Research Article

Departments of Orthopedic Surgery & Biomedical Engineering, Duke University Medical Center, Durham, NC 27710, USA

Institute of Biomedical Engineering, College of Medicine & College of Engineering, National Taiwan University, Taipei, 100 Taiwan

Department of Surgery, National Taiwan University Hospital & College of Medicine, Taipei, 100 Taiwan

Search for more papers by this author

Bradley T Estes

Departments of Orthopedic Surgery & Biomedical Engineering, Duke University Medical Center, Durham, NC 27710, USA

Search for more papers by this author

Tai-Horng Young

Institute of Biomedical Engineering, College of Medicine & College of Engineering, National Taiwan University, Taipei, 100 Taiwan

Search for more papers by this author

Farshid Guilak

† Author for correspondence

Search for more papers by this author

Email the corresponding author at guilak@duke.edu

Published Online:22 Dec 2010https://doi.org/10.2217/rme.10.87

View article

Abstract

Aim: To investigate the cell growth, matrix accumulation and mechanical properties of neocartilage formed by human or porcine articular chondrocytes on a porous, porcine cartilage-derived matrix (CDM) for use in cartilage tissue engineering. Materials & methods: We examined the physical properties, cell infiltration and matrix accumulation in different formulations of CDM and selected a CDM made of homogenized cartilage slurry as an appropriate scaffold for long-term culture of human and porcine articular chondrocytes. Results: The CDM scaffold supported growth and proliferation of both human and porcine chondrocytes. Histology and immunohistochemistry showed abundant cartilage-specific macromolecule deposition at day 28. Human chondrocytes migrated throughout the CDM, showing a relatively homogeneous distribution of new tissue accumulation, whereas porcine chondrocytes tended to form a proteoglycan-rich layer primarily on the surfaces of the scaffold. Human chondrocyte-seeded scaffolds had a significantly lower aggregate modulus and hydraulic permeability at day 28. Conclusions: These data show that a scaffold derived from native porcine articular cartilage can support neocartilage formation in the absence of exogenous growth factors. The overall characteristics and properties of the constructs depend on factors such as the concentration of CDM used, the porosity of the scaffold, and the species of chondrocytes.

Keywords:

