Physical Forces on Cells: Effects of Applied Osmotic Loading and Direct Current Electric Fields

Cartilage and the anterior cruciate ligament (ACL) exhibit poor healing capacity when injured. Cells from these orthopaedic tissues experience continuous changes in their chemical and physical environment that may potentially modulate their properties and behavior. This dissertation is the first characterization of the applied osmotic loading and direct current electric field response of chondrocytes and ACL fibroblasts. Osmotic loading was found to influence cell material properties, calcium signaling and gene expression. Applied direct current electric fields were shown to modulate and direct cell migration as well as wound healing. These lesser-studied physical forces were also found to have differential effects on cell behavior depending on whether they were applied statically or dynamically. The combined effects of these stimuli were also investigated and alterations in extracellular osmolarity were found to modulate chondrocyte migration behavior in applied electric fields. To study the novel effects of applied dynamic osmotic loading, a custom microfluidic device was developed to apply time-varying physical and chemical stimuli to cultured cells while permitting real-time monitoring of cellular responses. The experimental systems developed over the course of this biomedical engineering dissertation provide quantitative tools that can be used to gain further insights into the cellular response to osmotic loading and electric fields that may contribute new knowledge to our understanding of mechanotransduction as well as the role of physical forces in injury and repair of orthopaedic cells.

