CellBio  Vol.2 No.4 , December 2013
The Knee Joint Tissues Differ Significantly in TGFβ1 Expression and Its Sensitivity
Abstract: The knee joint is the largest and most complex joint in the human body. In this study, we investigated TGFβ1 expression in the outer meniscus, inner meniscus and articular cartilage of rabbit and human knee tissue (outer and inner menisci) in order to determine the potential role of this factor in normal meniscal function. We also examined the potential of TGF-β1 stimulation to promote tissue regeneration in the two different regions of rabbit knee meniscus tissue. Immunohistochemical investigations of TGF-β1 were performed on rabbit and human knee tissue. The rabbit outer, inner and articular cartilage cells were culture and stimulated with TGF-β1 followed by cell proliferation assay and extracellular matrix analysis. Regulatory studies were performed using TGF-β1 inhibitors SB-431542 and PD98059. Gene expression was analyzed by quantitative polymerase chain reaction. We found marked regional variation in the expression of TGF-β1 in rabbit and human knee. TGF-β1 expressions are relatively greater in the outer meniscus than inner meniscus. Furthermore, we found that exogenous TGF-β1 stimulation increased cell proliferation and aggrecan synthesis more so in the outer than in the inner meniscus. Articular cartilage tissue shows moderate levels of cell proliferation and ECM synthesis when compared with outer and inner meniscus. These findings suggest that growth factors used to enhance the repair and regeneration of meniscal tissue should be tailored to enhance region-specific variation in cell proliferation and extracellular matrix synthesis.
Cite this paper: Fulzele, S. , Hunter, M. , Sangani, R. , Chutkan, N. , Isales, C. and W. Hamrick, M. (2013) The Knee Joint Tissues Differ Significantly in TGFβ1 Expression and Its Sensitivity. CellBio, 2, 192-199. doi: 10.4236/cellbio.2013.24022.

[1]   P. Ghosh, Y. Numata, S. Smith, R. Read, S. Armstrong and K. Johnson, “The Metabolic Response of Articular Cartilage to Abnormal Mechanical Loading Induced by Medial or Lateral Meniscectomy,” Agents and Actions Supplement, Vol. 39, 1993, pp. 89-93.

[2]   J. Sanchez-Adams and K. Athanasiou, “The Knee Meniscus: A Complex Tissue of Diverse Cells,” Cellular and Molecular Bioengineering, Vol. 2, No. 3, 2009, pp. 332-340.

[3]   S. P. Arnoczky, “Building a Meniscus. Biologic Considerations,” Clinical Orthopaedics and Related Research, No. 367, 1999, pp. S244-S253.

[4]   M. Cucchiarini, S. Schetting, E. F. Terwilliger, D. Kohn and H. Madry, “rAAV-Mediated Overexpression of FGF-2 Promotes Cell Proliferation, Survival, and Alpha-SMA Expression in Human Meniscal Lesions,” Gene Therapy, Vol. 16, No. 11, 2009, pp. 1363-1372.

[5]   S. Collier and P. Ghosh, “Effects of Transforming Growth Factor Beta on Proteoglycan Synthesis by Cell and Explant Cultures Derived from the Knee Joint Meniscus,” Osteoarthritis and Cartilage, Vol. 3, No. 2, 1995, pp. 127-138.

[6]   T. Tanaka, K. Fujii and Y. Kumagae, “Comparison of Biochemical Characteristics of Cultured Fibrochondrocytes Isolated from the Inner and Outer Regions of Hu- man Meniscus,” Knee Surgery, Sports Traumatology, Arthroscopy, Vol. 7, No. 2, 1999, pp. 75-80.

[7]   J. A. Buckwalter, H. J. Mankin and A. J. Grodzinsky, “Articular Cartilage and Osteoarthritis,” Instructional Course Lectures, Vol. 54, 2005, pp. 465-480.