Bibliography

1 Jackson DW, Simon TM, Aberman HM: Symptomatic articular cartilage degeneration: the impact in the new millennium. Clin. Orthop. Relat. Res. (391 Suppl.),S14–S25 (2001).
MedlineGoogle Scholar
2 Lynn AK, Brooks RA, Bonfield W, Rushton N: Repair of defects in articular joints. Prospects for material-based solutions in tissue engineering. J. Bone Joint Surg. Br.86(8),1093–1099 (2004).
Medline CASGoogle Scholar
3 Grande DA, Pitman MI, Peterson L, Menche D, Klein M: The repair of experimentally produced defects in rabbit articular cartilage by autologous chondrocyte transplantation. J. Orthop. Res.7(2),208–218 (1989).
Medline CASGoogle Scholar
4 Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L: Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N. Engl. J. Med.331(14),889–895 (1994).
Medline CASGoogle Scholar
5 Knutsen G, Engebretsen L, Ludvigsen TC et al.: Autologous chondrocyte implantation compared with microfracture in the knee. A randomized trial. J. Bone Joint Surg. Am.86-A(3),455–464 (2004).
MedlineGoogle Scholar
6 Awad HA, Halvorsen YD, Gimble JM, Guilak F: Effects of transforming growth factor-β1 and dexamethasone on the growth and chondrogenic differentiation of adipose-derived stromal cells. Tissue Eng.9(6),1301–1312 (2003).
Medline CASGoogle Scholar
7 Guilak F, Butler DL, Goldstein SA: Functional tissue engineering: the role of biomechanics in articular cartilage repair. Clin. Orthop. Relat. Res. (391 Suppl.),S295–S305 (2001).
MedlineGoogle Scholar
8 Aigner J, Tegeler J, Hutzler P et al.: Cartilage tissue engineering with novel nonwoven structured biomaterial based on hyaluronic acid benzyl ester. J. Biomed. Mater. Res.42(2),172–181 (1998).
Medline CASGoogle Scholar
9 Mierisch CM, Cohen SB, Jordan LC, Robertson PG, Balian G, Diduch DR: Transforming growth factor-β in calcium alginate beads for the treatment of articular cartilage defects in the rabbit. Arthroscopy18(8),892–900 (2002).
MedlineGoogle Scholar
10 Iwasaki N, Yamane ST, Majima T et al.: Feasibility of polysaccharide hybrid materials for scaffolds in cartilage tissue engineering: evaluation of chondrocyte adhesion to polyion complex fibers prepared from alginate and chitosan. Biomacromolecules5(3),828–833 (2004).
Medline CASGoogle Scholar
11 Domm C, Schunke M, Steinhagen J, Freitag S, Kurz B: Influence of various alginate brands on the redifferentiation of dedifferentiated bovine articular chondrocytes in alginate bead culture under high and low oxygen tension. Tissue Eng.10(11–12),1796–1805 (2004).
Medline CASGoogle Scholar
12 Buschmann MD, Gluzband YA, Grodzinsky AJ, Hunziker EB: Mechanical compression modulates matrix biosynthesis in chondrocyte/agarose culture. J. Cell Sci.108(Pt 4),1497–1508 (1995).
Medline CASGoogle Scholar
13 Mauck RL, Nicoll SB, Seyhan SL, Ateshian GA, Hung CT: Synergistic action of growth factors and dynamic loading for articular cartilage tissue engineering. Tissue Eng.9(4),597–611 (2003).
Medline CASGoogle Scholar
14 Mauck RL, Soltz MA, Wang CC et al.: Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. J. Biomech. Eng.122(3),252–260 (2000).
Medline CASGoogle Scholar
15 Nettles DL, Elder SH, Gilbert JA: Potential use of chitosan as a cell scaffold material for cartilage tissue engineering. Tissue Eng.8(6),1009–1016 (2002).
Medline CASGoogle Scholar
16 Cao Y, Rodriguez A, Vacanti M, Ibarra C, Arevalo C, Vacanti CA: Comparative study of the use of poly(glycolic acid), calcium alginate and pluronics in the engineering of autologous porcine cartilage. J. Biomater. Sci. Polym. Ed.9(5),475–487 (1998).
Medline CASGoogle Scholar
17 Chang SC, Rowley JA, Tobias G et al.: Injection molding of chondrocyte/alginate constructs in the shape of facial implants. J. Biomed. Mater. Res.55(4),503–511 (2001).
Medline CASGoogle Scholar
18 Fragonas E, Valente M, Pozzi-Mucelli M et al.: Articular cartilage repair in rabbits by using suspensions of allogenic chondrocytes in alginate. Biomaterials21(8),795–801 (2000).
