Matrix forming characteristics of inner and outer human meniscus cells on 3D collagen scaffolds under normal and low oxygen tensions

BMC Musculoskeletal Disorders

Adetola Adesida¹  Hasan Uludag¹  Nadr Jomha¹  Roger Croutze¹ 

[1]Department of Surgery, Division of Orthopaedic Surgery, Laboratory of Stem Cell Biology and Orthopaedic Tissue Engineering, University of Alberta, Faculty of Medicine and Dentistry, 3.002E Li Ka Shing Centre for Health Research Innovation, Edmonton, AB T6G 2E1, Canada

关键词: Tissue engineering;    Scaffold;    Collagen;    Hypoxia;    Normoxia;    Oxygen tension;    Meniscus fibrochondrocyte;    Meniscus;   

Others  :  1129045
DOI  :  10.1186/1471-2474-14-353

 received in 2013-06-28, accepted in 2013-11-29,  发布年份 2013

【 摘 要 】

Background

Limited intrinsic healing potential of the meniscus and a strong correlation between meniscal injury and osteoarthritis have prompted investigation of surgical repair options, including the implantation of functional bioengineered constructs. Cell-based constructs appear promising, however the generation of meniscal constructs is complicated by the presence of diverse cell populations within this heterogeneous tissue and gaps in the information concerning their response to manipulation of oxygen tension during cell culture.

Methods

Four human lateral menisci were harvested from patients undergoing total knee replacement. Inner and outer meniscal fibrochondrocytes (MFCs) were expanded to passage 3 in growth medium supplemented with basic fibroblast growth factor (FGF-2), then embedded in porous collagen type I scaffolds and chondrogenically stimulated with transforming growth factor β3 (TGF-β3) under 21% (normal or normoxic) or 3% (hypoxic) oxygen tension for 21 days. Following scaffold culture, constructs were analyzed biochemically for glycosaminoglycan production, histologically for deposition of extracellular matrix (ECM), as well as at the molecular level for expression of characteristic mRNA transcripts.

Results

Constructs cultured under normal oxygen tension expressed higher levels of collagen type II (p = 0.05), aggrecan (p < 0.05) and cartilage oligomeric matrix protein, (COMP) (p < 0.05) compared to hypoxic expanded and cultured constructs. Accumulation of ECM rich in collagen type II and sulfated proteoglycan was evident in normoxic cultured scaffolds compared to those under low oxygen tension. There was no significant difference in expression of these genes between scaffolds seeded with MFCs isolated from inner or outer regions of the tissue following 21 days chondrogenic stimulation (p > 0.05).

Conclusions

Cells isolated from inner and outer regions of the human meniscus demonstrated equivalent differentiation potential toward chondrogenic phenotype and ECM production. Oxygen tension played a key role in modulating the redifferentiation of meniscal fibrochondrocytes on a 3D collagen scaffold in vitro.

