Synovial fibroblasts self-direct multicellular lining architecture and synthetic function in three-dimensional organ culture
Gerald F. M. Watts
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorJohn Wright
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorThomas S. Thornhill
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorMarkus Sköld
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorSamuel M. Behar
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorBirgit Niederreiter
Medical University of Vienna, Vienna, Austria
Search for more papers by this authorManuela Cernadas
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorAnthony J. Coyle
MedImmune, Inc., Gaithersburg, Maryland
Drs. Coyle and Sims own stock or stock options in MedImmune/AstraZeneca.
Search for more papers by this authorGary P. Sims
MedImmune, Inc., Gaithersburg, Maryland
Drs. Coyle and Sims own stock or stock options in MedImmune/AstraZeneca.
Search for more papers by this authorMatthew L. Warman
Children's Hospital Boston, Boston, Massachusetts
Search for more papers by this authorMichael B. Brenner
Brigham and Women's Hospital, Boston, Massachusetts
Dr. Brenner has received consulting fees from Synovex (more than $10,000), owns stock options in Synovex, and holds patents on cadherin-11, licensed to Synovex.
Search for more papers by this authorCorresponding Author
David M. Lee
Brigham and Women's Hospital, Boston, Massachusetts
Dr. Lee has received consulting fees from UCB, Astellas, and Resolvyx (less than $10,000 each) as well as from Synovex (more than $10,000), he owns stock or stock options in Synovex, and together with Synovex, he holds a patent for anti–cadherin 11 (also expressed on fibroblast-like synoviocytes).
Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital, Harvard Medical School, Smith 552, 1 Jimmy Fund Way, Boston, MA 02115Search for more papers by this authorGerald F. M. Watts
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorJohn Wright
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorThomas S. Thornhill
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorMarkus Sköld
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorSamuel M. Behar
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorBirgit Niederreiter
Medical University of Vienna, Vienna, Austria
Search for more papers by this authorManuela Cernadas
Brigham and Women's Hospital, Boston, Massachusetts
Search for more papers by this authorAnthony J. Coyle
MedImmune, Inc., Gaithersburg, Maryland
Drs. Coyle and Sims own stock or stock options in MedImmune/AstraZeneca.
Search for more papers by this authorGary P. Sims
MedImmune, Inc., Gaithersburg, Maryland
Drs. Coyle and Sims own stock or stock options in MedImmune/AstraZeneca.
Search for more papers by this authorMatthew L. Warman
Children's Hospital Boston, Boston, Massachusetts
Search for more papers by this authorMichael B. Brenner
Brigham and Women's Hospital, Boston, Massachusetts
Dr. Brenner has received consulting fees from Synovex (more than $10,000), owns stock options in Synovex, and holds patents on cadherin-11, licensed to Synovex.
Search for more papers by this authorCorresponding Author
David M. Lee
Brigham and Women's Hospital, Boston, Massachusetts
Dr. Lee has received consulting fees from UCB, Astellas, and Resolvyx (less than $10,000 each) as well as from Synovex (more than $10,000), he owns stock or stock options in Synovex, and together with Synovex, he holds a patent for anti–cadherin 11 (also expressed on fibroblast-like synoviocytes).
Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital, Harvard Medical School, Smith 552, 1 Jimmy Fund Way, Boston, MA 02115Search for more papers by this authorAbstract
Objective
To define the intrinsic capacity of fibroblast-like synoviocytes (FLS) to establish a 3-dimensional (3-D) complex synovial lining architecture characterized by the multicellular organization of the compacted synovial lining and the elaboration of synovial fluid constituents.
Methods
FLS were cultured in spherical extracellular matrix (ECM) micromasses for 3 weeks. The FLS micromass architecture was assessed histologically and compared with that of dermal fibroblast controls. Lubricin synthesis was measured via immunodetection. Basement membrane matrix and reticular fiber stains were performed to examine ECM organization. Primary human and mouse monocytes were prepared and cocultured with FLS in micromass to investigate cocompaction in the lining architecture. Cytokine stimuli were applied to determine the capacity for inflammatory architecture rearrangement.
Results
FLS, but not dermal fibroblasts, spontaneously formed a compacted lining architecture over 3 weeks in the 3-D ECM micromass organ cultures. These lining cells produced lubricin. FLS rearranged their surrounding ECM into a complex architecture resembling the synovial lining and supported the survival and cocompaction of monocyte/macrophages in the neo–lining structure. Furthermore, when stimulated by cytokines, FLS lining structures displayed features of the hyperplastic rheumatoid arthritis synovial lining.
