Restricted accessResearch articleFirst published online 2011-05
Osteochondral Tissue Formation Through Adipose-Derived Stromal Cell Differentiation on Biomimetic Polycaprolactone Nanofibrous Scaffolds with Graded Insulin and Beta-Glycerophosphate Concentrations
The ability to fabricate tissue engineering scaffolds containing systematic gradients in the distributions of stimulators provides additional means for the mimicking of the important gradients observed in native tissues. Here the concentration distributions of two bioactive agents were varied concomitantly for the first time (one increasing, whereas the other decreasing monotonically) in between the two sides of a nanofibrous scaffold. This was achieved via the application of a new processing method, that is, the twin-screw extrusion and electrospinning method, to generate gradients of insulin, a stimulator of chondrogenic differentiation, and β-glycerophosphate (β-GP), for mineralization. The graded poly(ɛ-caprolactone) mesh was seeded with human adipose-derived stromal cells and cultured over 8 weeks. The resulting tissue constructs were analyzed for and revealed indications of selective differentiation of human adipose-derived stromal cells toward chondrogenic lineage and mineralization as functions of position as a result of the corresponding concentrations of insulin and β-GP. Chondrogenic differentiation of the stem cells increased at insulin-rich locations and mineralization increased at β-GP-rich locations.
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References
1.
MoffatK.L., SunW.H., PenaP.E., ChahineN.O., DotyS.B., AteshianG.A., HungC.T., LuH.H.Characterization of the structure-function relationship at the ligament-to-bone interface. Proc Natl Acad Sci USA, 105:7947. 2008.
2.
McLauchlanG.J., GardnerD.L.Sacral and iliac articular cartilage thickness and cellularity: relationship to subchondral bone end-plate thickness and cancellous bone density. Rheumatology, 41:375. 2002.
3.
PhillipsJ.E., BurnsK.L., Le DouxJ.M., GuldbergR.E., GarciaA.J.Engineering graded tissue interfaces. Proc Natl Acad Sci USA, 105:12170. 2008.
4.
BradleyD.A., MuthuveluP., EllisR.E., GreenE.M., AttenburrowD., BarrettR., ArkillK., ColridgeD.B., WinloveC.P.Characterisation of mineralisation of bone and cartilage: X-ray diffraction and Ca and SrK alpha X-ray fluorescence microscopy. Nucl Instrum Meth B, 263:1. 2007.
5.
JiangJ., NicollS.B., LuH.H.Co-culture of osteoblasts and chondrocytes modulates cellular differentiation in vitro. Biochem Biophys Res Commun, 338:762. 2005.
6.
SarazinP., RoyX., FavisB.D.Controlled preparation and properties of porous poly(L-lactide) obtained from a co-continuous blend of two biodegradable polymers. Biomaterials, 25:5965. 2004.
7.
OzkanS., KalyonD.M., YuX., McKelveyC., LowingerM.Multifunctional polycaprolactone (PCL) scaffolds integrated for controlled release and tissue engineering: In vitro evaluation of released protein secondary structure stability, release profile and biocompatibility. Biomaterials, 30:4336. 2009.
8.
LuoY., KoblerJ.B., ZeitelsS.M., LangerR.Effects of growth factors on extracellular matrix production by vocal fold fibroblasts in 3-dimensional culture. Tissue Eng, 12:3365. 2006.
UebersaxL., MerkleH.P., MeinelL.Insulin-like growth factor I releasing silk fibroin scaffolds induce chondrogenic differentiation of human mesenchymal stem cells. J Control Release, 127:12. 2008.
11.
LiC., VepariC., JinH.J., KimH.J., KaplanD.L.Electrospun silk-BMP-2 scaffolds for bone tissue engineering. Biomaterials, 27:3115. 2006.
12.
LevenbergS., HuangN.F., LavikE., RogersA.B., Itskovitz-EldorJ., LangerR.Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds. Proc Natl Acad Sci USA, 100:12741. 2003.
13.
WangX., WenkE., ZhangX., MeinelL., Vunjak-NovakovicG., KaplanD.L.Growth factor gradients via microsphere delivery in biopolymer scaffolds for osteochondral tissue engineering. J Control Release, 134:81. 2009.
