To investigate the effect of soluble factors released from human nasal chondrocytes (NCs) on cocultured human bone marrow mesenchymal stem cells (MSCs) and NC tissue-engineered constructs. Cartilage engineered from pure NCs on a three-dimensional (3D) porous collagen scaffold was cultured indirectly in a Transwell system with cartilage engineered from a direct coculture of human bone marrow-derived MSCs and NCs on a 3D porous collagen scaffold. The soluble factors were measured in the conditioned media from the different chambers of the Transwell system. Engineered cartilage from cocultures exposed to the pure NC construct exhibited reduced chondrogenic potential relative to control constructs, shown by reduced extracellular matrix deposition and increased expression of hypertrophic markers. Analysis of the soluble factors within the conditioned media showed an increase in inflammatory cytokines in the coculture chamber exposed to the pure NC construct. Principal component analysis revealed that the majority of the data variance could be explained by proinflammatory factors and hypertrophic chondrogenesis. In conclusion, our data suggest that inflammatory cytokines derived from NCs reduce the chondrogenic potential of coculture engineered cartilage through the induction of hypertrophic chondrogenesis.
Impact statement
The use of engineered cartilage from cocultured nasal chondrocytes (NCs) and mesenchymal stem cells for nasal cartilage reconstruction may be problematic. Our data suggest that the soluble factors from surrounding native NCs in the cartilage to be fixed can compromise the quality of the engineered cartilage if used in reconstructive surgery.
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References
1.
FulcoI., MiotS., HaugM.D., BarberoA., WixmertenA., FelicianoS., WolfF., JundtG., MarsanoA., FarhadiJ., HebererM., JakobM., SchaeferD.J., and MartinI.Engineered autologous cartilage tissue for nasal reconstruction after tumour resection: an observational first-in-human trial. Lancet, 384, 337, 2014.
2.
MummeM., BarberoA., MiotS., WixmertenA., FelicianoS., WolfF., AsnaghiA.M., BaumhoerD., BieriO., KretzschmarM., PagenstertG., HaugM., SchaeferD.J., MartinI., and JakobM.Nasal chondrocyte-based engineered autologous cartilage tissue for repair of articular cartilage defects: an observational first-in-human trial. Lancet, 388, 1985, 2016.
3.
PelttariK., MummeM., BarberoA., and MartinI.Nasal chondrocytes as a neural crest-derived cell source for regenerative medicine. Curr Opin Biotechnol, 47, 1, 2017.
4.
PelttariK., PippengerB., MummeM., FelicianoS., ScottiC., Mainil-VarletP., ProcinoA., von RechenbergB., SchwambornT., JakobM., CilloC., BarberoA., and MartinI.Adult human neural crest-derived cells for articular cartilage repair. Sci Transl Med, 6, 251ra119, 2014.
5.
AndrewsS.H.J., KunzeM., Mulet-SierraA., WilliamsL., AnsariK., OsswaldM., and AdesidaA.B.Strategies to mitigate variability in engineering human nasal cartilage. Sci Rep, 7, 6490, 2017.
6.
TayA.G., FarhadiJ., SuetterlinR., PiererG., HebererM., and MartinI.Cell yield, proliferation, and postexpansion differentiation capacity of human ear, nasal, and rib chondrocytes. Tissue Eng, 10, 762, 2004.
7.
CandrianC., VonwilD., BarberoA., BonacinaE., MiotS., FarhadiJ., WirzD., DickinsonS., HollanderA., JakobM., LiZ., AliniM., HebererM., and MartinI.Engineered cartilage generated by nasal chondrocytes is responsive to physical forces resembling joint loading. Arthritis Rheum, 58, 197, 2007.
8.
KafienahW., JakobM., DemarteauO., FrazerA., BarkerM.D., MartinI., and HollanderA.P.Three-dimensional tissue engineering of hyaline cartilage: comparison of adult nasal and articular chondrocytes. Tissue Eng, 8, 817, 2002.
9.
VedicherlaS., and BuckleyC.T.In vitro extracellular matrix accumulation of nasal and articular chondrocytes for intervertebral disc repair. Tissue Cell, 49, 503, 2017.
10.
GayM.H., MehrkensA., RittmannM., HaugM., BarberoA., MartinI., and SchaerenS.Nose to back: compatibility of nasal chondrocytes with environmental conditions mimicking a degenerated intervertebral disc. Eur Cell Mater, 37, 214, 2019.