[8]   M. W. Lark, E. K. Bayne, J. Flanagan, C. F. Harper, L. A. Hoerrner, N. I. Hutchinson, I. I. Singer, S. A. Donatelli, J. R. Weidner, H. R. Williams, R. A. Mumford and L. S. Lohmander, “Aggrecan Degradation in Human Cartilage. Evidence for Both Matrix Metalloproteinase and Aggrecanase Activity in Normal, Osteoarthritic, and Rheuma- toid Joints,” Journal of Clinical Investigation, Vol. 100, No. 1, 1997, pp. 93-106.

[9]   L. A. Fortier, H. O. Mohammed, G. Lust and A. J. Nixon, “Insulin-Like Growth Factor-I Enhances Cell-Based Repair of Articular Cartilage,” Journal of Bone & Joint Sur- gery, Vol. 84, No. 2, 2002, pp. 276-288.

[10]   S. B. Trippel, M. C. Whelan, M. Klagsbrun and S. R. Doctrow, “Interaction of Basic Fibroblast Growth Factor with Bovine Growth Plate Chondrocytes,” Journal of Orthopaedic Research, Vol. 10, No. 5, 1992, pp. 638-646.

[11]   F. D. Shuler, H. I. Georgescu, C. Niyibizi, R. K. Studer, Z. Mi, B. Johnstone, R. D. Robbins and C. H. Evans, “Increased Matrix Synthesis Following Adenoviral Transfer of a Transforming Growth Factor Beta1 Gene into Articular Chondrocytes,” Journal of Orthopaedic Research, Vol. 18, No. 4, 2000, pp. 585-592.

[12]   E. N. Blaney Davidson, E. L. Vitters, P. M. van der Kraan and W. B. van den Berg, “Expression of Transforming Growth Factor-Beta (TGFbeta) and the TGF-Beta Signalling Molecule SMAD-2P in Spontaneous and Instability-Induced Osteoarthritis: Role in Cartilage Degradation, Chondrogenesis and Osteophyte Formation,” Annals of the Rheumatic Diseases, Vol. 65, No. 11, 2006, pp. 1414-1421.

[13]   H. L. Moses and R. Serra, “Regulation of Differentiation by TGF-Beta,” Current Opinion in Genetics & Development, Vol. 6, No. 5, 1996, pp. 581-586.

[14]   P. A. Hoodless and J. L. Wrana, “Mechanism and Function of Signaling by the TGF Beta Superfamily,” Current Topics in Microbiology and Immunology, Vol. 228, 1998, pp. 235-272.

[15]   S. H. Tsai, M. T. Sheu, Y. C. Liang, H. T. Cheng, S. S. Fang and C. H. Chen, “TGF-Beta Inhibits IL-1beta-Activated PAR-2 Expression through Multiple Pathways in Human Primary Synovial Cells,” Journal of Biomedical Science, Vol. 16, 2009, p. 97.

[16]   G. J. Inman, F. J. Nicolás, J. F. Callahan, J. D. Harling, L. M. Gaster, A. D. Reith, N. J. Laping and C. S. Hill, “SB-431542 Is a Potent and Specific Inhibitor of Transforming Growth Factor-Beta Superfamily Type I Activin Receptor-Like Kinase (ALK) Receptors ALK4, ALK5, and ALK7,” Molecular Pharmacology, Vol. 62, No. 1, 2002, pp. 65-74.

[17]   N. J. Laping, E. Grygielko, A. Mathur, S. Butter, J. Bomberger, C. Tweed, W. Martin, J. Fornwald, R. Lehr, J. Harling, L. Gaster, J. F. Callahan and B. A. Olson, “Inhibition of Transforming Growth Factor (TGF)-Beta1-Induced Extracellular Matrix with a Novel Inhibitor of the TGF-Beta Type I Receptor Kinase Activity: SB-431542,” Molecular Pharmacology, Vol. 62, No. 1, 2002, pp. 58-64.

[18]   L. Longobardi, L. O’Rear, S. Aakula, B. Johnstone, K. Shimer, A. Chytil, W. A. Horton, H. L. Moses and A. Spagnoli, “Effect of IGF-I in the Chondrogenesis of Bone Marrow Mesenchymal Stem Cells in the Presence or Absence of TGF-Beta Signaling,” Journal of Bone and Mineral Research, Vol. 21, No. 4, 2006, pp. 626-636.