14.
BenoitD.S., SchwartzM.P., DurneyA.R., AnsethK.S.Small functional groups for controlled differentiation of hydrogel-encapsulated human mesenchymal stem cells. Nat Mater, 7:816. 2008.
EriskenC., KalyonD.M., WangH.J.A hybrid twin screw extrusion/electrospinning method to process nanoparticle-incorporated electrospun nanofibres. Nanotechnology, 19:165302. 2008.
17.
EriskenC., KalyonD.M., WangH.Functionally graded electrospun polycaprolactone and beta-tricalcium phosphate nanocomposites for tissue engineering applications. Biomaterials, 29:4065. 2008.
18.
ZukP.A., ZhuM., MizunoH., HuangJ., FutrellJ.W., KatzA.J., BenhaimP., LorenzH.P., HedrickM.H.Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng, 7:211. 2001.
19.
HadhazyC., DedukhN.V.[Effect of insulin on cartilage differentiation in vitro]Biull Eksp Biol Med, 105:219. 1988.
20.
JulianD., AbbottU.K.An avian model for comparative studies of insulin teratogenicity. Anat Histol Embryol, 27:313. 1998.
21.
MaroltD., AugstA., FreedL.E., VepariC., FajardoR., PatelN., GrayM., FarleyM., KaplanD., Vunjak-NovakovicG.Bone and cartilage tissue constructs grown using human bone marrow stromal cells, silk scaffolds and rotating bioreactors. Biomaterials, 27:6138. 2006.
22.
MaorG., SilbermannM., von derM.K., HeingardD., LaronZ.Insulin enhances the growth of cartilage in organ and tissue cultures of mouse neonatal mandibular condyle. Calcif Tissue Int, 52:291. 1993.
23.
AllanK.S., PilliarR.M., WangJ., GrynpasM.D., KandelR.A.Formation of biphasic constructs containing cartilage with a calcified zone interface. Tissue Eng, 13:167. 2007.
24.
ChungC.H., GolubE.E., ForbesE., TokuokaT., ShapiroI.M.Mechanism of action of beta-glycerophosphate on bone cell mineralization. Calcif Tissue Int, 51:305. 1992.
25.
TenenbaumH.C.Role of organic phosphate in mineralization of bone in vitro. J Dent Res, 60,Spec No C:1586. 1981.
26.
WallM.E., RachlinA., OteyC.A., LoboaE.G.Human adipose-derived adult stem cells upregulate palladin during osteogenesis and in response to cyclic tensile strain. Am J Physiol Cell Physiol, 293:C1532. 2007.
27.
WuY.N., YangZ., HuiJ.H., OuyangH.W., LeeE.H.Cartilaginous ECM component-modification of the micro-bead culture system for chondrogenic differentiation of mesenchymal stem cells. Biomaterials, 28:4056. 2007.
28.
EriskenC., KalyonD.M., WangH.Viscoelastic and biomechanical properties of osteochondral tissue constructs generated from graded polycaprolactone and beta-tricalcium phosphate composites. J Biomech Eng, 132:091013. 2010.
29.
LemkeM.J., ChurchillP.F., WetzelR.G.Effect of substrate and cell surface hydrophobicity on phosphate utilization in bacteria. Appl Environ Microbiol, 61:913. 1995.
30.
KondoS., XieW.-F., ChaS.H., SandellL.J.Discovery and characteristic features of cartilage-derived retinoic acid-sensitive protein (CD-RAP)Eur Cells Mater, 8:57. 1998.
31.
SakanoS., ZhuY., SandellL.J.Cartilage-derived retinoic acid-sensitive protein and type II collagen expression during fracture healing are potential targets for Sox9 regulation. J Bone Miner Res, 14:1891. 1999.
32.
DietzU.H., SandellL.J.Cloning of a retinoic acid-sensitive mRNA expressed in cartilage and during chondrogenesis. J Biol Chem, 271:3311. 1996.
33.
BosserhoffA.K., BuettnerR.Establishing the protein MIA (melanoma inhibitory activity) as a marker for chondrocyte differentiation. Biomaterials, 24:3229. 2003.