Medline CASGoogle Scholar
19 Hendrickson DA, Nixon AJ, Grande DA et al.: Chondrocyte–fibrin matrix transplants for resurfacing extensive articular cartilage defects. J. Orthop. Res.12(4),485–497 (1994).
Medline CASGoogle Scholar
20 Kurz B, Domm C, Jin M, Sellckau R, Schunke M: Tissue engineering of articular cartilage under the influence of collagen I/III membranes and low oxygen tension. Tissue Eng.10(7–8),1277–1286 (2004).
Medline CASGoogle Scholar
21 Dausse Y, Grossin L, Miralles G et al.: Cartilage repair using new polysaccharidic biomaterials: macroscopic, histological and biochemical approaches in a rat model of cartilage defect. Osteoarthr. Cartil.11(1),16–28 (2003).
Medline CASGoogle Scholar
22 Kang Y, Yang J, Khan S, Anissian L, Ameer GA: A new biodegradable polyester elastomer for cartilage tissue engineering. J. Biomed. Mater. Res. A77(2),331–339 (2006).
MedlineGoogle Scholar
23 Bryant SJ, Anseth KS: Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels. J. Biomed. Mater. Res.59(1),63–72 (2002).
Medline CASGoogle Scholar
24 Lee WK, Ichi T, Ooya T, Yamamoto T, Katoh M, Yui N: Novel poly(ethylene glycol) scaffolds crosslinked by hydrolyzable polyrotaxane for cartilage tissue engineering. J. Biomed. Mater. Res. A67(4),1087–1092 (2003).
MedlineGoogle Scholar
25 Freed LE, Vunjak-Novakovic G, Langer R: Cultivation of cell-polymer cartilage implants in bioreactors. J. Cell Biochem.51(3),257–264 (1993).
Medline CASGoogle Scholar
26 Martin I, Obradovic B, Treppo S et al.: Modulation of the mechanical properties of tissue engineered cartilage. Biorheology37(1–2),141–147 (2000).
Medline CASGoogle Scholar
27 Urist MR: Bone: formation by autoinduction. Science150(698),893–899 (1965).
Medline CASGoogle Scholar
28 Badylak SF: The extracellular matrix as a biologic scaffold material. Biomaterials28(25),3587–3593 (2007).
Medline CASGoogle Scholar
29 Badylak SF, Freytes DO, Gilbert TW: Extracellular matrix as a biological scaffold material: structure and function. Acta Biomater.5(1),1–13 (2009).
Medline CASGoogle Scholar
30 Chang Y, Lee MH, Liang HC, Hsu CK, Sung HW: Acellular bovine pericardia with distinct porous structures fixed with genipin as an extracellular matrix. Tissue Eng.10(5–6),881–892 (2004).
Medline CASGoogle Scholar
31 Ott HC, Matthiesen TS, Goh SK et al.: Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat. Med.14(2),213–221 (2008).
Medline CASGoogle Scholar
32 Little D, Guilak F, Ruch DS: Ligament-derived matrix stimulates a ligamentous phenotype in human adipose-derived stem cells. Tissue Eng. Part A16(7),2307–2319 (2010).
Medline CASGoogle Scholar
33 Ponce Marquez S, Martinez VS, McIntosh Ambrose W et al.: Decellularization of bovine corneas for tissue engineering applications. Acta Biomater.5(6),1839–1847 (2009).
MedlineGoogle Scholar
34 Hwang NS, Varghese S, Puleo C, Zhang Z, Elisseeff J: Morphogenetic signals from chondrocytes promote chondrogenic and osteogenic differentiation of mesenchymal stem cells. J. Cell Physiol.212(2),281–284 (2007).
Medline CASGoogle Scholar
35 Cheng NC, Estes BT, Awad HA, Guilak F: Chondrogenic differentiation of adipose-derived adult stem cells by a porous scaffold derived from native articular cartilage extracellular matrix. Tissue Eng. Part A15(2),231–241 (2009).
Medline CASGoogle Scholar
36 Diekman BO, Rowland CR, Caplan AI, Lennon D, Guilak F: Chondrogenesis of adult stem cells from adipose tissue and bone marrow: induction by growth factors and cartilage derived matrix. Tissue Eng. Part A16(2),523–533 (2009).
Google Scholar
37 Lin YC, Tan FJ, Marra KG, Jan SS, Liu DC: Synthesis and characterization of collagen/hyaluronan/chitosan composite sponges for potential biomedical applications. Acta Biomater.5(7),2591–2600 (2009).
Medline CASGoogle Scholar
