Many cell-based tissue-engineered cartilaginous constructs are mechanically softer than native tissue and have low content and abnormal proportions of extracellular matrix (ECM) constituents. We hypothesized that the load-bearing mechanical properties of cartilaginous constructs improve with the inclusion of collagen (COL) and proteoglycan (PG) during assembly. The objectives of this work were to determine (1) the effect of addition of PG, COL, or COL+PG on compressive properties of 2% agarose constructs and (2) the ability of mechanical compaction to concentrate matrix content and improve the compressive properties of such constructs. The inclusion of COL+PG improved the compressive properties of hydrogel constructs compared with PG or COL alone. Mechanical compaction increased the PG and COL concentrations in and compressive stiffness of the constructs. Chondrocytes included in the constructs maintained high viability after compaction. These results support the concepts that the assembly of cartilaginous constructs with COL+PG and application of mechanical compaction enhance the ECM content and compressive properties of engineered cartilaginous constructs.
Get full access to this article
View all access options for this article.
References
1.
BuckwalterJ.A., MankinH.J.Articular cartilage. Part I: tissue design and chondrocyte-matrix interactions. J Bone Joint Surg Am, 79-A:600. 1997.
2.
RoughleyP.J.The structure and function of cartilage proteoglycans. Eur Cell Mater, 12:92. 2006.
3.
MaroudasA.Balance between swelling pressure and collagen tension in normal and degenerate cartilage. Nature, 260:808. 1976.
4.
HanE.H., ChenS.S., KlischS.M., SahR.L.Contribution of proteoglycan osmotic swelling pressure to the compressive properties of articular cartilage. Biophys J, 101:916. 2011.
5.
BuckwalterJ.A., MankinH.J.Articular cartilage. Part II: degeneration and osteoarthrosis, repair, regeneration, and transplantation. J Bone Joint Surg Am, 79-A:612. 1997.
6.
WilliamsonA.K., ChenA.C., SahR.L.Compressive properties and function-composition relationships of developing bovine articular cartilage. J Orthop Res, 19:1113. 2001.
7.
TempleM.M., BaeW.C., ChenM.Q., LotzM., AmielD., CouttsR.D.et al.Age- and site-associated biomechanical weakening of human articular cartilage of the femoral condyle. Osteoarthritis Cartilage, 15:1042. 2007.
8.
ChenA.C., MasudaK., NakagawaK., WongV.W., PfisterB.E., ThonarE.J.et al.Tissue engineered cartilage from adult human chondrocytes: biomechanical properties and function-composition relationships. Trans Orthop Res Soc, 28:945. 2003.
9.
NgK.W., KuglerL.E., DotyS.B., AteshianG.A., HungC.T.Scaffold degradation elevates the collagen content and dynamic compressive modulus in engineered articular cartilage. Osteoarthritis Cartilage, 17:220. 2009.
10.
Vunjak-NovakovicG., MartinI., ObradovicB., TreppoS., GrodzinskyA.J., LangerR.et al.Bioreactor cultivation conditions modulate the composition and mechanical properties of tissue-engineered cartilage. J Orthop Res, 17:130. 1999.
11.
KleinT.J., MaldaJ., SahR.L., HutmacherD.W.Tissue engineering of articular cartilage with biomimetic zones. Tissue Eng Part B, 15:143. 2009.
12.
VinatierC., MrugalaD., JorgensenC., GuicheuxJ., NoelD.Cartilage engineering: a crucial combination of cells, biomaterials and biofactors. Trends Biotechnol, 27:307. 2009.
13.
HanE.H., WilenskyL.M., SchumacherB.L., ChenA.C., MasudaK., SahR.L.Tissue engineering by molecular disassembly and reassembly: biomimetic retention of mechanically functional aggrecan in hydrogel. Tissue Eng Part C Methods, 16:1471. 2010.
14.
AsanbaevaA., TamJ., SchumacherB.L., KlischS.M., MasudaK., SahR.L.Articular cartilage tensile integrity: modulation by matrix depletion is maturation-dependent. Arch Biochem Biophys, 474:175. 2008.
15.
NatoliR.M., ResponteD.J., LuB.Y., AthanasiouK.A.Effects of multiple chondroitinase ABC applications on tissue engineered articular cartilage. J Orthop Res, 27:949. 2009.
16.
BianL., CrivelloK.M., NgK.W., XuD., WilliamsD.Y., AteshianG.A.et al.Influence of temporary chondroitinase ABC-induced glycosaminoglycan suppression on maturation of tissue-engineered cartilage. Tissue Eng Part A, 15:2065. 2009.
17.
SajderaS.W., HascallV.C.Proteinpolysaccharide complex from bovine nasal cartilage. A comparison of low and high shear extraction procedures. J Biol Chem, 244:77. 1969.
18.
HeinegardD., SommarinY.Isolation and characterization of proteoglycans. Methods Enzymol, 144:305. 1987.
19.
TangL-H, RosenbergL., ReeinerA., PooleA.R.Proteoglycans from bovine nasal and articular cartilage: properties of a soluble form of link protein. J Biol Chem, 254:10523. 1979.
20.