34.
Dell'AccioF., De BariC., LuytenF.P.Molecular markers predictive of the capacity of expanded human articular chondrocytes to form stable cartilage in vivo. Arthritis Rheum, 44:1608. 2001.
35.
DengC., Wynshaw-BorisA., ZhouF., KuoA., LederP.Fibroblast growth factor receptor 3 is a negative regulator of bone growth. Cell, 84:911. 1996.
36.
ColvinJ.S., BohneB.A., HardingG.W., McEwenD.G., OrnitzD.M.Skeletal overgrowth and deafness in mice lacking fibroblast growth factor receptor 3. Nat Genet, 12:390. 1996.
37.
SuN., YangJ., XieY., DuX., LuX., YinZ., YinL., QiH., ZhaoL., FengJ., ChenL.Gain-of-function mutation of FGFR3 results in impaired fracture healing due to inhibition of chondrocyte differentiation. Biochem Biophys Res Commun, 376:454. 2008.
38.
ZuscikM.J., HiltonM.J., ZhangX., ChenD., O'KeefeR.J.Regulation of chondrogenesis and chondrocyte differentiation by stress. J Clin Invest, 118:429. 2008.
39.
GhoshP., SmithM.Osteoarthritis, genetic and molecular mechanisms. Biogerontology, 3:85. 2002.
40.
LefebvreV., Peeters-JorisC., VaesG.Modulation by interleukin 1 and tumor necrosis factor alpha of production of collagenase, tissue inhibitor of metalloproteinases and collagen types in differentiated and dedifferentiated articular chondrocytes. Biochim Biophys Acta, 1052:366. 1990.
41.
BenyaP.D., PadillaS.R., NimniM.E.The progeny of rabbit articular chondrocytes synthesize collagen types I and III and type I trimer, but not type II. Verifications by cyanogen bromide peptide analysis. Biochemistry, 16:865. 1977.
42.
YoonY.M., KimS.J., OhC.D., JuJ.W., SongW.K., YooY.J., HuhT.L., ChunJ.S.Maintenance of differentiated phenotype of articular chondrocytes by protein kinase C and extracellular signal-regulated protein kinase. J Biol Chem, 277:8412. 2002.
43.
BonaventureJ., KadhomN., Cohen-SolalL., NgK.H., BourguignonJ., LasselinC., FreisingerP.Reexpression of cartilage-specific genes by dedifferentiated human articular chondrocytes cultured in alginate beads. Exp Cell Res, 212:97. 1994.
44.
WoodfieldT.B., van BlitterswijkC.A., DeW.J., SimsT.J., HollanderA.P., RiesleJ.Polymer scaffolds fabricated with pore-size gradients as a model for studying the zonal organization within tissue-engineered cartilage constructs. Tissue Eng, 11:1297. 2005.
NgK.W., AteshianG.A., HungC.T.Zonal Chondrocytes seeded in a layered agarose hydrogel create engineered cartilage with depth-dependent cellular and mechanical inhomogeneity. Tissue Eng Part A, 15:2315. 2009.
49.
SinghM., MorrisC.P., EllisR.J., DetamoreM.S., BerklandC.Microsphere-based seamless scaffolds containing macroscopic gradients of encapsulated factors for tissue engineering. Tissue Eng Part C Methods, 14:299. 2008.
50.
ZeltingerJ., SherwoodJ.K., GrahamD.A., MuellerR., GriffithL.G.Effect of pore size and void fraction on cellular adhesion, proliferation, and matrix deposition. Tissue Eng, 7:557. 2001.
51.
LiX.R., XieJ.W., LipnerJ., YuanX.Y., ThomopoulosS., XiaY.N.Nanofiber scaffolds with gradations in mineral content for mimicking the tendon-to-bone insertion site. Nano Lett, 9:2763. 2009.
52.
ChangY.L., StanfordC.M., KellerJ.C.Calcium and phosphate supplementation promotes bone cell mineralization: implications for hydroxyapatite (HA)-enhanced bone formation. J Biomed Mater Res, 52:270. 2000.
