0
Research Papers

The Regional Sensitivity of Chondrocyte Gene Expression to Coactive Mechanical Load and Exogenous TNF-α Stimuli

[+] Author and Article Information
S. L. Bevill

Mem. ASME
Department of Physical and
Environmental Sciences,
Colorado Mesa University,
1100 North Avenue,
Grand Junction, CO 81501
e-mail: sbevill@coloradomesa.edu

K. A. Boyer

Department of Kinesiology,
University of Massachusetts-Amherst,
110 Totman Building,
30 Eastman Lane,
Amherst, MA 01003-9258
e-mail: kboyer@kin.umass.edu

T. P. Andriacchi

Department of Mechanical Engineering,
Stanford University,
227 Durand Building,
Stanford, CA 94305-4038;
Department of Orthopedic Surgery,
Stanford University Medical Center,
227 Durand Building,
Stanford, CA 94305-4038;
Bone and Joint Center of Excellence,
Palo Alto, CA
e-mail: tandriac@stanford.edu

1Corresponding author.

Manuscript received December 18, 2013; final manuscript received June 18, 2014; accepted manuscript posted July 2, 2014; published online July 15, 2014. Assoc. Editor: Carlijn V. C. Bouten.

J Biomech Eng 136(9), 091005 (Jul 15, 2014) (7 pages) Paper No: BIO-13-1579; doi: 10.1115/1.4027937 History: Received December 18, 2013; Revised June 18, 2014; Accepted July 02, 2014

Both mechanical load and elevated levels of proinflammatory cytokines have been associated with the risk for developing osteoarthritis (OA), yet the potential interaction of these mechanical and biological factors is not well understood. The purpose of this study was to evaluate the response of chondrocytes to the effects of dynamic unconfined compression, TNF-α, and the simultaneous effects of dynamic unconfined compression and TNF-α. The response to these three treatments was markedly different and, taken together, the response in the gene expression of chondrocytes to the different treatment conditions suggest a complex interaction between structure, biology, and mechanical loading.

FIGURES IN THIS ARTICLE
<>
Copyright © 2014 by ASME
Your Session has timed out. Please sign back in to continue.