Conclusion
This 3-D micromass organ culture method demonstrates that many of the phenotypic characteristics of the normal and the hyperplastic synovial lining in vivo are intrinsic functions of FLS. Moreover, FLS promote survival and cocompaction of primary monocytes in a manner remarkably similar to that of synovial lining macrophages. These findings provide new insight into inherent functions of the FLS lineage and establish a powerful in vitro method for further investigation of this lineage.
REFERENCES
- 1 Castor CW. The microscopic structure of normal human synovial tissue. Arthritis Rheum 1960; 3: 140–51.
- 2 Barland P, Novikoff AB, Hamerman D. Electron microscopy of the human synovial membrane. J Cell Biol 1962; 14: 207–20.
- 3 Lee DM, Kiener HP, Brenner MB. Synoviocytes. In: ED Harris, RC Budd, GS Firestein, MC Genovese, JS Sergent, S Ruddy, CB Sledge, editors. Kelley's textbook of rheumatology. 7th ed. Philadelphia: Saunders Elsevier; 2004. p. 175–88.
- 4 Lever JD, Ford EH. Histological, histochemical and electron microscopic observations on synovial membrane. Anat Rec 1958; 132: 525–39.
- 5 Revell PA, al-Saffar N, Fish S, Osei D. Extracellular matrix of the synovial intimal cell layer. Ann Rheum Dis 1995; 54: 404–7.
- 6 Mapp PI, Revell PA. Fibronectin production by synovial intimal cells. Rheumatol Int 1985; 5: 229–37.
- 7 Pollock LE, Lalor P, Revell PA. Type IV collagen and laminin in the synovial intimal layer: an immunohistochemical study. Rheumatol Int 1990; 9: 277–80.
- 8 Ashhurst DE, Bland YS, Levick JR. An immunohistochemical study of the collagens of rabbit synovial interstitium. J Rheumatol 1991; 18: 1669–72.
- 9 Wolf J, Carsons SE. Distribution of type VI collagen expression in synovial tissue and cultured synoviocytes: relation to fibronectin expression. Ann Rheum Dis 1991; 50: 493–6.
- 10 Smith SC, Folefac VA, Osei DK, Revell PA. An immunocytochemical study of the distribution of proline-4-hydroxylase in normal, osteoarthritic and rheumatoid arthritic synovium at both the light and electron microscopic level. Br J Rheumatol 1998; 37: 287–91.
- 11 McCachren SS, Lightner VA. Expression of human tenascin in synovitis and its regulation by interleukin-1. Arthritis Rheum 1992; 35: 1185–96.
- 12 Danen EH, Sonneveld P, Brakebusch C, Fassler R, Sonnenberg A. The fibronectin-binding integrins α5β1 and αvβ3 differentially modulate RhoA-GTP loading, organization of cell matrix adhesions, and fibronectin fibrillogenesis. J Cell Biol 2002; 159: 1071–86.
- 13 Okada Y, Nagase H, Harris ED Jr. Matrix metalloproteinases 1, 2, and 3 from rheumatoid synovial cells are sufficient to destroy joints. J Rheumatol 1987; 14: 41–2.
- 14 Woessner JF Jr. Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 1991; 5: 2145–54.
- 15 Konttinen YT, Ainola M, Valleala H, Ma J, Ida H, Mandelin J, et al. Analysis of 16 different matrix metalloproteinases (MMP-1 to MMP-20) in the synovial membrane: different profiles in trauma and rheumatoid arthritis. Ann Rheum Dis 1999; 58: 691–7.
- 16 Close DR. Matrix metalloproteinase inhibitors in rheumatic diseases. Ann Rheum Dis 2001; 60 Suppl 3: iii62–7.
- 17 Wojtowicz-Praga S, Torri J, Johnson M, Steen V, Marshall J, Ness E, et al. Phase I trial of marimastat, a novel matrix metalloproteinase inhibitor, administered orally to patients with advanced lung cancer. J Clin Oncol 1998; 16: 2150–6.
- 18 Rasmussen HS, McCann PP. Matrix metalloproteinase inhibition as a novel anticancer strategy: a review with special focus on batimastat and marimastat. Pharmacol Ther 1997; 75: 69–75.
- 19 Nemunaitis J, Poole C, Primrose J, Rosemurgy A, Malfetano J, Brown P, et al. Combined analysis of studies of the effects of the matrix metalloproteinase inhibitor marimastat on serum tumor markers in advanced cancer: selection of a biologically active and tolerable dose for longer-term studies. Clin Cancer Res 1998; 4: 1101–9.