53.
YuH., GraingerD.W.Modified release of hydrophilic, hydrophobic and peptide agents from ionized amphiphilic gel networks. J Control Release, 34:117. 1995.
54.
SerravezzaJ.C., EndoF., DehaanR.L., ElsasL.J.Insulin-induced receptor loss reduces responsiveness of chick heart-cells to insulin. Endocrinology, 113:497. 1983.
55.
BhattaraiN., EdmondsonD., VeisehO., MatsenF.A., ZhangM.Q.Electrospun chitosan-based nanofibers and their cellular compatibility. Biomaterials, 26:6176. 2005.
56.
KempS.F., MutchnickM., HintzR.L.Hormonal-control of protein-synthesis in chick chondrocytes—a comparison of effects of insulin, somatomedin-c and triiodothyronine. Acta Endocrinol, 107:179. 1984.
57.
MuramatsuT., KitaK., NakagawaS., OkumuraJ.Effects of insulin, growth-hormone, IGF-I and IGF-II on phenylalanine extraction in the chicken-embryo cultured in vitro. Comp Biochem Physiol A Physiol, 104:507. 1993.
58.
ScottJ.E., DorlingJ.Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemie, 5:221. 1965.
59.
ZhuW., MowV.C., KoobT.J., EyreD.R.Viscoelastic shear properties of articular cartilage and the effects of glycosidase treatments. J Orthop Res, 11:771. 1993.
NeumannK., DehneT., EndresM., ErggeletC., KapsC., RingeJ., SittingerM.Chondrogenic differentiation capacity of human mesenchymal progenitor cells derived from subchondral cortico-spongious bone. J Orthop Res, 26:1449. 2008.
62.
JunkerJ.P.E., SommarP., SkogM., JohnsonH., KratzG.Adipogenic, chondrogenic and osteogenic differentiation of clonally derived human dermal fibroblasts. Cells Tissues Organs, 191:105. 2010.
63.
WilliamsR., KhanI.M., RichardsonK., NelsonL., McCarthyH.E., AnalbelsiT., SinghraoS.K., DowthwaiteG.P., JonesR.E., BairdD.M., LewisH., RobertsS., ShawH.M., DudhiaJ., FaircloughJ., BriggsT., ArcherC.W.Identification and clonal characterisation of a progenitor cell sub-population in normal human articular cartilage. PLoS ONE, 5:e13246. 2010.
64.
GarretaE., GenoveE., BorrosS., SeminoC.E.Osteogenic differentiation of mouse embryonic stem cells and mouse embryonic fibroblasts in a three-dimensional self-assembling peptide scaffold. Tissue Eng, 12:2215. 2006.
65.
KnabeC., KraskaB., KochC., GrossU., ZreiqatH., StillerM.A method for immunohistochemical detection of osteogenic markers in undecalcified bone sections. Biotech Histochem, 81:31. 2006.
66.
SchutzeN., NothU., SchneidereitJ., HendrichC., JakobF.Differential expression of CCN-family members in primary human bone marrow-derived mesenchymal stem cells during osteogenic, chondrogenic and adipogenic differentiation. Cell Commun Signal, 3:5. 2005.
67.
Sophia FoxA.J., BediA., RodeoS.A.The basic science of articular cartilage: structure, composition, and function. Sports Health: A Multidisciplinary Approach, 1:461. 2009.
68.
BarshG.S., DavidK.E., ByersP.H.Type I osteogenesis imperfecta: a nonfunctional allele for pro alpha 1 (I) chains of type I procollagen. Proc Natl Acad Sci USA, 79:3838. 1982.
69.
KirschE., KriegT., NerlichA., RembergerK., MeineckeP., KunzeD., MullerP.K.Compositional analysis of collagen from patients with diverse forms of osteogenesis imperfecta. Calcif Tissue Int, 41:11. 1987.
70.
JunqueiraL.C., BignolasG., BrentaniR.R.Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J, 11:447. 1979.
71.
ZukP.A., ZhuM., AshjianP., De UgarteD.A., HuangJ.I., MizunoH., AlfonsoZ.C., FraserJ.K., BenhaimP., HedrickM.H.Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell, 13:4279. 2002.
