In vivo, cytokines noncovalently bind to the extracellular matrix (ECM), to facilitate intimate interactions with cellular receptors and potentiate biological activity. Development of a biomaterial that simulates this type of physiological binding and function is an exciting proposition for designing controlled advanced delivery systems for simulating in vivo conditions in vitro. We have decorated silk protein with sulfonated moieties through diazonium coupling reactions to noncovalently immobilize pro-inflammatory cytokines interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in such a biomimetic manner. After adsorption of the cytokines to the diazonium-modified silk matrix, constant release of cytokines up to at least 3 days was demonstrated, as an initial step to simulate an osteoarthritic (OA) microenvironment in vitro. Matrix-embedded cytokines induced the formation of multiple elongated processes in chondrocytes in vitro, akin to what is seen in OA cartilage in vivo. Gene expression profiles with this in vitro tissue model of OA showed significant similarities to profiles from explanted OA cartilage tissues collected from patients who underwent total knee replacement surgery. The common markers of OA, including COL, MMP, TIMP, ADAMTS, and metallothioneins, were upregulated at least 35-fold in the in vitro model when compared to the control—non-OA in vitro generated tissue-engineered cartilage. The microarray data were validated by reverse transcriptase–polymerase chain reaction. Mechanistically, protein interaction studies indicated that TNF-α and IL-1β synergistically controlled the equilibrium between MMPs and their inhibitors, TIMPs, resulting in ECM degradation through the MAPK pathway. This study offers a promising initial step toward establishing a relevant in vitro OA disease model, which can be further modified to assess signaling mechanisms, responses to cell or drug treatments and patient-specific features.
Get full access to this article
View all access options for this article.
References
1.
AndersonD.D., ChubinskayaS., GuilakF., MartinJ.A., OegemaT.R., OlsonS.A.et al.Post-traumatic osteoarthritis: Improved understanding and opportunities for early intervention. J Orthop Res, 29:802. 2011.
2.
LiY., XuL., OlsenB.R.Lessons from genetic forms of osteoarthritis for the pathogenesis of the disease. Osteoarthritis Cartilage, 15:1101. 2007.
3.
MartinJ.A., BuckwalterJ.A.Telomere erosion and senescence in human articular cartilage chondrocytes. J Gerontol A Biol Sci Med Sci, 56:B172. 2001.
4.
AignerT., RoseJ., MartinJ., BuckwalterJ.Aging theories of primary osteoarthritis: from epidemiology to molecular biology. Rejuvenation Res, 7:134. 2004.
5.
ValdesA.M., LoughlinJ., OeneM.V., ChapmanK., SurdulescuG.L., DohertyM.et al.Sex and ethnic differences in the association of ASPN, CALM1, COL2A1, COMP, and FRZB with genetic susceptibility to osteoarthritis of the knee. Arthritis Rheum, 56:137. 2007.
6.
TakahashiH., NakajimaM., OzakiK., TanakaT., KamataniN., IkegawaS.Prediction model for knee osteoarthritis based on genetic and clinical information. Arthritis Res Ther, 12:R187. 2010.
7.
PritzkerK. P. H.Osteoarthritis: joint instability and OA: do animal models provide insights?Nat Rev Rheumatol, 7:444. 2011.
8.
PelletierJ.P., Martel-PelletierJ, AbramsonS.B.Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis Rheum, 44:1237. 2001.
9.
SmithM.D., TriantafillouS., ParkerA., YoussefP.P., ColemanM.Synovial membrane inflammation and cytokine production in patients with early osteoarthritis. J Rheumatol, 24:365. 1997.
10.
BondesonJ., WainwrightS.D., LauderS., AmosN., HughesC.E.The role of synovial macrophages and macrophage-produced cytokines in driving aggrecanases, matrix metalloproteinases, and other destructive and inflammatory responses in osteoarthritis. Arthritis Res Ther, 8:R187. 2006.
11.
TowleC.A., HungH.H., BonassarL.J., TreadwellB.V., ManghamD.G.Detection of interleukin-1 in the cartilage of patients with osteoarthritis: a possible autocrine/paracrine role in pathogenesis. Osteoarthritis Cartilage, 5:293. 1997.
12.
OllivierreF., GublerU., TowleC.A., LaurencinC., TreadwellB.V.Expression of IL-1 genes in human and bovine chondrocytes: a mechanism for autocrine control of cartilage matrix degradation. Biochem Biophys Res Commun, 141:904. 1986.