38 Guilak F, Erickson GR, Ting-Beall HP: The effects of osmotic stress on the viscoelastic and physical properties of articular chondrocytes. Biophys. J.82(2),720–727 (2002).
Medline CASGoogle Scholar
39 Darling EM, Athanasiou KA: Rapid phenotypic changes in passaged articular chondrocyte subpopulations. J. Orthop. Res.23(2),425–432 (2005).
Medline CASGoogle Scholar
40 Farndale RW, Buttle DJ, Barrett AJ: Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim. Biophys. Acta883(2),173–177 (1986).
Medline CASGoogle Scholar
41 Woessner JF Jr: The determination of hydroxyproline in tissue and protein samples containing small proportions of this amino acid. Arch. Biochem. Biophys.93,440–447 (1961).
Medline CASGoogle Scholar
42 Mow VC, Kuei SC, Lai WM, Armstrong CG: Biphasic creep and stress-relaxation of articular-cartilage in compression – theory and experiments. J. Biomech. Eng.102(1),73–84 (1980).
Medline CASGoogle Scholar
43 Bachrach NM, Mow VC, Guilak F: Incompressibility of the solid matrix of articular cartilage under high hydrostatic pressures. J. Biomech.31(5),445–451 (1998).
Medline CASGoogle Scholar
44 Bi X, Yang X, Bostrom MP, Camacho NP: Fourier transform infrared imaging spectroscopy investigations in the pathogenesis and repair of cartilage. Biochim. Biophys. Acta1758(7),934–941 (2006).
Medline CASGoogle Scholar
45 Potter K, Kidder LH, Levin IW, Lewis EN, Spencer RG: Imaging of collagen and proteoglycan in cartilage sections using Fourier transform infrared spectral imaging. Arthritis Rheum.44(4),846–855 (2001).
Medline CASGoogle Scholar
46 Basad E, Ishaque B, Bachmann G, Sturz H, Steinmeyer J: Matrix-induced autologous chondrocyte implantation versus microfracture in the treatment of cartilage defects of the knee: a 2-year randomised study. Knee Surg. Sports Traumatol. Arthrosc.18(4),519–527 (2010).
MedlineGoogle Scholar
47 Kuo CK, Li WJ, Mauck RL, Tuan RS: Cartilage tissue engineering: its potential and uses. Curr. Opin. Rheumatol.18(1),64–73 (2006).
MedlineGoogle Scholar
48 Wang Y, Kim UJ, Blasioli DJ, Kim HJ, Kaplan DL: In vitro cartilage tissue engineering with 3D porous aqueous-derived silk scaffolds and mesenchymal stem cells. Biomaterials26(34),7082–7094 (2005).
Medline CASGoogle Scholar
49 Leddy HA, Awad HA, Guilak F: Molecular diffusion in tissue-engineered cartilage constructs: effects of scaffold material, time, and culture conditions. J. Biomed. Mater. Res. B Appl. Biomater.70(2),397–406 (2004).
MedlineGoogle Scholar
50 Macchiarini P, Jungebluth P, Go T et al.: Clinical transplantation of a tissue-engineered airway. Lancet372(9655),2023–2030 (2008).
MedlineGoogle Scholar
51 Yang Q, Peng J, Guo Q et al.: A cartilage ECM-derived 3-D porous acellular matrix scaffold for in vivo cartilage tissue engineering with PKH26-labeled chondrogenic bone marrow-derived mesenchymal stem cells. Biomaterials29(15),2378–2387 (2008).
Medline CASGoogle Scholar
52 Bobick BE, Chen FH, Le AM, Tuan RS: Regulation of the chondrogenic phenotype in culture. Birth Defects Res. C Embryo Today87(4),351–371 (2009).
Medline CASGoogle Scholar
53 Vincent TL, McLean CJ, Full LE, Peston D, Saklatvala J: FGF-2 is bound to perlecan in the pericellular matrix of articular cartilage, where it acts as a chondrocyte mechanotransducer. Osteoarthr. Cartil.15(7),752–763 (2007).
Medline CASGoogle Scholar
54 Zhu Y, Oganesian A, Keene DR, Sandell LJ: Type IIA procollagen containing the cysteine-rich amino propeptide is deposited in the extracellular matrix of prechondrogenic tissue and binds to TGF-β1 and BMP-2. J. Cell Biol.144(5),1069–1080 (1999).
Medline CASGoogle Scholar
55 Betre H, Ong SR, Guilak F, Chilkoti A, Fermor B, Setton LA: Chondrocytic differentiation of human adipose-derived adult stem cells in elastin-like polypeptide. Biomaterials27(1),91–99 (2006).
Medline CASGoogle Scholar
56 Boskey A, Pleshko Camacho N: FT-IR imaging of native and tissue-engineered bone and cartilage. Biomaterials28(15),2465–2478 (2007).
