The aim of this study was to assess the immune modulatory properties of human mesenchymal stromal cells obtained from bone marrow (BM-MSCs), fat (ASCs), and cord blood (CB-MSCs) in the presence of a hydroxyapatite and tricalcium-phosphate (HA/TCP) biomaterial as a scaffold for MSC delivery. In resting conditions, a short-term culture with HA/TCP did not modulate the anti-apoptotic and suppressive features of the various MSC types toward T, B, and NK cells; in addition, when primed with inflammatory cytokines, MSCs similarly increased their suppressive capacities in the presence or absence of HA/TCP. The long-term culture of BM-MSCs with HA/TCP induced an osteoblast-like phenotype with upregulation of OSTERIX and OSTEOCALCIN, similar to what was obtained with dexamethasone and, to a higher extent, with bone morphogenetic protein 4 (BMP-4) treatment. MSC-derived osteoblasts did not trigger immune cell activation, but were less efficient than undifferentiated MSCs in inhibiting stimulated T and NK cells. Interestingly, their suppressive machinery included not only the activation of indoleamine-2,3 dioxygenase (IDO), which plays a central role in T-cell inhibition, but also cyclooxygenase-2 (COX-2) that was not significantly involved in the immune modulatory effect of human undifferentiated MSCs. Since COX-2 is significantly involved in bone healing, its induction by HA/TCP could also contribute to the therapeutic activity of MSCs for bone tissue engineering.
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References
1.
OkamotoK., and TakayanagiH.Regulation of bone by the adaptive immune system in arthritis. Arthritis Res Ther, 13, 219, 2011.
2.
TakayanagiH.Osteoimmunology: shared mechanisms and crosstalk between the immune and bone systems. Nat Rev Immunol, 7, 292, 2007.
3.
Schmidt-BleekK., SchellH., SchulzN., HoffP., PerkaC., ButtgereitF., VolkH-D., LienauJ., and DudaG.N.Inflammatory phase of bone healing initiates the regenerative healing cascade. Cell Tissue Res, 347, 567, 2012.
4.
ClaesL., RecknagelS., and IgnatiusA.Fracture healing under healthy and inflammatory conditions. Nat Rev Rheumatol, 8, 133, 2012.
5.
AudigéL., GriffinD., BhandariM., KellamJ., and RüediT.P.Path analysis of factors for delayed healing and nonunion in 416 operatively treated tibial shaft fractures. Clin Orthop Relat Res, 438, 221, 2005.
6.
DimitriouR., JonesE., McGonagleD., and GiannoudisP.V.Bone regeneration: current concepts and future directions. BMC Med, 9, 66, 2011.
7.
LinC.-L., FangC.-K., ChiuF.-Y., ChenC.-M., and ChenT.-H.Revision with dynamic compression plate and cancellous bone graft for aseptic nonunion after surgical treatment of humeral shaft fracture. J Trauma, 67, 1393, 2009.
8.
KurzL.T., GarfinS.R., and BoothR.E.Harvesting autogenous iliac bone grafts. A review of complications and techniques. Spine, 14, 1324, 1989.
9.
MyeroffC., and ArchdeaconM.Autogenous bone graft: donor sites and techniques. J Bone Joint Surg Am, 93, 2227, 2011.
10.
BuenoE.M., and GlowackiJ.Cell-free and cell-based approaches for bone regeneration. Nat Rev Rheumatol, 5, 685, 2009.
11.
DupontK.M., BoerckelJ.D., StevensH.Y., DiabT., KolambkarY.M., TakahataM., SchwarzE.M., and GuldbergR.E.Synthetic scaffold coating with adeno-associated virus encoding BMP2 to promote endogenous bone repair. Cell Tissue Res, 347, 575, 2012.
12.
CordonnierT., LangonnéA., SohierJ., LayrolleP., RossetP., SensébéL., and DeschaseauxF.Consistent osteoblastic differentiation of human mesenchymal stem cells with bone morphogenetic protein 4 and low serum. Tissue Eng Part C Methods, 17, 249, 2010.
