28.
Koulouvaris P, Ly K, Ivashkiv LB, Bostrom MP, Nestor BJ, Sculco TP, et al. Expression profiling reveals alternative macrophage activation and impaired osteogenesis in periprosthetic osteolysis. J Orthop Res. 2008;26(1):106–16.CrossRefPubMed
29.
Ma GF, Ali A, Verzijl N, Hanemaaijer R, TeKoppele J, Konttinen YT, et al. Increased collagen degradation around loosened total hip replacement implants. Arthr Rheum. 2006;54(9):2928–33.CrossRef
30.
Everts V, Delaisse JM, Korper W, Niehof A, Vaes G, Beertsen W. Degradation of collagen in the bone-resorbing compartment underlying the osteoclast involves both cysteine-proteinases and matrix metalloproteinases. J Cell Physiol. 1992;150(2):221–31.CrossRefPubMed
31.
Everts V, Delaisse JM, Korper W, Beertsen W. Cysteine proteinases and matrix metalloproteinases play distinct roles in the subosteoclastic resorption zone. J Bone Miner Res. 1998;13(9):1420–30.CrossRefPubMed
32.
Hou WS, Li Z, Gordon RE, Chan K, Klein MJ, Levy R, et al. Cathepsin k is a critical protease in synovial fibroblast-mediated collagen degradation. Am J Pathol. 2001;159(6):2167–77.CrossRefPubMedPubMedCentral
33.
Hou WS, Li W, Keyszer G, Weber E, Levy R, Klein MJ, et al. Comparison of cathepsins K and S expression within the rheumatoid and osteoarthritic synovium. Arthr Rheum. 2002;46(3):663–74.CrossRef
34.
Takei I, Takagi M, Santavirta S, Ida H, Ishii M, Ogino T, et al. Messenger ribonucleic acid expression of 16 matrix metalloproteinases in bone-implant interface tissues of loose artificial hip joints. J Biomed Mater Res. 2000;52(4):613–20.CrossRefPubMed
35.
Delaisse JM, Andersen TL, Engsig MT, Henriksen K, Troen T, Blavier L. Matrix metalloproteinases (MMP) and cathepsin K contribute differently to osteoclastic activities. Microsc Res Tech. 2003;61(6):504–13.CrossRefPubMed
36.
Ninomiya JT, Struve JA, Stelloh CT, Toth JM, Crosby KE. Effects of hydroxyapatite particulate debris on the production of cytokines and proteases in human fibroblasts. J Orthop Res. 2001;19(4):621–8.CrossRefPubMed
37.
Costa-Rodrigues J, Teixeira CA, Sampaio P, Fernandes MH. Characterisation of the osteoclastogenic potential of human osteoblastic and fibroblastic conditioned media. J Cell Biochem. 2010;109(1):205–16.PubMed
38.
Weiss MJ, Ray K, Fallon MD, Whyte MP, Fedde KN, Lafferty MA, et al. Analysis of liver/bone/kidney alkaline phosphatase mRNA, DNA, and enzymatic activity in cultured skin fibroblasts from 14 unrelated patients with severe hypophosphatasia. Am J Hum Genet. 1989;44(5):686–94.PubMedPubMedCentral
39.
40.
Murdoch AD, Grady LM, Ablett MP, Katopodi T, Meadows RS, Hardingham TE. Chondrogenic differentiation of human bone marrow stem cells in transwell cultures: generation of scaffold-free cartilage. Stem cells. 2007;25(11):2786–96.CrossRefPubMed
41.
Madathil BK, Lin Q, Hew CL, Mohanty M. Hypoxia-like effect of cobalt chromium alloy micro particles on fibroblasts in vitro. J Orthop Res. 2010;28(10):1360–7.CrossRefPubMed
Metadata
Title
Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures
Authors
Eliza S. Hartmann
Sabine Schluessel
Miriam I. Köhler
Felicitas Beck
Julia I. Redeker
Burkhard Summer
Veronika Schönitzer
Andreas Fottner
Susanne Mayer-Wagner
Publication date
01-12-2020
Publisher
BioMed Central
Published in
European Journal of Medical Research / Issue 1/2020
Electronic ISSN: 2047-783X
DOI
https://doi.org/10.1186/s40001-020-00453-y

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