【 授权许可】

   
2013 Croutze et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Aagaard H, Verdonk R: Function of the normal meniscus and consequences of meniscal resection. Scand J Med Sci Sports 1999, 9(3):134-140.
• [2]McDermott ID, Amis AA: The consequences of meniscectomy. J Bone Joint Surg Br 2006, 88(12):1549-1556.
• [3]Sanchez-Adams J: The knee meniscus: a complex tissue of diverse cells. Cell Mol Bioeng 2009, 2:332-340.
• [4]Chevrier A, Nelea M, Hurtig MB, Hoemann CD, Buschmann MD: Meniscus structure in human, sheep, and rabbit for animal models of meniscus repair. J Orthop Res 2009, 27(9):1197-1203.
• [5]Nakata K, Shino K, Hamada M, Mae T, Miyama T, Shinjo H, Horibe S, Tada K, Ochi T, Yoshikawa H: Human meniscus cell: characterization of the primary culture and use for tissue engineering. Clin Orthop Relat Res 2001, 391:S208-218.
• [6]Fukazawa I, Hatta T, Uchio Y, Otani H: Development of the meniscus of the knee joint in human fetuses. Congenit Anom (Kyoto) 2009, 49(1):27-32.
• [7]Arnoczky SP, Warren RF: Microvasculature of the human meniscus. Am J Sports Med 1982, 10(2):90-95.
• [8]Arnoczky SP, Warren RF: The microvasculature of the meniscus and its response to injury. An experimental study in the dog. Am J Sports Med 1983, 11(3):131-141.
• [9]King D: The healing of semilunar cartilage. J Bone Joint Surg 1936, 18B:333-342.
• [10]Gu YL, Wang YB: Treatment of meniscal injury: a current concept review. Chin J Traumatol 2010, 13(6):370-376.
• [11]Clayton RA, Court-Brown CM: The epidemiology of musculoskeletal tendinous and ligamentous injuries. Injury 2008, 39(12):1338-1344.
• [12]Berthiaume MJ, Raynauld JP, Martel-Pelletier J, Labonte F, Beaudoin G, Bloch DA, Choquette D, Haraoui B, Altman RD, Hochberg M, et al.: Meniscal tear and extrusion are strongly associated with progression of symptomatic knee osteoarthritis as assessed by quantitative magnetic resonance imaging. Ann Rheum Dis 2005, 64(4):556-563.
• [13]Roos H, Lauren M, Adalberth T, Roos EM, Jonsson K, Lohmander LS: Knee osteoarthritis after meniscectomy: prevalence of radiographic changes after twenty-one years, compared with matched controls. Arthritis Rheum 1998, 41(4):687-693.
• [14]McNicholas MJ, Pengas IP, Assiotis A, Nash W, Hatcher J, Banks J: Total meniscectomy in adolescents: a 40-year follow-up. J Bone Joint Surg Br 2012, 94(12):1649-1654.
• [15]Fairbank TJ: Knee joint changes after meniscectomy. J Bone Joint Surg Br 1948, 30B(4):664-670.
• [16]Lee AS, Kang RW, Kroin E, Verma NN, Cole BJ: Allograft meniscus transplantation. Sports Med Arthrosc 2012, 20(2):106-114.
• [17]Zhang Z, Arnold JA, Williams T, McCann B: Repairs by trephination and suturing of longitudinal injuries in the avascular area of the meniscus in goats. Am J Sports Med 1995, 23(1):35-41.
• [18]Longo UG, Campi S, Romeo G, Spiezia F, Maffulli N, Denaro V: Biological strategies to enhance healing of the avascular area of the meniscus. Stem Cells Int 2012, 2012:528359.
• [19]Arnoczky SP, Warren RF, Spivak JM: Meniscal repair using an exogenous fibrin clot. An experimental study in dogs. J Bone Joint Surg Am 1988, 70(8):1209-1217.
• [20]Kon E, Filardo G, Tschon M, Fini M, Giavaresi G, Marchesini Reggiani L, Chiari C, Nehrer S, Martin I, Salter DM, et al.: Tissue engineering for total meniscal substitution: animal study in sheep model–results at 12 months. Tissue Eng Part A 2012, 18(15–16):1573-1582.
• [21]Moriguchi Y, Tateishi K, Ando W, Shimomura K, Yonetani Y, Tanaka Y, Kita K, Hart DA, Gobbi A, Shino K, et al.: Repair of meniscal lesions using a scaffold-free tissue-engineered construct derived from allogenic synovial MSCs in a miniature swine model. Biomaterials 2013, 34(9):2185-2193.
• [22]Khoury MA, Goldberg VM, Stevenson S: Demonstration of HLA and ABH antigens in fresh and frozen human menisci by immunohistochemistry. J Orthop Res 1994, 12(6):751-757.
• [23]Verdonk P, Beaufils P, Bellemans J, Djian P, Heinrichs EL, Huysse W, Laprell H, Siebold R, Verdonk R, the Actifit Study G: Successful treatment of painful irreparable partial meniscal defects with a polyurethane scaffold: two-year safety and clinical outcomes. Am J Sports Med 2012, 40(4):844-853.
• [24]Gunja NJ, Athanasiou KA: Passage and reversal effects on gene expression of bovine meniscal fibrochondrocytes. Arthritis Res Ther 2007, 9(5):R93. BioMed Central Full Text