13.
UccelliA., MorettaL., and PistoiaV.Mesenchymal stem cells in health and disease. Nat Rev Immunol, 8, 726, 2008.
14.
GotherstromC., RingdenO., TammikC., ZetterbergE., WestgrenM., and Le BlancK.Immunologic properties of human fetal mesenchymal stem cells. Am J Obstet Gynecol, 190, 239, 2004.
15.
DingS.J., ShieM.Y., HoshibaT., KawazoeN., ChenG., and ChangH.C.Osteogenic differentiation and immune response of human bone-marrow-derived mesenchymal stem cells on injectable calcium-silicate-based bone grafts. Tissue Eng Part A, 16, 2343, 2010.
16.
WangL., LuX.F., LuY.R., LiuJ., GaoK., ZengY.Z., LiS.F., LiY.P., ChengJ.Q., TanW.D., and WanL.Immunogenicity and immune modulation of osteogenic differentiated mesenchymal stem cells from Banna minipig inbred line. Transplant Proc, 38, 2267, 2006.
17.
Le BlancK., TammikC., RosendahlK., ZetterbergE., and RingdenO.HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells. Exp Hematol, 31, 890, 2003.
18.
Le BlancK., FrassoniF., BallL., LocatelliF., RoelofsH., LewisI., LaninoE., SundbergB., BernardoM.E., RembergerM., DiniG., EgelerR.M., BacigalupoA., FibbeW., and RingdénO.Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study. Lancet, 371, 1579, 2008.
19.
ConnickP., KolappanM., CrawleyC., WebberD.J., PataniR., MichellA.W., DuM.-Q., LuanS.-L., AltmannD.R., ThompsonA.J., CompstonA., ScottM.A., MillerD.H., and ChandranS.Autologous mesenchymal stem cells for the treatment of secondary progressive multiple sclerosis: an open-label phase 2a proof-of-concept study. Lancet Neurol, 11, 150, 2012.
20.
TanJ., WuW., XuX., LiaoL., ZhengF., MessingerS., SunX., ChenJ., YangS., CaiJ., GaoX., PileggiA., and RicordiC.Induction therapy with autologous mesenchymal stem cells in living-related kidney transplants: a randomized controlled trial. JAMA, 307, 1169, 2012.
21.
MariniJ.C., and ForlinoA.Replenishing cartilage from endogenous stem cells. N Engl J Med, 366, 2522, 2012.
22.
VacantiC.A., BonassarL.J., VacantiM.P., and ShufflebargerJ.Replacement of an avulsed phalanx with tissue-engineered bone. N Engl J Med, 344, 1511, 2001.
23.
XuJ., WangW., LudemanM., ChengK., HayamiT., LotzJ.C., and KapilaS.Chondrogenic differentiation of human mesenchymal stem cells in three-dimensional alginate gels. Tissue Eng Part A, 14, 667, 2008.
24.
CordonnierT., LayrolleP., GaillardJ., LangonnéA., SensebéL., RossetP., and SohierJ.3D environment on human mesenchymal stem cells differentiation for bone tissue engineering. J Mater Sci Mater Med, 21, 981, 2010.
25.
RochT., KrügerA., KratzK., MaN., JungF., and LendleinA.Immunological evaluation of polystyrene and poly(ether imide) cell culture inserts with different roughness. Clin Hemorheol Microcirc, 52, 375, 2012.
26.
FellahB.H., DelormeB., SohierJ., MagneD., HardouinP., and LayrolleP.Macrophage and osteoblast responses to biphasic calcium phosphate microparticles. J Biomed Mater Res A, 93, 1588, 2010.
27.
KramperaM., GalipeauJ., ShiY., TarteT., and SensebeL.Immunological characterization of multipotent mesenchymal stromal cells—The International Society for Cellular Therapy (ISCT) working proposal. Cytotherapy, 15, 1054, 2013.