Year	Entry
2002	Davisson T, Kunig S, Chen A, Sah R, Ratcliffe A. Static and dynamic compression modulate matrix metabolism in tissue engineered cartilage. J Orthop Res. July 2002;20(4):842-848.
1977	Maroudas A, Venn M. Chemical composition and swelling of normal and osteoarthrotic femoral head cartilage, II: swelling. Ann Rheum Dis. 1977;36(5):399-406.
2001	Erickson GR, Alexopoulos LG, Guilak F. Hyper-osmotic stress induces volume change and calcium transients in chondrocytes by transmembrane, phospholipid, and G-protein pathways. J Biomech. December 2001;34(12):1527-1535.
1989	Urban JPG, Bayliss MT. Regulation of proteoglycan synthesis rate in cartilage in vitro: influence of extracellular ionic composition. Biochim Biophys Acta. July 21, 1989;992(1):59-65.
1994	Guilak F, Meyer BC, Ratcliffe A, Mow VC. The effects of matrix compression on proteoglycan metabolism in articular cartilage explants. Osteoarthritis Cartilage. June 1994;2(2):91-101.
1979	Maroudas A. Physicochemical properties of articular cartilage. In: Freeman MAR, ed. Adult Articular Cartilage. 2nd ed. Kent, England: Pitman Medical; 1979:215-290.
1995	Buschmann MD, Gluzband YA, Grodzinsky AJ, Hunziker EB. Mechanical compression modulates matrix biosynthesis in chondrocyte/agarose culture. J Cell Sci. April 1995;108(4):1497-1508.
2001	Obradovic B, Martin I, Padera RF, Treppo S, Freed LE, Vunjak-Navakovic G. Integration of engineered cartilage. J Orthop Res. November 2001;19(6):1089-1097.
2002	Wang CC-B, Guo XE, Sun D, Mow VC, Ateshian GA, Hung CT. The functional environment of chondrocytes within cartilage subjected to compressive loading: a theoretical and experimental approach. Biorheology. 2002;39(1-2):11-25.
1993	Urban JPG, Hall AC, Gehl KA. Regulation of matrix synthesis rates by the ionic and osmotic environment of articular chondrocytes. J Cell Physiol. February 1993;154(2):262-270.
1984	Palmoski MJ, Brandt KD. Effects of static and cyclic compressive loading on articular cartilage plugs in vitro. Arthritis Rheum. June 1984;27(6):675-681.
1995	Smith RL, Donlon BS, Gupta MK, Mohtai M, Das P, Carter DR, Cooke J, Gibbons G, Hutchinson N, Schurman DJ. Effects of fluid-induced shear on articular chondrocyte morphology and metabolism in vitro. J Orthop Res. November 1995;13(6):824-831.
1991	Ingber D. Integrins as mechanochemical transducers. Curr Opin Cell Biol. October 1991;3(5):841-848.
2002	Guilak F, Erickson GR, Ting-Beall HP. The effects of osmotic stress on the viscoelastic and physical properties of articular chondrocytes. Biophys J. February 2002;82(2):720-727.
1996	Hung CT, Allen FD, Pollack SR, Brighton CT. What is the role of the convective current density in the real-time calcium response of cultured bone cells to fluid flow? J Biomech. November 1996;29(11):1403-1409.
1993	Wang N, Butler JP, Ingber DE. Mechanotransduction across the cell surface and through the cytoskeleton. Science. May 21, 1993;260(5111):1124-1127.
1999	Guilak F, Ting-Beall HP, Baer AE, Trickey WR, Erickson GR, Setton LA. Viscoelastic properties of intervertebral disc cells: identification of two biomechanically distinct cell populations. Spine. December 1, 1999;24(23):2475-2483.
1989	Sah RL-Y, Kim Y-J, Doong J-YH, Grodzinsky AJ, Plass AHK, Sandy JD. Biosynthetic response of cartilage explants to dynamic compression. J Orthop Res. 1989;7(5):619-636.
1991	Lai WM, Hou JS, Mow VC. A triphasic theory for the swelling and deformation behaviors of articular cartilage. J Biomech Eng. August 1991;113(3):245-258.
1997	Wong M, Wuethrich P, Buschmann MD, Eggli P, Hunziker E. Chondrocyte biosynthesis correlates with local tissue strain in statically compressed adult articular cartilage. J Orthop Res. March 1997;15(2):189-196.
1999	Jones WR, Ping Ting-Beall H, Lee GM, Kelley SS, Hochmuth RM, Guilak F. Alterations in the young’s modulus and volumetric properties of chondrocytes isolated from normal and osteoarthritic human cartilage. J Biomech. February 1999;32(2):119-127.
1996	Hung CT, Allen FD, Pollack SR, Brighton CT. Intracellular Ca²⁺ stores and extracellular Ca²⁺ are required in the real-time Ca²⁺ response of bone cells experiencing fluid flow. J Biomech. November 1996;29(11):1411-1417.
1995	Reindel ES, Ayroso AM, Chen AC, Chun DM, Schinagl RM, Sah RL. Integrative repair of articular cartilage in vitro: adhesive strength of the interface region. J Orthop Res. September 1995;13(5):751-760.
2001	Roberts SR, Knight MM, Lee DA, Bader DL. Mechanical compression influences intracellular Ca²⁺ signaling in chondrocytes seeded in agarose constructs. J Appl Physiol. April 2001;90(4):1385-1391.
2000	Fortin M, Soulhat J, Shirazi-Adl A, Hunziker EB, Buschmann MD. Unconfined compression of articular cartilage: nonlinear behavior and comparison with a fibril-reinforced biphasic model. J Biomech Eng. April 2000;122(2):189-195.
