Abstract
Diffusive transport of solutes is critical to the normal function of articular cartilage. The diffusion of macromolecules through cartilage may be affected by the local composition and structure, which vary with depth from the tissue surface. We hypothesized that the diffusion coefficient of uncharged molecules also varies with depth and molecular size. We used fluorescence recovery after photobleaching (FRAP) to measure site-specific diffusion coefficients of fluorescent dextran molecules (3, 40, 70, and 500 kDa) in porcine articular cartilage. The diffusion coefficients measured using FRAP exhibited an inverse size dependence and were in general agreement with those measured using other techniques. The diffusion coefficients for all molecules varied significantly with depth in a manner that depended upon the size of the diffusing molecule. The diffusion coefficients for the 3 and 500 kDa dextrans were 1.6 and 2.4 times greater, respectively, in the surface zone as compared to the middle and deep zones, whereas the diffusion coefficients of the 40 and 70 kDa dextrans were 0.3 and 0.2 times lower in the surface zone as compared to the middle and deep zones. These differences may reflect variations in the structure and composition of collagen, proteoglycans, and other macromolecules among the zones.© 2003 Biomedical Engineering Society.
PAC2003: 8715Vv, 8719Rr, 8764Tt
Similar content being viewed by others
References
Alexopoulos, L. G, M. A. Haider, T. P. Vail, and F. Guilak. Alterations in the mechanical properties of the human chondrocyte pericellular matrix with osteoarthritis. J. Biomech. Eng125:323–333, 2003.
Axelrod, D, D. E. Koppel, J. Schlessinger, E. Elson, and W. W. Webb. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys. J16:1055–1069, 1976.
Bayliss, M. T, M. Venn, A. Maroudas, and S. Y. Ali. Structure of proteoglycans from different layers of human articular cartilage. Biochem. J209:387–400, 1983.
Blonk, J. C. G, A. Don, H. Van Aalst, and J. J. Birmingham. Fluorescence photobleaching recovery in the confocal scanning light microscope. J. Microsc169:363–374, 1993.
Bryers, J. D, and F. Drummond. Local macromolecule diffusion coefficients in structurally non-uniform bacterial biofilms using fluorescence recovery after photobleaching (FRAP). Biotechnol. Bioeng60:462–473, 1998.
Burstein, D, M. L. Gray, A. L. Hartman, R. Gipe, and B. D. Foy. Diffusion of small solutes in cartilage as measured by nuclear magnetic resonance (NMR) spectroscopy and imaging. J. Orthop. Res11:465–478, 1993.
Clague, D. S, and R. J. Phillips. Hindered diffusion of spherical macromolecules through dilute fibrous media. Phys. Fluids8:1720–1731, 1996.
Comper, W. D Physicochemical aspects of cartilage extracellular matrix. In: Cartilage: Molecular Aspects, edited by B. Hall and S. Newman. Boston: CRC Press, 1991, pp. 59–96.
Eggli, P. S, E. B. Hunziker, and R. K. Schenk. Quantitation of structural features characterizing weight-and less-weight-bearing regions in articular cartilage: A stereological analysis of medial femoral condyles in young adult rabbits. Anat. Rec222:217–227, 1988.
Fischer, A. E, T. A. Carpenter, J. A. Tyler, and L. D. Hall. Visualization of mass transport of small organic molecules and metal ions through articular cartilage by magnetic resonance imaging. Magn. Reson. Imaging13:819–826, 1995.
Fischer, A. E, and L. D. Hall. Visualization of the diffusion of metal ions and organic molecules by magnetic resonance imaging of water. Magn. Reson. Imaging14:779–783, 1996.
Franzen, A, S. Inerot, S. O. Hejderup, and D. Heinegard. Variations in the composition of bovine hip articular cartilage with distance from the articular surface. Biochem. J195:535–543, 1981.
Gribbon, P, and T. E. Hardingham. Macromolecular diffusion of biological polymers measured by confocal fluorescence recovery after photobleaching. Biophys. J75:1032–1039, 1998.
Guilak, F, A. Ratcliffe, N. Lane, M. P. Rosenwasser, and V. C. Mow. Mechanical and biochemical changes in the superficial zone of articular cartilage in canine experimental osteoarthritis. J. Orthop. Res.12:474–484, 1994.
Han, J, and J. Herzfeld. Macromolecular diffusion in crowded solutions. Biophys. J65:1155–1161, 1993.
Hardingham, T. E, A. J. Fosang, and J. Dudhia. The structure, function and turnover of aggrecan, the large aggregating proteoglycan from cartilage. Eur. J. Clin. Chem. Clin. Biochem32:249–257, 1994.
Heinegard, D, and A. Oldberg. Structure and biology of cartilage and bone matrix noncollagenous macromolecules. FASEB J3:2042–2051, 1989.
Hunziker, E. B., M. Michel, and D. Studer. Ultrastructure of adult human articular cartilage matrix after cryotechnical processing. Microsc. Res. Tech37:271–284, 1997.
Hwang, W. S, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. Hughes. Collagen fibril structure of normal, aging, and osteoarthritic cartilage. J. Pathol167:425–433, 1992.