BenvenutoF., FerrariS., GerdoniE., GualandiF., FrassoniF., PistoiaV., MancardiG., and UccelliA.Human mesenchymal stem cells promote survival of T cells in a quiescent state. Stem Cells, 27, 1753, 2007.
30.
Di NicolaM., Carlo-StellaC., MagniM., MilanesiM., LongoniP.D., MatteucciP., GrisantiS., and GianniA.M.Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood, 99, 3838, 2002.
31.
MenardC., PacelliL., BassiG., DulongJ., BifariF., BezierI., ZanoncelloJ., RicciardiM., LatourM., BourinP., SchrezenmeierH., SensebéL., TarteK., and KramperaM.Clinical-grade mesenchymal stromal cells produced under various GMP processes differ in their immunomodulatory properties: standardization of immune quality controls. Stem Cells Dev, 22, 1789, 2013.
32.
FeketeN., GadelorgeM., FurstD., MaurerC., DausendJ., Fleury-CappellessoS., MailanderV., LotfiR., IgnatiusA., SensebeL., BourinP., SchrezenmeierH., and RojewskiM.T.Platelet lysate from whole blood-derived pooled platelet concentrates and apheresis-derived platelet concentrates for the isolation and expansion of human bone marrow mesenchymal stromal cells: production process, content and identification of active components. Cytotherapy, 14, 540, 2012.
33.
KhanY., YaszemskiM.J., MikosA.G., and LaurencinC.T.Tissue engineering of bone: material and matrix considerations. J Bone Joint Surg Am, 90, 36, 2008.
34.
PetiteH., ViateauV., BensaïdW., MeunierA., De PollakC., BourguignonM., OudinaK., SedelL., and GuilleminG.Tissue-engineered bone regeneration. Nat Biotechnol, 18, 959, 2000.
35.
DupontK.M., SharmaK., StevensH.Y., BoerckelJ.D., GarciaA.J., and GuldbergR.E.Regenerative medicine special feature: human stem cell delivery for treatment of large segmental bone defects. PNAS, 107, 3305, 2010.
36.
GnecchiM., HeH., LiangO.D., MeloL.G., MorelloF., MuH., NoiseuxN., ZhangL., PrattR.E., IngwallJ.S., and DzauV.J.Paracrine action accounts for marked protection of ischemic heart by Akt-modified mesenchymal stem cells. Nat Med, 11, 367, 2011.
37.
AtkinsG.J., WelldonK.J., HoldingC.A., HaynesD.R., HowieD.W., and FindlayD.M.The induction of a catabolic phenotype in human primary osteoblasts and osteocytes by polyethylene particles. Biomaterials, 30, 3672, 2009.
38.
LavenusS., PiletP., GuicheuxJ., WeissP., LouarnG., and LayrolleP.Behaviour of mesenchymal stem cells, fibroblasts and osteoblasts on smooth surfaces. Acta Biomater, 30, 1525, 2011.
39.
KernS., EichlerH., StoeveJ., KlüterH., and BiebackK.Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells, 24, 1294, 2006.
40.
ZhangZ.-Y., TeohS.-H., HuiJ.H.P., FiskN.M., ChoolaniM., and ChanJ.K.Y.The potential of human fetal mesenchymal stem cells for off-the-shelf bone tissue engineering application. Biomaterials, 33, 2656, 2012.
41.
Al-NbaheenM., VishnubalajiR., AliD., BouslimiA., Al-JassirF., MeggesM., PrigioneA., AdjayeJ., KassemM., and AldahmashA.Human stromal (mesenchymal) stem cells from bone marrow, adipose tissue and skin exhibit differences in molecular phenotype and differentiation potential. Stem Cell Rev, 9, 32, 2013.
42.
LeeJ.-S., LeeJ.-M., and ImG.-I.Electroporation-mediated transfer of Runx2 and Osterix genes to enhance osteogenesis of adipose stem cells. Biomaterials, 32, 760, 2011.
43.