1995	Buschmann MD, Grodzinsky AJ. A molecular model of proteoglycan-associated electrostatic forces in cartilage mechanics. J Biomech Eng. May 1995;117(2):179-192.
1999	Hunziker EB. Articular cartilage repair: are the intrinsic biological constraints undermining this process insuperable? Osteoarthritis Cartilage. January 1999;7(1):15-28.
1995	Guilak F, Ratcliffe A, Mow VC. Chondrocyte deformation and local tissue strain in articular cartilage: a confocal microscopy study. J Orthop Res. May 1995;13(3):410-421.
1995	Curnier A, He Q-C, Zysset P. Conewise linear elastic materials. J Elast. 1995;37(1):1-38.
1994	Urban JPG, Hall AC. The effects of hydrostatic and osmotic pressures on chondrocyte metabolism. In: Mow VC, Tran-Son-Tay R, Guilak F, Hochmuth RM, eds. Cell Mechanics and Cellular Engineering. Symposium on Cell Mechanics and Cellular Engineering; July 10-15, 1994. New York, NY: Springer-Verlag; 1994:398-419.
2000	Mauck RL, Soltz MA, Wang CCB, Wong DD, Chao P-HG, Valhmu WB, Hung CT, Ateshian GA. Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. J Biomech Eng. June 2000;122(3):252-260.
1962	Bassett CAL, Becker RO. Generation of electric potentials by bone in response to mechanical stress. Science. September 28, 1962;137(3535):1063.
1982	Mankin HJ. The response of articular cartilage to mechanical injury. J Bone Joint Surg. March 1982;64A(3):460-466.
1982	Hutton WC, Adams MA. Can the lumbar spine be crushed in heavy lifting? Spine. November 1982;7(6):586-590.
2004	Trickey WR, Vail TP, Guilak F. The role of the cytoskeleton in the viscoelastic properties of human articular chondrocytes. J Orthop Res. January 2004;22(1):131-139.
1995	Hung T Clark, Pollack R Solomon, Reilly TM, Brighton T Carl. Real-time calcium response of cultured bone cells to fluid flow. Clin Orthop Relat Res. April 1995;313:256-269.
2000	Hung CT, Ross Henshaw D, Wang CC-B, Mauck RL, Raia F, Palmer G, Grace Chao P-H, Mow VC, Ratcliffe A, Valhmu WB. Mitogen-activated protein kinase signaling in bovine articular chondrocytes in response to fluid flow does not require calcium mobilization. J Biomech. January 2000;33(1):73-80.
1994	Daniel DM, Stone ML, Dobson BE, Fithian DC, Rossman DJ, Kaufman KR. Fate of the ACL-injured patient: a prospective outcome study. Am J Sports Med. September–October 1994;22(5):632-644.
2001	Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods. December 2001;25(4):402-408.
2000	Butler DL, Goldstein SA, Guilak F. Functional tissue engineering: the role of biomechanics. J Biomech Eng. December 2000;122(6):570-575.
1991	Hall AC, Urban JPG, Gehl KA. The effects of hydrostatic pressure on matrix synthesis in articular cartilage. J Orthop Res. January 1991;9(1):1-10.
1970	Muir H, Bullough P, Maroudas A. The distribution of collagen in human articular cartilage with some of its physiological implications. J Bone Joint Surg. August 1970;52B(3):554-563.
2000	Soltz MA, Ateshian GA. A conewise linear elasticity mixture model for the analysis of tension-compression nonlinearity in articular cartilage. J Biomech Eng. December 2000;122(6):576-586.
1998	Valhmu WB, Stazzone EJ, Bachrach NM, Saed-Nejad F, Fischer SG, Mow VC, Ratcliffe A. Load-controlled compression of articular cartilage induces a transient stimulation of aggrecan gene expression. Arch Biochem Biophys. May 1, 1998;353(1):29-36.
1985	Levesque MJ, Nerem RM. The elongation and orientation of cultured endothelial cells in response to shear stress. J Biomech Eng. November 1985;107(4):341-347.
2004	Hung CT, Mauck RL, Wang CC-B, Lima EG, Ateshian GA. A paradigm for functional tissue engineering of articular cartilage via applied physiologic deformational loading. Ann Biomed Eng. January 2004;32(1):35-49.

Year

Entry

2002

Davisson T, Kunig S, Chen A, Sah R, Ratcliffe A. Static and dynamic compression modulate matrix metabolism in tissue engineered cartilage. J Orthop Res. July 2002;20(4):842-848.

1977

Maroudas A, Venn M. Chemical composition and swelling of normal and osteoarthrotic femoral head cartilage, II: swelling. Ann Rheum Dis. 1977;36(5):399-406.

2001

Erickson GR, Alexopoulos LG, Guilak F. Hyper-osmotic stress induces volume change and calcium transients in chondrocytes by transmembrane, phospholipid, and G-protein pathways. J Biomech. December 2001;34(12):1527-1535.

1989

Urban JPG, Bayliss MT. Regulation of proteoglycan synthesis rate in cartilage in vitro: influence of extracellular ionic composition. Biochim Biophys Acta. July 21, 1989;992(1):59-65.

1994

Guilak F, Meyer BC, Ratcliffe A, Mow VC. The effects of matrix compression on proteoglycan metabolism in articular cartilage explants. Osteoarthritis Cartilage. June 1994;2(2):91-101.