[19]   T. Tsukazaki, T. Usa, T. Matsumoto, H. Enomoto, A. Ohtsuru, H. Namba, K. Iwasaki and S. Yamashita, “Effect of Transforming Growth Factor-Beta on the Insulin-Like Growth Factor-I Autocrine/Paracrine Axis in Cultured Rat Articular Chondrocytes,” Experimental Cell Research, Vol. 215, No. 1, 1994, pp. 9-16.

[20]   K. T. Rousche, B. C. Ford, C. A. Praul and R. M. Leach, “The Use of Growth Factors in the Proliferation of Avian Articular Chondrocytes in a Serum-Free Culture System,” Connective Tissue Research, Vol. 42, No. 3, 2001, pp. 165-174.

[21]   J. D. Richmon, A. B. Sage, V. W. Wong, A. C. Chen, C. Pan, R. L. Sah and D. Watson, “Tensile Biomechanical Properties of Human Nasal Septal Cartilage,” American Journal of Rhinology, Vol. 19, No. 6, 2005, pp. 617-622.

[22]   M. Centrella, T. L. McCarthy and E. Canalis, “Trans- forming Growth Factor Beta Is a Bifunctional Regulator of Replication and Collagen Synthesis in Osteoblast-En- riched Cell Cultures from Fetal Rat Bone,” Journal of Biological Chemistry, Vol. 262, No. 6, 1987, pp. 2869- 2874.

[23]   M. K. Akens and M. B. Hurtig, “Influence of Species and Anatomical Location on Chondrocyte Expansion,” BMC Musculoskeletal Disorders, Vol. 6, 2005, p. 23.

[24]   A. Yonekura, M. Osaki, Y. Hirota, T. Tsukazaki, Y. Mi- yazaki, T. Matsumoto, A. Ohtsuru, H. Namba, H. Shindo and S. Yamashita, “Transforming Growth Factor-Beta Stimulates Articular Chondrocyte Cell Growth through p44/42 MAP Kinase (ERK) Activation,” Endocrine Journal, Vol. 46, No. 4, 1999, pp. 545-553.

[25]   G. Chen and N. Khalil, “TGF-Beta1 Increases Proliferation of Airway Smooth Muscle Cells by Phosphorylation of Map Kinases,” Respiratory Research, Vol. 7, 2006, p. 2.

[26]   S. Matsuyama, M. Iwadate, M. Kondo, M. Saitoh, A. Hanyu, K. Shimizu, H. Aburatani, H. K. Mishima, T. Imamura, K. Miyazono and K. Miyazawa, “SB-431542 and Gleevec Inhibit Transforming Growth Factor-Beta-Induced Proliferation of Human Osteosarcoma Cells,” Cancer Research, Vol. 63, No. 22, 2003, pp. 7791-7798.

[27]   H. Goto, F. D. Shuler, C. Niyibizi, F. H. Fu, P. D. Robbins and C. H. Evans, “Gene Therapy for Meniscal Injury: Enhanced Synthesis of Proteoglycan and Collagen by Meniscal Cells Transduced with a TGFbeta(1)Gene,” Osteoarthritis and Cartilage, Vol. 8, No. 4, 2000, pp. 266-271.

[28]   P. Galéra, D. Vivien, S. Pronost, J. Bonaventure, F. Rédini, G. Loyau and J. P. Pujol, “Transforming Growth Factor-Beta 1 (TGF-Beta 1) Up-Regulation of Collagen Type II in Primary Cultures of Rabbit Articular Chondrocytes (RAC) Involves Increased mRNA Levels without Affecting mRNA Stability and Procollagen Processing,” Journal of Cellular Physiology, Vol. 153, No. 3, 1992, pp. 596-606.

[29]   K. G. Vogel and D. J. Hernandez, “The Effects of Transforming Growth Factor-Beta and Serum on Proteoglycan Synthesis by Tendon Fibrocartilage,” European Journal of Cell Biology, Vol. 59, No. 2, 1992, pp. 304-313.