PinheiroC.H., de QueirozJ.C., Guimaraes-FerreiraL., VitzelK.F., NachbarR.T., de SousaL.G., de Souza-JrA.L., NunesM.T., and CuriR.Local injections of adipose-derived mesenchymal stem cells modulate inflammation and increase angiogenesis ameliorating the dystrophic phenotype in dystrophin-deficient skeletal muscle. Stem Cell Rev, 8, 363, 2012.
44.
ArinzehT.L., TranT., McalaryJ., and DaculsiG.A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. Biomaterials, 26, 3631, 2005.
45.
HabibovicP., YuanH., van der ValkC.M., MeijerG., Van BlitterswijkC.A., and de GrootK.3D microenvironment as essential element for osteoinduction by biomaterials. Biomaterials, 26, 3565, 2005.
46.
JégouxF., GoyenvalleE., CognetR., MalardO., MoreauF., DaculsiG., and AguadoE.Mandibular segmental defect regenerated with macroporous biphasic calcium phosphate, collagen membrane, and bone marrow graft in dogs. Arch Otolaryngol Head Neck Surg, 136, 971, 2010.
47.
EyckmansJ., RobertsS.J., BolanderJ., SchrootenJ., ChenC.S., and LuytenF.P.Mapping calcium phosphate activated gene networks as a strategy for targeted osteoinduction of human progenitors. Biomaterials, 34, 4612, 2013.
48.
ChaiY.C., RobertsS.J., DesmetE., KerckhofsG., van GastelN., GerisL., CarmelietG., SchrootenJ., and LuytenF.P.Mechanisms of ectopic bone formation by human osteoprogenitor cells on CaP biomaterial carriers. Biomaterials, 33, 3127, 2012.
49.
RenG., SuJ., ZhangL., ZhaoX., LingW., L'huillieA., ZhangJ., LuY., RobertsA., and JiW.Species variation in the mechanisms of mesenchymal stem cell-mediated immunosuppression. Stem Cells, 27, 1954, 2009.
50.
LangJ., KellyM., FreedB.M., McCarterM.D., KedlR.M., TorresR.M., and PelandaR.Studies of lymphocyte reconstitution in a humanized mouse model reveal a requirement of T cells for human B cell maturation. J Immunol, 190, 2090, 2013.
51.
SkjødtH., MøllerT., and FreieslebenS.F.Human osteoblast-like cells expressing MHC class II determinants stimulate allogeneic and autologous peripheral blood mononuclear cells and function as antigen-presenting cells. Immunology, 68, 416, 1989.
52.
WardW.G., GautreauxM.D., LippertD.C., and BolesC.HLA sensitization and allograft bone graft incorporation. Clin Orthop Relat Res, 466, 1837, 2008.
53.
LauK.H., KothariV., DasA., ZhangX.B., and BaylinkD.J.Cellular and molecular mechanisms of accelerated fracture healing by COX2 gene therapy: studies in a mouse model of multiple fractures. Bone, 53, 369, 2013.
54.
MartinP., and LeibovichS.J.Inflammatory cells during wound repair: the good, the bad and the ugly. Trends Cell Biol, 15, 599, 2005.
55.
HuangH., ZhaoN., XuX., XuY., LiS., ZhangJ., and YangP.Dose-specific effects of tumor necrosis factor alpha on osteogenic differentiation of mesenchymal stem cells. Cell Prolif, 44, 420, 2011.
56.
LiuY., WangL., KikuiriT., AkiyamaK., ChenC., XuX., YangR., ChenW., WangS., and ShiS.Mesenchymal stem cell-based tissue regeneration is governed by recipient T lymphocytes via IFN-γ and TNF-α. Nat Med, 17, 1594, 2011.
57.
GerstenfeldL.C., ChoT.J., KonT., AizawaT., TsayA., FitchJ., BarnesG.L., GravesD.T., and EinhornT.A.Impaired fracture healing in the absence of TNF-alpha signaling: the role of TNF-alpha in endochondral cartilage resorption. J Bone Miner Res, 18, 1584, 2003.
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