PittengerM.F., MackayA.M., BeckS.C., JaiswalR.K., DouglasR., MoscaJ.D., MoormanM.A., SimonettiD.W., CraigS., and MarshakD.R.Multilineage potential of adult human mesenchymal stem cells. Science, 284, 143, 1999.
13.
MackayA.M., BeckS.C., MurphyJ.M., BarryF.P., ChichesterC.O., and PittengerM.F.Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng, 4, 415, 1998.
14.
LeeR.H., KimB., ChoiI., KimH., ChoiH.S., SuhK., BaeY.C., and JungJ.S.Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue. Cell Physiol Biochem, 14, 311, 2004.
15.
GuoX., DuJ., ZhengQ., YangS., LiuY., DuanD., and YiC.Expression of transforming growth factor beta 1 in mesenchymal stem cells: potential utility in molecular tissue engineering for osteochondral repair. J Huazhong Univ Sci Technol Med Sci, 22, 112, 2002.
16.
BanfiA., MuragliaA., DozinB., MastrogiacomoM., CanceddaR., and QuartoR.Proliferation kinetics and differentiation potential of ex vivo expanded human bone marrow stromal cells: implications for their use in cell therapy. Exp Hematol, 28, 707, 2000.
17.
MuragliaA., CanceddaR., and QuartoR.Clonal mesenchymal progenitors from human bone marrow differentiate in vitro according to a hierarchical model. J Cell Sci, 113 (Pt 7), 1161, 2000.
18.
PelttariK., WinterA., SteckE., GoetzkeK., HennigT., OchsB.G., AignerT., and RichterW.Premature induction of hypertrophy during in vitro chondrogenesis of human mesenchymal stem cells correlates with calcification and vascular invasion after ectopic transplantation in SCID mice. Arthritis Rheum, 54, 3254, 2006.
19.
PaschosN.K., BrownW.E., EswaramoorthyR., HuJ.C., and AthanasiouK.A.Advances in tissue engineering through stem cell-based co-culture. J Tissue Eng Regen Med, 9, 488, 2015.
20.
AcharyaC., AdesidaA., ZajacP., MummeM., RiesleJ., MartinI., and BarberoA.Enhanced chondrocyte proliferation and mesenchymal stromal cells chondrogenesis in coculture pellets mediate improved cartilage formation. J Cell Physiol, 227, 88, 2012.
21.
WuL., PrinsH.J., HelderM.N., van BlitterswijkC.A., and KarperienM.Trophic effects of mesenchymal stem cells in chondrocyte co-cultures are independent of culture conditions and cell sources. Tissue Eng Part A, 18, 1542, 2012.
22.
SobacchiC., PalaganoE., VillaA., and MenaleC.Soluble factors on stage to direct mesenchymal stem cells fate. Front Bioeng Biotechnol, 5, 32, 2017.
23.
LeeC.S., BurnsedO.A., RaghuramV., KalisvaartJ., BoyanB.D., and SchwartzZ.Adipose stem cells can secrete angiogenic factors that inhibit hyaline cartilage regeneration. Stem Cell Res Ther, 3, 35, 2012.
24.
AhmedN., DreierR., GopferichA., GrifkaJ., and GrasselS.Soluble signalling factors derived from differentiated cartilage tissue affect chondrogenic differentiation of rat adult marrow stromal cells. Cell Physiol Biochem, 20, 665, 2007.
25.
WuL., LeijtenJ.C., GeorgiN., PostJ.N., van BlitterswijkC., and KarperienM.Trophic effects of mesenchymal stem cells increase chondrocyte proliferation and matrix formation. Tissue Eng Part A, 17, 1425, 2011.
26.
HwangN.S., VargheseS., PuleoC., ZhangZ., and ElisseeffJ.Morphogenetic signals from chondrocytes promote chondrogenic and osteogenic differentiation of mesenchymal stem cells. J Cell Physiol, 212, 281, 2007.
27.
TaylorD.W., AhmedN., GanL., GrossA.E., and KandelR.A.Proteoglycan and collagen accumulation by passaged chondrocytes can be enhanced through side-by-side culture with primary chondrocytes. Tissue Eng Part A, 16, 643, 2010.
28.
VatsA., BielbyR.C., TolleyN., DickinsonS.C., BoccacciniA.R., HollanderA.P., BishopA.E., and PolakJ.M.Chondrogenic differentiation of human embryonic stem cells: the effect of the micro-environment. Tissue Eng, 12, 1687, 2006.
29.