- 20 Yielding KL, Tomkins GM, Bunim JJ. Synthesis of hyaluronic acid by human synovial tissue slices. Science 1957; 125: 1300.
- 21 Wilkinson LS, Pitsillides AA, Worrall JG, Edwards JC. Light microscopic characterization of the fibroblast-like synovial intimal cell (synoviocyte). Arthritis Rheum 1992; 35: 1179–84.
- 22 Pitsillides AA, Blake SM. Uridine diphosphoglucose dehydrogenase activity in synovial lining cells in the experimental antigen induced model of rheumatoid arthritis: an indication of synovial lining cell function. Ann Rheum Dis 1992; 51: 992–5.
- 23 Pitsillides AA, Wilkinson LS, Mehdizadeh S, Bayliss MT, Edwards JC. Uridine diphosphoglucose dehydrogenase activity in normal and rheumatoid synovium: the description of a specialized synovial lining cell. Int J Exp Pathol 1993; 74: 27–34.
- 24 Rhee DK, Marcelino J, Baker M, Gong Y, Smits P, Lefebvre V, et al. The secreted glycoprotein lubricin protects cartilage surfaces and inhibits synovial cell overgrowth. J Clin Invest 2005; 115: 622–31.
- 25 Henderson B, Pettipher ER. The synovial lining cell: biology and pathobiology. Semin Arthritis Rheum 1985; 15: 1–32.
- 26 Jay GD, Britt DE, Cha CJ. Lubricin is a product of megakaryocyte stimulating factor gene expression by human synovial fibroblasts. J Rheumatol 2000; 27: 594–600.
- 27 Swann DA, Silver FH, Slayter HS, Stafford W, Shore E. The molecular structure and lubricating activity of lubricin isolated from bovine and human synovial fluids. Biochem J 1985; 225: 195–201.
- 28 Zvaifler NJ, Tsai V, Alsalameh S, von Kempis J, Firestein GS, Lotz M. Pannocytes: distinctive cells found in rheumatoid arthritis articular cartilage erosions. Am J Pathol 1997; 150: 1125–38.
- 29 Lee DM, Kiener HP, Agarwal SK, Noss EH, Watts GF, Chisaka O, et al. Cadherin-11 in synovial lining formation and pathology in arthritis. Science 2007; 315: 1006–10.
- 30 Bucala R, Ritchlin C, Winchester R, Cerami A. Constitutive production of inflammatory and mitogenic cytokines by rheumatoid synovial fibroblasts. J Exp Med 1991; 173: 569–74.
- 31 Ritchlin C. Fibroblast biology: effector signals released by the synovial fibroblast in arthritis. Arthritis Res 2000; 2: 356–60.
- 32 Firestein GS. Invasive fibroblast-like synoviocytes in rheumatoid arthritis: passive responders or transformed aggressors? [review]. Arthritis Rheum 1996; 39: 1781–90.
- 33 Kiener HP, Lee DM, Agarwal SK, Brenner MB. Cadherin-11 induces rheumatoid arthritis fibroblast-like synoviocytes to form lining layers in vitro. Am J Pathol 2006; 168: 1486–99.
- 34 Kouskoff V, Korganow AS, Duchatelle V, Degott C, Benoist C, Mathis D. Organ-specific disease provoked by systemic autoimmunity. Cell 1996; 87: 811–22.
- 35 Brigl M, van den Elzen P, Chen X, Meyers JH, Wu D, Wong CH, et al. Conserved and heterogeneous lipid antigen specificities of CD1d-restricted NKT cell receptors. J Immunol 2006; 176: 3625–34.
- 36 Skold M, Behar SM. Tuberculosis triggers a tissue-dependent program of differentiation and acquisition of effector functions by circulating monocytes. J Immunol 2008; 181: 6349–60.
- 37 Sarmiento M, Loken MR, Fitch FW. Structural differences in cell surface T25 polypeptides from thymocytes and cloned T cells. Hybridoma 1981; 1: 13–26.
- 38 Humason GL. Animal tissue techniques. 2nd ed. New York: WH Freeman; 1967.
- 39 Carson FL. Histotechnology. Chicago: ASCP Press; 1997.
- 40 Smith S. Fifteen-minute Jones' stain for renal biopsies. Lab Med 1979; 10: 769–71.
- 41 JD Bancroft, A Stevens, editors. Theory and practice of histological techniques. 4th ed. New York: Churchill Livingstone; 1996.