72.
AwadH.A., WickhamM.Q., LeddyH.A., GimbleJ.M., GuilakF.Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds. Biomaterials, 25:3211. 2004.
73.
TappH., DeepeR., IngramJ.A., KuremskyM., HanleyE.N.Jr., GruberH.E.Adipose-derived mesenchymal stem cells from the sand rat: transforming growth factor beta and 3D co-culture with human disc cells stimulate proteoglycan and collagen type I rich extracellular matrix. Arthritis Res Ther, 10:R89. 2008.
74.
MusselmannK., KaneB., AlexandrouB., HassellJ.R.Stimulation of collagen synthesis by insulin and proteoglycan accumulation by ascorbate in bovine keratocytes in vitro. Invest Ophthalmol Vis Sci, 47:5260. 2006.
75.
BosserhoffA.K., KondoS., MoserM., DietzU.H., CopelandN.G., GilbertD.J., JenkinsN.A., BuettnerR., SandellL.J.Mouse CD-RAP/MIA gene: structure, chromosomal localization, and expression in cartilage and chondrosarcoma. Dev Dyn, 208:516. 1997.
76.
GubaM., BosserhoffA.K., SteinbauerM., AbelsC., AnthuberM., BuettnerR., JauchK.W.Overexpression of melanoma inhibitory activity (MIA) enhances extravasation and metastasis of A-mel 3 melanoma cells in vivo. Br J Cancer, 83:1216. 2000.
Valverde-FrancoG., BinetteJ.S., LiW., WangH., ChaiS., LaflammeF., Tran-KhanhN., QuennevilleE., MeijersT., PooleA.R., MortJ.S., BuschmannM.D., HendersonJ.E.Defects in articular cartilage metabolism and early arthritis in fibroblast growth factor receptor 3 deficient mice. Hum Mol Genet, 15:1783. 2006.
79.
ValcourtU., GouttenoireJ., MoustakasA., HerbageD., Mallein-GerinF.Functions of transforming growth factor-beta family type I receptors and smad proteins in the hypertrophic maturation and osteoblastic differentiation of chondrocytes. J Biol Chem, 277:33545. 2002.
80.
TenenbaumH.C.Levamisole and inorganic pyrophosphate inhibit beta-glycerophosphate induced mineralization of bone formed in vitro. Bone Miner, 3:13. 1987.
81.
GoughJ.E., JonesJ.R., HenchL.L.Nodule formation and mineralisation of human primary osteoblasts cultured on a porous bioactive glass scaffold. Biomaterials, 25:2039. 2004.
82.
FujitaT., MeguroT., IzumoN., YasutomiC., FukuyamaR., NakamutaH., KoidaM.Phosphate stimulates differentiation and mineralization of the chondroprogenitor clone ATDC5. Jpn J Pharmacol, 85:278. 2001.
83.
KandelR.A., BoyleJ., GibsonG., CruzT., SpeagleM.In vitro formation of mineralized cartilagenous tissue by articular chondrocytes. In Vitro Cell Dev Biol Anim, 33:174. 1997.
84.
WangC.C., HungC.T., MowV.C.An analysis of the effects of depth-dependent aggregate modulus on articular cartilage stress-relaxation behavior in compression. J Biomech, 34:75. 2001.
85.
ArmstrongC.G., LaiW.M., MowV.C.An analysis of the unconfined compression of articular cartilage. J Biomech Eng, 106:165. 1984.
86.
ParkS., HungC.T., AteshianG.A.Mechanical response of bovine articular cartilage under dynamic unconfined compression loading at physiological stress levels. Osteoarthritis Cartilage, 12:65. 2004.
87.
GarneroP., BorelO., GineytsE., DuboeufF., SolbergH., BouxseinM.L., ChristiansenC., DelmasP.D.Extracellular post-translational modifications of collagen are major determinants of biomechanical properties of fetal bovine cortical bone. Bone, 38:300. 2006.
88.
BurrD.B.The contribution of the organic matrix to bone's material properties. Bone, 31:8. 2002.
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