BornesT.D., JomhaN.M., Mulet-SierraA., and AdesidaA.B.Optimal seeding densities for in vitro chondrogenesis of two- and three-dimensional-isolated and -expanded bone marrow-derived mesenchymal stromal stem cells within a porous collagen scaffold. Tissue Eng Part C Methods, 22, 208, 2016.
30.
MatthiesN.F., Mulet-SierraA., JomhaN.M., and AdesidaA.B.Matrix formation is enhanced in co-cultures of human meniscus cells with bone marrow stromal cells. J Tissue Eng Regen Med, 7, 965, 2013.
31.
de WindtT.S., HendriksJ.A., ZhaoX., VonkL.A., CreemersL.B., DhertW.J., RandolphM.A., and SarisD.B.Concise review: unraveling stem cell cocultures in regenerative medicine: which cell interactions steer cartilage regeneration and how? Stem Cells Transl Med, 3, 723, 2014.
32.
GroganS.P., BarberoA., WinkelmannV., RieserF., FitzsimmonsJ.S., O'DriscollS., MartinI., and Mainil-VarletP.Visual histological grading system for the evaluation of in vitro-generated neocartilage. Tissue Eng, 12, 2141, 2006.
33.
VandesompeleJ., De PreterK., PattynF., PoppeB., Van RoyN., De PaepeA., and SpelemanF.Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol, 3, Research0034, 2002.
34.
LivakK.J., and SchmittgenT.D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25, 402, 2001.
35.
SchmittgenT.D., and LivakK.J.Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc, 3, 1101, 2008.
36.
RosenbergL.Chemical basis for the histological use of safranin O in the study of articular cartilage. J Bone Joint Surg Am, 53, 69, 1971.
37.
KianiC., ChenL., WuY.J., YeeA.J., and YangB.B.Structure and function of aggrecan. Cell Res, 12, 19, 2002.
38.
MarkwayB.D., ChoH., and JohnstoneB.Hypoxia promotes redifferentiation and suppresses markers of hypertrophy and degeneration in both healthy and osteoarthritic chondrocytes. Arthritis Res Ther, 15, R92, 2013.
39.
AdesidaA.B., Mulet-SierraA., and JomhaN.M.Hypoxia mediated isolation and expansion enhances the chondrogenic capacity of bone marrow mesenchymal stromal cells. Stem Cell Res Ther, 3, 9, 2012.
40.
BroweD.C., ColemanC.M., BarryF.P., and EllimanS.J.Hypoxia Activates the PTHrP -MEF2C Pathway to Attenuate Hypertrophy in Mesenchymal Stem Cell Derived Cartilage. Sci Rep, 9, 13274, 2019.
41.
D'AngeloM., YanZ., NooreyazdanM., PacificiM., SarmentD.S., BillingsP.C., and LeboyP.S.MMP-13 is induced during chondrocyte hypertrophy. J Cell Biochem, 77, 678, 2000.
42.
LeemanM.F., CurranS., and MurrayG.I.The structure, regulation, and function of human matrix metalloproteinase-13. Crit Rev Biochem Mol Biol, 37, 149, 2002.
43.
von der MarkK., FrischholzS., AignerT., BeierF., BelkeJ., ErdmannS., and BurkhardtH.Upregulation of type X collagen expression in osteoarthritic cartilage. Acta Orthop Scand Suppl, 266, 125, 1995.
44.
GannonJ.M., WalkerG., FischerM., CarpenterR., ThompsonR.C.Jr., and OegemaT.R.Jr. Localization of type X collagen in canine growth plate and adult canine articular cartilage. J Orthop Res, 9, 485, 1991.
45.
GryM., RiminiR., StrombergS., AsplundA., PontenF., UhlenM., and NilssonP.Correlations between RNA and protein expression profiles in 23 human cell lines. BMC Genomics, 10, 365, 2009.
46.
CapsoniF., OngariA.M., LonatiC., AccettaR., GattiS., and CataniaA.alpha-Melanocyte-stimulating-hormone (alpha-MSH) modulates human chondrocyte activation induced by proinflammatory cytokines. BMC Musculoskelet Disord, 16, 154, 2015.
47.
EmansP.J., SpaapenF., SurtelD.A., ReillyK.M., CremersA., van RhijnL.W., BulstraS.K., VonckenJ.W., and KuijerR.A novel in vivo model to study endochondral bone formation; HIF-1alpha activation and BMP expression. Bone, 40, 409, 2007.
48.
FukuiN., IkedaY., OhnukiT., HikitaA., TanakaS., YamaneS., SuzukiR., SandellL.J., and OchiT.Pro-inflammatory cytokine tumor necrosis factor-alpha induces bone morphogenetic protein-2 in chondrocytes via mRNA stabilization and transcriptional up-regulation. J Biol Chem, 281, 27229, 2006.