Medline CASGoogle Scholar
57 O’Brien FJ, Harley BA, Yannas IV, Gibson LJ: The effect of pore size on cell adhesion in collagen–GAG scaffolds. Biomaterials26(4),433–441 (2005).
MedlineGoogle Scholar
58 O’Brien FJ, Harley BA, Waller MA, Yannas IV, Gibson LJ, Prendergast PJ: The effect of pore size on permeability and cell attachment in collagen scaffolds for tissue engineering. Technol. Health Care15(1),3–17 (2007).
MedlineGoogle Scholar
59 Schulze-Tanzil G, Muller RD, Kohl B et al.: Differing in vitro biology of equine, ovine, porcine and human articular chondrocytes derived from the knee joint: an immunomorphological study. Histochem. Cell Biol.131(2),219–229 (2009).
Medline CASGoogle Scholar
60 Akens MK, Hurtig MB: Influence of species and anatomical location on chondrocyte expansion. BMC Musculoskelet. Disord.6,23 (2005).
MedlineGoogle Scholar
61 Giannoni P, Crovace A, Malpeli M et al.: Species variability in the differentiation potential of in vitro-expanded articular chondrocytes restricts predictive studies on cartilage repair using animal models. Tissue Eng.11(1–2),237–248 (2005).
Medline CASGoogle Scholar
62 Diekman BO, Rowland CR, Caplan AI, Lennon D, Guilak F: Chondrogenesis of adult stem cells from adipose tissue and bone marrow: induction by growth factors and cartilage derived matrix. Tissue Eng. Part A16(2),523–533 (2009).
Google Scholar
63 Valonen PK, Moutos FT, Kusanagi A et al.: In vitro generation of mechanically functional cartilage grafts based on adult human stem cells and 3D-woven poly(ε-caprolactone) scaffolds. Biomaterials31(8),2193–2200 (2010).
Medline CASGoogle Scholar
64 Athanasiou KA, Rosenwasser MP, Buckwalter JA, Malinin TI, Mow VC: Interspecies comparisons of in situ intrinsic mechanical properties of distal femoral cartilage. J. Orthop. Res.9(3),330–340 (1991).
Medline CASGoogle Scholar
65 Cavallo C, Desando G, Facchini A, Grigolo B: Chondrocytes from patients with osteoarthritis express typical extracellular matrix molecules once grown onto a three-dimensional hyaluronan-based scaffold. J. Biomed. Mater. Res. A93(1),86–95 (2009).
Google Scholar
66 Seda Tigli R, Ghosh S, Laha MM et al.: Comparative chondrogenesis of human cell sources in 3D scaffolds. J. Tissue Eng. Regen. Med.3(5),348–360 (2009).
MedlineGoogle Scholar
67 Freed LE, Engelmayr J, Borenstein JT, Moutos FT, Guilak F: Advanced material strategies for tissue engineering scaffolds. Adv. Mater.21(32–33),3410–3418 (2009).
Medline CASGoogle Scholar
68 Discher DE, Mooney DJ, Zandstra PW: Growth factors, matrices, and forces combine and control stem cells. Science324(5935),1673–1677 (2009).
Medline CASGoogle Scholar
69 Freed LE, Guilak F, Guo XE et al.: Advanced tools for tissue engineering: scaffolds, bioreactors, and signaling. Tissue Eng.12(12),3285–3305 (2006).
Medline CASGoogle Scholar
70 Guilak F, Awad HA, Fermor B, Leddy HA, Gimble JM: Adipose-derived adult stem cells for cartilage tissue engineering. Biorheology41(3–4),389–399 (2004).
Medline CASGoogle Scholar
71 Estes BT, Gimble JM, Guilak F: Mechanical signals as regulators of stem cell fate. Curr. Top Dev. Biol.60,91–126 (2004).
Medline CASGoogle Scholar

Vol. 6, No. 1

Metrics

Downloaded 262 times

History

Published online 22 December 2010

Published in print January 2011

Information

Keywords

Acknowledgements

The authors acknowledge the technical assistance from the staff of the Eighth Core Lab of National Taiwan University Hospital.

Financial & competing interests disclosure

The project was partially supported by Osteotech Inc., NIH grants AR48852, AR48182, AR055042, AG15768 and AR50245, grants from the National Science Council, Taiwan (NSC 97–2320-B-002–010) and National Taiwan University Hospital (98-N1198). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

PDF download

Engineered cartilage using primary chondrocytes cultured in a porous cartilage-derived matrix

Abstract

Bibliography