• [25]Adesida AB, Grady LM, Khan WS, Hardingham TE: The matrix-forming phenotype of cultured human meniscus cells is enhanced after culture with fibroblast growth factor 2 and is further stimulated by hypoxia. Arthritis Res Ther 2006, 8(3):R61. BioMed Central Full Text
• [26]Pereira H, Frias AM, Oliveira JM, Espregueira-Mendes J, Reis RL: Tissue engineering and regenerative medicine strategies in meniscus lesions. Arthroscopy 2011, 27(12):1706-1719.
• [27]Mueller SM, Shortkroff S, Schneider TO, Breinan HA, Yannas IV, Spector M: Meniscus cells seeded in type I and type II collagen-GAG matrices in vitro. Biomaterials 1999, 20(8):701-709.
• [28]Gruber HE, Mauerhan D, Chow Y, Ingram JA, Norton HJ, Hanley EN Jr, Sun Y: Three-dimensional culture of human meniscal cells: extracellular matrix and proteoglycan production. BMC Biotechnol 2008, 8:54. BioMed Central Full Text
• [29]Adesida AB, Mulet-Sierra A, Laouar L, Jomha NM: Oxygen tension is a determinant of the matrix-forming phenotype of cultured human meniscal fibrochondrocytes. PLoS One 2012, 7(6):e39339.
• [30]Malda J, Martens DE, Tramper J, van Blitterswijk CA, Riesle J: Cartilage tissue engineering: controversy in the effect of oxygen. Crit Rev Biotechnol 2003, 23(3):175-194.
• [31]Farndale RW, Buttle DJ, Barrett AJ: Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta 1986, 883(2):173-177.
• [32]Adesida AB, Grady LM, Khan WS, Millward-Sadler SJ, Salter DM, Hardingham TE: Human meniscus cells express hypoxia inducible factor-1alpha and increased SOX9 in response to low oxygen tension in cell aggregate culture. Arthritis Res Ther 2007, 9(4):R69. BioMed Central Full Text
• [33]Khan WS, Adesida AB, Hardingham TE: Hypoxic conditions increase hypoxia-inducible transcription factor 2alpha and enhance chondrogenesis in stem cells from the infrapatellar fat pad of osteoarthritis patients. Arthritis Res Ther 2007, 9(3):R55. BioMed Central Full Text
• [34]Murdoch AD, Grady LM, Ablett MP, Katopodi T, Meadows RS, Hardingham TE: Chondrogenic differentiation of human bone marrow stem cells in transwell cultures: generation of scaffold-free cartilage. Stem Cells 2007, 25(11):2786-2796.
• [35]Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001, 25(4):402-408.
• [36]Mauck RL, Martinez-Diaz GJ, Yuan X, Tuan RS: Regional multilineage differentiation potential of meniscal fibrochondrocytes: implications for meniscus repair. Anat Rec (Hoboken) 2007, 290(1):48-58.
• [37]Martin I, Vunjak-Novakovic G, Yang J, Langer R, Freed LE: Mammalian chondrocytes expanded in the presence of fibroblast growth factor 2 maintain the ability to differentiate and regenerate three-dimensional cartilaginous tissue. Exp Cell Res 1999, 253(2):681-688.
• [38]Barbero A, Ploegert S, Heberer M, Martin I: Plasticity of clonal populations of dedifferentiated adult human articular chondrocytes. Arthritis Rheum 2003, 48(5):1315-1325.
• [39]Sekiya I, Koopman P, Tsuji K, Mertin S, Harley V, Yamada Y, Shinomiya K, Nifuji A, Noda M: Dexamethasone enhances SOX9 expression in chondrocytes. J Endocrinol 2001, 169(3):573-579.
• [40]de Crombrugghe B, Lefebvre V, Behringer RR, Bi W, Murakami S, Huang W: Transcriptional mechanisms of chondrocyte differentiation. Matrix Biol 2000, 19(5):389-394.
• [41]Sekiya I, Tsuji K, Koopman P, Watanabe H, Yamada Y, Shinomiya K, Nifuji A, Noda M: SOX9 enhances aggrecan gene promoter/enhancer activity and is up-regulated by retinoic acid in a cartilage-derived cell line, TC6. J Biol Chem 2000, 275(15):10738-10744.
• [42]Aigner T, Gebhard PM, Schmid E, Bau B, Harley V, Poschl E: SOX9 expression does not correlate with type II collagen expression in adult articular chondrocytes. Matrix Biol 2003, 22(4):363-372.
• [43]Duval E, Leclercq S, Elissalde JM, Demoor M, Galera P, Boumediene K: Hypoxia-inducible factor 1alpha inhibits the fibroblast-like markers type I and type III collagen during hypoxia-induced chondrocyte redifferentiation: hypoxia not only induces type II collagen and aggrecan, but it also inhibits type I and type III collagen in the hypoxia-inducible factor 1alpha-dependent redifferentiation of chondrocytes. Arthritis Rheum 2009, 60(10):3038-3048.
• [44]Lafont JE, Talma S, Murphy CL: Hypoxia-inducible factor 2alpha is essential for hypoxic induction of the human articular chondrocyte phenotype. Arthritis Rheum 2007, 56(10):3297-3306.