13.
RiyaziN., SlagboomE., de CraenA.J., MeulenbeltI., Houwing-DuistermaatJ.J., KroonH.M.et al.Association of the risk of osteoarthritis with high innate production of interleukin-1 and low innate production of interleukin-10 ex vivo, upon lipopolysaccharide stimulation. Arthritis Rheum, 52:1443. 2005.
14.
RichardsonD.W., DodgeG.R.Effects of interleukin-1beta and tumor necrosis factor-alpha on expression of matrix-related genes by cultured equine articular chondrocytes. Am J Vet Res, 61:624. 2000.
15.
GoldringM.B.Osteoarthritis and cartilage: the role of cytokines. Curr Rheumatol Rep, 2:459. 2000.
16.
LotzM., BlancoF.J., von KempisJ., DudlerJ., MaierR., VilligerP.M.et al.Cytokine regulation of chondrocyte functions. J Rheumatol Suppl, 43:104. 1995.
17.
FernandesJ.C., Martel-PelletierJ., PelletierJ.P.The role of cytokines in osteoarthritis pathophysiology. Biorheology, 39:237. 2002.
18.
López-ArmadaM.J., CaramésB., Lires-DeánM., Cillero-PastorB., Ruiz-RomeroC., GaldoF.et al.Cytokines, tumor necrosis factor-μ and interleukin-1 beta, differentially regulate apoptosis in osteoarthritis cultured human chondrocytes. Osteoarthritis Cartilage, 14:660. 2006.
19.
SunL., WangX., KaplanD.L.A 3D cartilage - Inflammatory cell culture system for the modeling of human osteoarthritis. Biomaterials, 32:5581. 2011.
20.
JonesA.R., FlanneryC.R.Bioregulation of lubricin expression by growth factors and cytokines. Eur Cell Mater, 13:40. 2007.
21.
YudohK., NguyenT., NakamuraH., Hongo-MasukoK., KatoT., NishiokaK.Potential involvement of oxidative stress in cartilage senescence and development of osteoarthritis: oxidative stress induces chondrocyte telomere instability and downregulation of chondrocyte function. Arthritis Res Ther, 7:R380. 2005.
22.
MiyakiS., NakasaT., OtsukiS., GroganS.P., HigashiyamaR., InoueA.et al.MicroRNA-140 is expressed in differentiated human articular chondrocytes and modulates interleukin-1 responses. Arthritis Rheum, 60:2723. 2009.
23.
MurphyA.R., St JohnP., KaplanD.L.Modification of silk fibroin using diazonium coupling chemistry and the effects on hMSC proliferation and differentiation. Biomaterials, 29:2829. 2008.
24.
WenkE., MurphyA.R., KaplanD.L., MeinelL., MerkleH.P., UebersaxL.The use of sulfonated silk fibroin derivatives to control binding, delivery and potency of FGF-2 in tissue regeneration. Biomaterials, 31:1403. 2010.
25.
BarberoA., GroganS., SchäferD., HebererM., Mainil-VarletP., MartinI.Age related changes in human articular chondrocyte yield, proliferation and post-expansion chondrogenic capacity. Osteoarthritis Cartilage, 12:476. 2004.
26.
SnelB., LehmannG., BorkP., HuynenM.A.STRING: a web-server to retrieve and display the repeatedly occurring neighborhood of a gene. Nucleic Acids Res, 28:3442. 2000.
27.
HuangD.W., ShermanB.T., LempickiR.A.Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res, 37:1. 2009.
28.
Ghosh. S., Parker. S.T., Wang, X., Kaplan, D.L., and Lewis, J.A. Direct-write assembly of microperiodic silk fibroin scaffolds for tissue engineering applications. Adv Func Mater, 18:1883. 2008.
29.
BarthelmesK., ReynoldsA.M., PeisachE., JonkerH.R.A., DenunzioN.J., AllenK.N.et al.Engineering encodable lanthanide-binding tags into loop regions of proteins. J Am Chem Soc, 133:808. 2011.
JeffreyJ.E., AspdenR.M.Cyclooxygenase inhibition lowers prostaglandin E2 release from articular cartilage and reduces apoptosis but not proteoglycan degradation following an impact load in vitro. Arthritis Res Ther, 9:R129. 2007.
32.
MalemudC.J., GillespieH.J.The role of apoptosis in arthritis. Curr Rheum Rev, 1:131. 2005.