SahR.L., GrodzinskyA.J., PlaasA.H., SandyJ.D.Effects of tissue compression on the hyaluronate-binding properties of newly synthesized proteoglycans in cartilage explants. Biochem J, 267:803. 1990.
21.
FrankE.H., GrodzinskyA.J., KoobT.J., EyreD.R.Streaming potentials: a sensitive index of enzymatic degradation in articular cartilage. J Orthop Res, 5:497. 1987.
22.
ChenA.C., NguyenT.T., SahR.L.Streaming potentials during the confined compression creep test of normal and proteoglycan-depleted cartilage. Ann Biomed Eng, 25:269. 1997.
23.
WoessnerJ.F.The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys, 93:440. 1961.
24.
FarndaleR.W., ButtleD.J., BarrettA.J.Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta, 883:173. 1986.
25.
ScottJ.E., DorlingJ.Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemie, 5:221. 1965.
26.
Hsieh-BonasseraN.D., WuI., LinJ.K., SchumacherB.L., ChenA.C., MasudaK.et al.Expansion and redifferentiation of chondrocytes from osteoarthritic cartilage: cells for human cartilage tissue engineering. Tissue Eng Part A, 15:3513. 2009.
27.
BasserP.J., SchneidermanR., BankR.A., WachtelE., MaroudasA.Mechanical properties of the collagen network in human articular cartilage as measured by osmotic stress technique. Arch Biochem Biophys, 351:207. 1998.
28.
BuschmannM.D., GrodzinskyA.J.A molecular model of proteoglycan-associated electrostatic forces in cartilage mechanics. J Biomech Eng, 117:179. 1995.
29.
MasudaK., SahR.L., HejnaM.J., ThonarE.J.-M.A.A novel two-step method for the formation of tissue-engineered cartilage by mature bovine chondrocytes: the alginate-recovered-chondrocyte (ARC) method. J Orthop Res, 21:139. 2003.
30.
KleinT.J., SchumacherB.L., BlewisM.E., SchmidtT.A., VoegtlineM.S., ThonarEJ-MAet al.Tailoring secretion of proteoglycan 4 (PRG4) in tissue-engineered cartilage. Tissue Eng, 12:1429. 2006.
31.
KatzE.P., WachtelE.J., MaroudasA.Extrafibrillar proteoglycans osmotically regulate the molecular packing of collagen in cartilage. Biochim Biophys Acta, 882:136. 1986.
KleinT.J., SchumacherB.L., SchmidtT.A., LiK.W., VoegtlineM.S., MasudaK.et al.Tissue engineering of stratified articular cartilage from chondrocyte subpopulations. Osteoarthritis Cartilage, 11:595. 2003.
34.
VeilleuxN.H., YannasI.V., SpectorM.Effect of passage number and collagen type on the proliferative, biosynthetic, and contractile activity of adult canine articular chondrocytes in type I and II collagen-glycosaminoglycan matrices in vitro. Tissue Eng, 10:119. 2004.
35.
GaloisL., HutasseS., CortialD., RousseauC.F., GrossinL., RonziereM.C.et al.Bovine chondrocyte behaviour in three-dimensional type I collagen gel in terms of gel contraction, proliferation and gene expression. Biomaterials, 27:79. 2006.
36.
LeeC.R., BreinanH.A., NehrerS., SpectorM.Articular cartilage chondrocytes in type I and type II collagen-GAG matrices exhibit contractile behavior in vitro. Tissue Eng, 6:555. 2000.
37.
ElderB.D., EleswarapuS.V., AthanasiouK.A.Extraction techniques for the decellularization of tissue engineered articular cartilage constructs. Biomaterials, 30:3749. 2009.
38.
NicodemusG.D., BryantS.J.Cell encapsulation in biodegradable hydrogels for tissue engineering applications. Tissue Eng Part B Rev, 14:149. 2008.
39.
KohJ.L., WirsingK., LautenschlagerE., ZhangL.O.The effect of graft height mismatch on contact pressure following osteochondral grafting. A biomechanical study. Am J Sports Med, 32:317. 2004.
JadinK.D., BaeW.C., SchumacherB.L., SahR.L.Three-dimensional (3-D) imaging of chondrocytes in articular cartilage: growth-associated changes in cell organization. Biomaterials, 28:230. 2007.
47.
MankinH.J.Localization of tritiated thymidine in articular cartilage of rabbits. I. growth in immature cartilage. J Bone Joint Surg Am, 44-A:682. 1962.
48.
AkizukiS., MowV.C., MullerF., PitaJ.C., HowellD.S., ManicourtD.H.Tensile properties of human knee joint cartilage: I. influence of ionic conditions, weight bearing, and fibrillation on the tensile modulus. J Orthop Res, 4:379. 1986.
AignerT., StoveJ.Collagens-major component of the physiological cartilage matrix, major target of cartilage degeneration, major tool in cartilage repair. Adv Drug Deliv Rev, 55:1569. 2003.
51.
VargheseS., HwangN.S., CanverA.C., TheprungsirikulP., LinD.W., ElisseeffJ.Chondroitin sulfate based niches for chondrogenic differentiation of mesenchymal stem cells. Matrix Biol, 27:12. 2008.
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.