49.
KomuroH., OleeT., KuhnK., QuachJ., BrinsonD.C., ShikhmanA., ValbrachtJ., Creighton-AchermannL., and LotzM.The osteoprotegerin/receptor activator of nuclear factor kappaB/receptor activator of nuclear factor kappaB ligand system in cartilage. Arthritis Rheum, 44, 2768, 2001.
50.
ShimizuS., AsouY., ItohS., ChungU.I., KawaguchiH., ShinomiyaK., and MunetaT.Prevention of cartilage destruction with intraarticular osteoclastogenesis inhibitory factor/osteoprotegerin in a murine model of osteoarthritis. Arthritis Rheum, 56, 3358, 2007.
51.
HanB.K., KuzinI., GaughanJ.P., OlsenN.J., and BottaroA.Baseline CXCL10 and CXCL13 levels are predictive biomarkers for tumor necrosis factor inhibitor therapy in patients with moderate to severe rheumatoid arthritis: a pilot, prospective study. Arthritis Res Ther, 18, 93, 2016.
52.
AllamS.I., SallamR.A., ElghannamD.M., and El-GhaweetA.I.Clinical significance of serum B cell chemokine (CXCL13) in early rheumatoid arthritis patients. Egypt Rheumatol, 41, 11, 2019.
53.
De CeuninckF., CaliezA., DassencourtL., AnractP., and RenardP.Pharmacological disruption of insulin-like growth factor 1 binding to IGF-binding proteins restores anabolic responses in human osteoarthritic chondrocytes. Arthritis Res Ther, 6, R393, 2004.
54.
GoldringM.B., and BerenbaumF.The regulation of chondrocyte function by proinflammatory mediators: prostaglandins and nitric oxide. Clin Orthop Relat Res, 427Suppl, S37, 2004.
55.
KobayashiM., SquiresG.R., MousaA., TanzerM., ZukorD.J., AntoniouJ., FeigeU., and PooleA.R.Role of interleukin-1 and tumor necrosis factor alpha in matrix degradation of human osteoarthritic cartilage. Arthritis Rheum, 52, 128, 2005.
56.
FuZ., SongX., GuoL., YangL., and ChenC.Effects of conditioned medium from osteoarthritic cartilage fragments on donor-matched infrapatellar fat pad-derived mesenchymal stromal cells. Am J Sports Med, 47, 2927, 2019.
57.
ArendW.P., and DayerJ.M.Cytokines and cytokine inhibitors or antagonists in rheumatoid arthritis. Arthritis Rheum, 33, 305, 1990.
58.
KraneS.M., ConcaW., StephensonM.L., AmentoE.P., and GoldringM.B.Mechanisms of matrix degradation in rheumatoid arthritis. Ann N Y Acad Sci, 580, 340, 1990.
59.
TylerJ.A., and BentonH.P.Synthesis of type II collagen is decreased in cartilage cultured with interleukin 1 while the rate of intracellular degradation remains unchanged. Coll Relat Res, 8, 393, 1988.
60.
GoldringM.B., BirkheadJ., SandellL.J., KimuraT., and KraneS.M.Interleukin 1 suppresses expression of cartilage-specific types II and IX collagens and increases types I and III collagens in human chondrocytes. J Clin Invest, 82, 2026, 1988.
61.
HendersonB., and PettipherE.R.Arthritogenic actions of recombinant IL-1 and tumour necrosis factor alpha in the rabbit: evidence for synergistic interactions between cytokines in vivo. Clin Exp Immunol, 75, 306, 1989.
62.
SaklatvalaJ.Tumour necrosis factor alpha stimulates resorption and inhibits synthesis of proteoglycan in cartilage. Nature, 322, 547, 1986.
63.
EngvallI.L., ElkanA.C., TengstrandB., CederholmT., BrismarK., and HafstromI.Cachexia in rheumatoid arthritis is associated with inflammatory activity, physical disability, and low bioavailable insulin-like growth factor. Scand J Rheumatol, 37, 321, 2008.
64.
NeidelJ.Changes in systemic levels of insulin-like growth factors and their binding proteins in patients with rheumatoid arthritis. Clin Exp Rheumatol, 19, 81, 2001.
65.
AbruzzoJ.L.Rheumatoid arthritis: reflections on etiology and pathogenesis. Arch Phys Med Rehabil, 52, 30, 1971.
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