33.
HaywoodL., McWilliamsD.F., PearsonC.I., GillS.E., GanesanA., WilsonD.et al.Inflammation and angiogenesis in osteoarthritis. Arthritis Rheum, 48:2173. 2003.
34.
SakkasL.I., PlatsoucasC.D.The role of T cells in the pathogenesis of osteoarthritis. Arthritis Rheum, 56:409. 2007.
35.
FosangA.J., LittleC.B.Drug insight: aggrecanases as therapeutic targets for osteoarthritis. Nat Clin Pract Rheumatol, 4:420. 2008.
36.
BondesonJ., WainwrightS., HughesC., CatersonB.The regulation of ADAMTS4 and ADAMTS5 aggrecanases in osteo-arthritis: a review. Clin Exp Rheumatol, 26:139. 2008.
37.
PetersH.C., OttoT.J., EndersJ.T., JinW., MoedB.R., ZhangZ.The protective role of the pericellular matrix in chondrocyte apoptosis. Tissue Eng Part A, 17:2017. 2011.
38.
SandyJ.D., AdamsM.E., BillinghamM.E.J., PlaasA., MuirH.In vivo and in vitro stimulation of chondrocyte biosynthetic activity in early experimental osteoarthritis. Arthritis Rheum, 27:388. 1984.
39.
LittleC.B., ChristopherB., MeekerC.T., GolubS.B., LawlorK.E., FarmerP.J.et al.Blocking aggrecanase cleavage in the aggrecan interglobular domain abrogates cartilage erosion and promotes cartilage repair. J Clin Invest, 117:1627. 2007.
40.
SantangeloK.S., PieczarkaE.M., NuovoG.J., WeisbrodeS.E., BertoneA.L.Temporal expression and tissue distribution of interleukin-1b in two strains of guinea pigs with varying propensity for spontaneous knee osteoarthritis. Osteoarthritis Cartilage, 19:439. 2011.
41.
TikuM.L., ShahR., AllisonG.T.Evidence linking chondrocyte lipid peroxidation to cartilage matrix protein degradation. Possible role in cartilage aging and the pathogenesis of osteoarthritis. J Biol Chem, 275:20069. 2000.
42.
El-BikaiR., WelmanM., MargaronY., CôtéJ.F., MacqueenL., BuschmannM.D.et al.Perturbation of adhesion molecule-mediated chondrocyte-matrix interactions by 4-hydroxynonenal binding: implication in osteoarthritis pathogenesis. Arthritis Res Ther, 12:R201. 2010.
43.
YayonA., KlagsbrunM., EskoJ.D., LederP., OrnitzD.M.Cell Surface, Heparin-like Molecules Are Required for Binding of Basic Fibroblast Growth Factor to Its High Affinity Receptor. Cell, 64:841. 1991.
44.
PrattA.B., WeberF.E., SchmoekelH.G., MuR., HubbellJ.A.Synthetic extracellular matrices for in situ tissue engineering. Biotechnology, 86:27. 2004.
45.
F MiF.L., ShyuS.S., PengC.K., WuY.B., SungH.W., WangP.S.et al.Fabrication of chondroitin sulfate–chitosan composite artificial extracellular matrix for stabilization of fibroblast growth factor. J Biomed Mater Res Part A, 76:1. 2006.
46.
ParishC.R.The role of heparan sulphate in inflammation. Nat Rev Immunol, 6:633. 2006.
47.
KenigM., Gaberc-PorekarV., FondaI., MenartV.Identification of the heparin-binding domain of TNF-alpha and its use for efficient TNF-alpha purification by heparin–Sepharose affinity chromatography. J Chromatogr B, 867:119. 2008.
48.
BegO.U., UddinM., SiddiqiA.R.Analysis of heparin-binding sites in human lipoprotein lipase using synthetic peptides. J Protein Chem, 17:807. 1998.
49.
BushP.G., HallA.C.The volume and morphology of chondrocytes within non-degenerate and degenerate human articular cartilage. Osteoarthritis Cartilage, 11:242. 2003.
50.
HollowayI., KayserM., LeeD.A., BaderD.L., BentleyG., KnightM.M.Increased presence of cells with multiple elongated processes in osteoarthritic femoral head cartilage. OsteoArthritis Cartilage, 12:17. 2004.
51.