- 42 Rhee DK, Marcelino J, Al-Mayouf S, Schelling DK, Bartels CF, Cui Y, et al. Consequences of disease-causing mutations on lubricin protein synthesis, secretion, and post-translational processing. J Biol Chem 2005; 280: 31325–32.
- 43 Myers DB, Highton TC, Rayns DG. Acid mucopolysaccharides closely associated with collagen fibrils in normal human synovium. J Ultrastruct Res 1969; 28: 203–13.
- 44 Eyre DR, Muir H. Type III collagen: a major constituent of rheumatoid and normal human synovial membrane. Connect Tissue Res 1975; 4: 11–6.
- 45 Scott DL, Salmon M, Morris CJ, Wainwright AC, Walton KW. Laminin and vascular proliferation in rheumatoid arthritis. Ann Rheum Dis 1984; 43: 551–5.
- 46 Clemmensen I, Holund B, Andersen RB. Fibrin and fibronectin in rheumatoid synovial membrane and rheumatoid synovial fluid. Arthritis Rheum 1983; 26: 479–85.
- 47 Iwai-Liao Y, Ogita Y, Tsubai T, Higashi Y. Scanning electron microscopy of elastic system fibers in the articular disc of the rat mandibular joint. Okajimas Folia Anat Jpn 1994; 71: 211–25.
- 48 Konttinen YT, Li TF, Xu JW, Tagaki M, Pirila L, Silvennoinen T, et al. Expression of laminins and their integrin receptors in different conditions of synovial membrane and synovial membrane-like interface tissue. Ann Rheum Dis 1999; 58: 683–90.
- 49 B Henderson, JC Edwards, editors. The synovial lining in health and disease. London: Chapman & Hall; 1987.
- 50 Waggett AD, Kielty CM, Shuttleworth CA. Microfibrillar elements in the synovial joint: presence of type VI collagen and fibrillin-containing microfibrils. Ann Rheum Dis 1993; 52: 449–53.
- 51 Worrall JG, Wilkinson LS, Bayliss MT, Edwards JC. Zonal distribution of chondroitin-4-sulphate/dermatan sulphate and chondroitin-6-sulphate in normal and diseased human synovium. Ann Rheum Dis 1994; 53: 35–8.
- 52 Schneider M, Voss B, Rauterberg J, Menke M, Pauly T, Miehlke RK, et al. Basement membrane proteins in synovial membrane: distribution in rheumatoid arthritis and synthesis by fibroblast-like cells. Clin Rheumatol 1994; 13: 90–7.
- 53 Linck G, Stocker S, Grimaud JA, Porte A. Distribution of immunoreactive fibronectin and collagen (type I, III, IV) in mouse joints: fibronectin, an essential component of the synovial cavity border. Histochemistry 1983; 77: 323–8.
- 54 Marsh JM, Wiebkin OW, Gale S, Muir H, Maini RN. Synthesis of sulphated proteoglycans by rheumatoid and normal synovial tissue in culture. Ann Rheum Dis 1979; 38: 166–70.
- 55 Demaziere A, Athanasou NA. Adhesion receptors of intimal and subintimal cells of the normal synovial membrane. J Pathol 1992; 168: 209–15.
- 56 Okada Y, Naka K, Minamoto T, Ueda Y, Oda Y, Nakanishi I, et al. Localization of type VI collagen in the lining cell layer of normal and rheumatoid synovium. Lab Invest 1990; 63: 647–56.
- 57 Ushiki T. Collagen fibers, reticular fibers and elastic fibers: a comprehensive understanding from a morphological viewpoint. Arch Histol Cytol 2002; 65: 109–26.
- 58 Montes GS, Krisztan RM, Shigihara KM, Tokoro R, Mourao PA, Junqueira LC. Histochemical and morphological characterization of reticular fibers. Histochemistry 1980; 65: 131–41.
- 59 Athanasou NA, Quinn J. Immunocytochemical analysis of human synovial lining cells: phenotypic relation to other marrow derived cells. Ann Rheum Dis 1991; 50: 311–5.
- 60 Edwards JC, Willoughby DA. Demonstration of bone marrow derived cells in synovial lining by means of giant intracellular granules as genetic markers. Ann Rheum Dis 1982; 41: 177–82.
- 61 Cline MJ, Lehrer RI, Territo MC, Golde DW. Monocytes and macrophages: functions and diseases. Ann Intern Med 1978; 88: 78–88.