Jeffery, A. K, G. W. Blunn, C. W. Archer, and G. Bentley. Three-dimensional collagen architecture in bovine articular cartilage. J. Bone Jt. Surg, Br. Vol73:795–801, 1991.
Lang, I, M. Scholz, and R. Peters. Molecular mobility and nucleocytoplasmic flux in hepatoma cells. J. Cell Biol102:1183–1190, 1986.
Langsjo, T. K, M. Hyttinen, A. Pelttari, K. Kiraly, J. Arokoski, and H. J. Helminen. Electron microscopic stereological study of collagen fibrils in bovine articular cartilage: Volume and surface densities are best obtained indirectly (from length densities and diameters) using isotropic uniform random sampling. J. Anat195:281–293, 1999.
Lee, G. M, T. A. Paul, M. Slabaugh, and S. S. Kelley. The incidence of enlarged chondrons in normal and osteoarthritic human cartilage and their relative matrix density. Osteoarthritis Cartilage8:44–52, 2000.
Lipshitz, H, R. Etheredge, III, and M. J. Glimcher. Changes in the hexosamine content and swelling ratio of articular cartilage as functions of depth from the surface. J. Bone Jt. Surg58:1149–1153, 1976.
Luby-Phelps, K, D. L. Taylor, and F. Lanni. Probing the structure of cytoplasm. J. Cell Biol102:2015–2022, 1986.
Maroudas, A. Distribution and diffusion of solutes in articular cartilage. Biophys. J10:365–379, 1970.
Maroudas, A. Biophysical chemistry of cartilaginous tissues with special reference to solute and fluid transport. Biorheology12:233–248, 1975.
Maroudas, A. Physicochemical properties of articular cartilage. In: Adult Articular Cartilage, edited by M. A. R. Freeman. Bath: Pitman Medical, 1979, pp. 215–290
Netti, P. A, D. A. Berk, M. A. Swartz, A. J. Grodzinsky, and R. K. Jain. Role of extracellular matrix assembly in interstitial transport in solid tumors. Cancer Res60:2497–2503, 2000.
O'Hara, B. P, J. P. Urban, and A. Maroudas. Influence of cyclic loading on the nutrition of articular cartilage. Ann. Rheum. Dis49:536–539, 1990.
Papadopoulos, S, V. Endeward, B. Revesz-Walker, K. D. Jurgens, and G. Gros. Radial and longitudinal diffusion of myoglobin in single living heart and skeletal muscle cells. Proc. Natl. Acad. Sci. U.S.A98:5904–5909, 2001.
Pluen, A, Y. Boucher, S. Ramanujan, T. D. McKee, T. Gohongi, E. di Tomaso, E. B. Brown, Y. Izumi, R. B. Campbell, D. A. Berk, and R. K. Jain. Role of tumor-host interactions in interstitial diffusion of macromolecules: Cranial vs. subcutaneous tumors. Proc. Natl. Acad. Sci. U.S.A98:4628–4633, 2001.
Pluen, A, P. A. Netti, R. K. Jain, and D. A. Berk. Diffusion of macromolecules in agarose gels: Comparison of linear and globular configurations. Biophys. J77:542–552, 1999.
Poole, C. A, A. Matsuoka, and J. R. Schofield. Chondrons from articular cartilage. III. Morphologic changes in the cellular microenvironment of chondrons isolated from osteoarthritic cartilage. Arthritis Rheum34:22–35, 1991.
Quinn, T. M, P. Kocian, and J. J. Meister. Static compression is associated with decreased diffusivity of dextrans in cartilage explants. Arch. Biochem. Biophys384:327–334, 2000.
Seksek, O, J. Biwersi, and A. S. Verkman. Translational diffusion of macromolecule-sized solutes in cytoplasm and nucleus. J. Cell Biol138:131–142, 1997.
Seyedin, S. M, and D. M. Rose. Cartilage growth and differentiation factors. In: Cartilage: Molecular Aspects, edited by B. Hall and S. Newman. Boston: CRC Press, 1991, pp. 131–151
Smith, B. A, W. R. Clark, and H. M. McConnell. Anisotropic molecular motion on cell surfaces. Proc. Natl. Acad. Sci. U.S.A76:5641–5644, 1979.
Torzilli, P. A. Effects of temperature, concentration and articular surface removal on transient solute diffusion in articular cartilage. Med. Biol. Eng. Comput31:S93–98, 1993.
Torzilli, P. A, T. C. Adams, and R. J. Mis. Transient solute diffusion in articular cartilage. J. Biomech20:203–214, 1987.
Torzilli, P. A, J. M. Arduino, J. D. Gregory, and M. Bansal. Effect of proteoglycan removal on solute mobility in articular cartilage. J. Biomech30:895–902, 1997.
Torzilli, P. A, D. A. Grande, and J. M. Arduino. Diffusive properties of immature articular cartilage. J. Biomed. Mater. Res40:132–138, 1998.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Leddy, H.A., Guilak, F. Site-Specific Molecular Diffusion in Articular Cartilage Measured using Fluorescence Recovery after Photobleaching. Annals of Biomedical Engineering 31, 753–760 (2003). https://doi.org/10.1114/1.1581879
Issue Date:
DOI: https://doi.org/10.1114/1.1581879