HaudenschildD.R., ChenJ., PangN., SteklovN., GroganS.P., LotzM.K.et al.Vimentin contributes to changes in chondrocyte stiffness in osteoarthritiss. J Orthop Res, 29:20. 2011.
52.
CalamiaV., RochaB., MateosJ., Fernández-PuenteP., Ruiz-RomeroC., BlancoF.J.Metabolic labeling of chondrocytes for the quantitative analysis of the interleukin-1-beta-mediated modulation of their intracellular and extracellular proteomes. J Proteome Res, 10:3701. 2011.
53.
LambrechtS., VerbruggenG., VerdonkP.C., ElewautD., DeforceD.Differential proteome analysis of normal and osteoarthritic chondrocytes reveals distortion of vimentin network in osteoarthritis. Osteoarthritis Cartilage, 16:163. 2008.
54.
Ala-KokkoL., BaldwinC.T., MoskowitzR.W., ProckopD.J.Single base mutation in the type II procollagen gene (COL2A1) as a cause of primary osteoarthritis associated with a mild chondrodysplasia. Proc Natl Acad Sci USA, 87:6565. 1990.
55.
LorenzoP., BaylissM.T., HeinegardD.Altered patterns and synthesis of extracellular matrix macromolecules in early osteoarthritis. Matrix Biol, 23:381. 2004.
56.
ReckliesA.D., BaillargeonL., WhiteC.Regulation of cartilage oligomeric matrix protein synthesis in human synovial cells and articular chondrocytes. Arthritis Rheum, 41:997. 1998.
57.
HunterD.J., LiJ., LaValleyM., BauerD.C., NevittM., DeGrootJ.et al.Cartilage markers and their association with cartilage loss on magnetic resonance imaging in knee osteoarthritis: the Boston Osteoarthritis Knee Study. Arthritis Res Ther, 9:R108. 2007.
58.
KizawaH., KouI., IidaA., SudoA., MiyamotoY., FukudaA.et al.An aspartic acid repeat polymorphism in asporin inhibits chondrogenesis and increases susceptibility to osteoarthritis. Nat Genet, 37:138. 2005.
59.
BrewC.J., CleggP.D., Boot-HandfordR.P., AndrewJ.G., HardinghamT.Gene expression in human chondrocytes in late osteoarthritis is changed in both fibrillated and intact cartilage without evidence of generalised chondrocyte hypertrophy. Ann Rheum Dis, 69:234. 2008.
60.
BluteauG., GouttenoireJ., ConrozierT., MathieuP., VignonE., RichardM.et al.Differential gene expression analysis in a rabbit model of osteoarthritis induced by anterior cruciate ligament (ACL) section. Biorheology, 39:24. 2002.
61.
AignerT., ReichenbergerE., BertlingW., KirschT., StossH., von der MarkK.Type X collagen expression in osteoarthritic and rheumatoid articular cartilage. Virchows Arch B Cell Pathol Incl Mol Pathol, 63:205. 1993.
62.
AignerT., ZienA., GehrsitzA., GebhardP.M., McKennaL.Anabolic and catabolic gene expression pattern analysis in normal versus osteoarthritic cartilage using complementary DNA-array technology. Arthritis Rheum, 44:2777. 2001.
63.
LoeserR.F., CarlsonC.S., McGeeM.P.Expression of beta 1 integrins by cultured articular chondrocytes and in osteoarthritic cartilage. Exp Cell Res, 217:248. 1995.
64.
McCullochC.A., DowneyG.P., El-GabalawyH.Signalling platforms that modulate the inflammatory response: new targets for drug development. Nat Rev Drug Discov, 5:864. 2006.
65.
AroraP.D., MaJ., MinW., CruzT., McCullochC.A.Interleukin-1-induced calcium flux in human fibroblasts is mediated through focal adhesions. J Biol Chem, 270:6042. 1995.
66.
LoY.Y., LuoL., McCullochC.A., CruzT.F.Requirements of focal adhesions and calcium fluxes for interleukin-1-induced ERK kinase activation and c-fos expression in fibroblasts. J Biol Chem, 273:7059. 1998.
67.
MusumeciG., LoretoC., CarnazzaM.L., StrehinI., ElisseeffJ.OA cartilage derived chondrocytes encapsulated in poly(ethylene glycol) diacrylate (PEGDA) for the evaluation of cartilage restoration, apoptosis in an in vitro model. Histol Histopathol, 26:1265. 2011.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