References

Kannus, P., and Jarvinen, M., 1989, “Posttraumatic Anterior Cruciate Ligament Insufficiency as a Cause of Osteoarthritis in a Knee Joint,” Clin. Rheumatol, 8(2), pp. 251–260. [CrossRef] [PubMed]
Lohmander, L. S., Englund, P. M., Dahl, L. L., and Roos, E. M., 2007, “The Long-Term Consequence of Anterior Cruciate Ligament and Meniscus Injuries: Osteoarthritis,” Am. J. Sports Med., 35(10), pp. 1756–1769. [CrossRef] [PubMed]
Roos, H., Adalberth, T., Dahlberg, L., and Lohmander, L. S., 1995, “Osteoarthritis of the Knee After Injury to the Anterior Cruciate Ligament or Meniscus: The Influence of Time and Age,” Osteoarthritis Cartilage, 3(4), pp. 261–267. [CrossRef] [PubMed]
Andriacchi, T. P., and Dyrby, C. O., 2005, “Interactions Between Kinematics and Loading During Walking for the Normal and ACL Deficient Knee,” J. Biomech., 38(2), pp. 293–298. [CrossRef] [PubMed]
Li, G., Moses, J. M., Papannagari, R., Pathare, N. P., DeFrate, L. E., and Gill, T. J., 2006, “Anterior Cruciate Ligament Deficiency Alters the in Vivo Motion of the Tibiofemoral Cartilage Contact Points in Both the Anteroposterior and Mediolateral Directions,” J. Bone Jt. Surg. Am., 88(8), pp. 1826–1834. [CrossRef]
Scanlan, S. F., Chaudhari, A. M., Dyrby, C. O., and Andriacchi, T. P., 2010, “Differences in Tibial Rotation During Walking in ACL Reconstructed and Healthy Contralateral Knees,” J. Biomech., 43(9), pp. 1817–1822. [CrossRef] [PubMed]
Cameron, M. L., Fu, F. H., Paessler, H. H., Schneider, M., and Evans, C. H., 1994, “Synovial Fluid Cytokine Concentrations as Possible Prognostic Indicators in the ACL-Deficient Knee,” Knee Surg. Sports Traumatol. Arthroscopy, 2(1), pp. 38–44. [CrossRef]
Cameron, M., Buchgraber, A., Passler, H., Vogt, M., Thonar, E., and Fu, F., 1997, “The Natural History of the Anterior Cruciate Ligament-Deficient Knee. Changes in Synovial Fluid Cytokine and Keratan Sulfate Concentrations,” Am. J. Sports Med., 25(6), pp. 751–754. [CrossRef] [PubMed]
Dahlberg, L., Friden, T., Roos, H., Lark, M. W., and Lohmander, L. S., 1994, “A Longitudinal Study of Cartilage Matrix Metabolism in Patients With Cruciate Ligament Rupture—Synovial Fluid Concentrations of Aggrecan Fragments, Stromelysin-1 and Tissue Inhibitor of Metalloproteinase-1,” Br. J. Rheumatol., 33(12), pp. 1107–1111. [CrossRef] [PubMed]
Irie, K., Uchiyama, E., and Iwaso, H., 2003, “Intraarticular Inflammatory Cytokines in Acute Anterior Cruciate Ligament Injured Knee,” Knee, 10(1), pp. 93–96. [CrossRef] [PubMed]
Higuchi, H., Shirakura, K., Kimura, M., Terauchi, M., Shinozaki, T., and Watanabe, H., 2006, “Changes in Biochemical Parameters After Anterior Cruciate Ligament Injury,” Int. Orthop., 30(1), pp. 43–47. [CrossRef] [PubMed]
Marks, P. H., and Donaldson, M. L., 2005, “Inflammatory Cytokine Profiles Associated With Chondral Damage in the Anterior Cruciate Ligament-Deficient Knee,” Arthroscopy, 21(11), pp. 1342–1347. [CrossRef] [PubMed]
Andriacchi, T. P., Mundermann, A., Smith, R. L., Alexander, E. J., Dyrby, C. O., and Koo, S., 2004, “A Framework for the in Vivo Pathomechanics of Osteoarthritis at the Knee,” Ann. Biomed. Eng., 32(3), pp. 447–457. [CrossRef] [PubMed]
Andriacchi, T. P., Koo, S., and Scanlan, S. F., 2009, “Gait Mechanics Influence Healthy Cartilage Morphology and Osteoarthritis of the Knee,” J. Bone Jt. Surg. Am., 91(Suppl. 1), pp. 95–101. [CrossRef]
Chaudhari, A. M., Briant, P. L., Bevill, S. L., Koo, S., and Andriacchi, T. P., 2008, “Knee Kinematics, Cartilage Morphology, and Osteoarthritis After ACL Injury,” Med. Sci. Sports Exercise, 40(2), pp. 215–222. [CrossRef]
Bevill, S. L., Briant, P. L., Levenston, M. E., and Andriacchi, T. P., 2009, “Central and Peripheral Region Tibial Plateau Chondrocytes Respond Differently to in Vitro Dynamic Compression,” Osteoarthritis Cartilage, 17(8), pp. 980–987. [CrossRef] [PubMed]
Dozin, B., Malpeli, M., Camardella, L., Cancedda, R., and Pietrangelo, A., 2002, “Response of Young, Aged and Osteoarthritic Human Articular Chondrocytes to Inflammatory Cytokines: Molecular and Cellular Aspects,” Matrix Biol., 21(5), pp. 449–459. [CrossRef] [PubMed]
Saklatvala, J., 1986, “Tumour Necrosis Factor Alpha Stimulates Resorption and Inhibits Synthesis of Proteoglycan in Cartilage,” Nature, 322(6079), pp. 547–549. [CrossRef] [PubMed]
Lefebvre, V., Peeters-Joris, C., and Vaes, G., 1990, “Modulation by Interleukin 1 and Tumor Necrosis Factor Alpha of Production of Collagenase, Tissue Inhibitor of Metalloproteinases and Collagen Types in Differentiated and Dedifferentiated Articular Chondrocytes,” Biochim. Biophys. Acta, 1052(3), pp. 366–378. [CrossRef] [PubMed]
Reginato, A. M., Sanz-Rodriguez, C., Diaz, A., Dharmavaram, R. M., and Jimenez, S. A., 1993, “Transcriptional Modulation of Cartilage-Specific Collagen Gene Expression by Interferon Gamma and Tumour Necrosis Factor Alpha in Cultured Human Chondrocytes,” Biochem. J., 294(Pt. 3), pp. 761–769. [PubMed]
Saklatvala, J., and Bird, T., 1986, “A Common Class of Receptors for the Two Types of Porcine Interleukin-1 on Articular Chondrocytes,” Lymphokine Res., 5(Suppl. 1), pp. S99–S104. [PubMed]
Campbell, I. K., Piccoli, D. S., Roberts, M. J., Muirden, K. D., and Hamilton, J. A., 1990, “Effects of Tumor Necrosis Factor Alpha and Beta on Resorption of Human Articular Cartilage and Production of Plasminogen Activator by Human Articular Chondrocytes,” Arthritis Rheum., 33(4), pp. 542–552. [CrossRef] [PubMed]
Bunning, R. A., and Russell, R. G., 1989, “The Effect of Tumor Necrosis Factor Alpha and Gamma-Interferon on the Resorption of Human Articular Cartilage and on the Production of Prostaglandin E and of Caseinase Activity by Human Articular Chondrocytes,” Arthritis Rheum., 32(6), pp. 780–784. [CrossRef] [PubMed]
Chowdhury, T. T., Bader, D. L., and Lee, D. A., 2006, “Dynamic Compression Counteracts IL-1Beta Induced iNOS and COX-2 Activity by Human Chondrocytes Cultured in Agarose Constructs,” Biorheology, 43(3–4), pp. 413–429. [PubMed]
Torzilli, P. A., Bhargava, M., Park, S., and Chen, C. T., 2010, “Mechanical Load Inhibits IL-1 Induced Matrix Degradation in Articular Cartilage,” Osteoarthritis Cartilage, 18(1), pp. 97–105. [CrossRef] [PubMed]
Patwari, P., Cook, M. N., DiMicco, M. A., Blake, S. M., James, I. E., and Kumar, S., 2003, “Proteoglycan Degradation After Injurious Compression of Bovine and Human Articular Cartilage in Vitro: Interaction With Exogenous Cytokines,” Arthritis Rheum., 48(5), pp. 1292–1301. [CrossRef] [PubMed]
Kuroki, K., Stoker, A. M., and Cook, J. L., 2005, “Effects of Proinflammatory Cytokines on Canine Articular Chondrocytes in a Three-Dimensional Culture,” Am. J. Vet. Res., 66(7), pp. 1187–1196. [CrossRef] [PubMed]
Fermor, B., Weinberg, J. B., Pisetsky, D. S., Misukonis, M. A., Banes, A. J., and Guilak, F., 2001, “The Effects of Static and Intermittent Compression on Nitric Oxide Production in Articular Cartilage Explants,” J. Orthop. Res, 19(4), pp. 729–737. [CrossRef] [PubMed]
Fermor, B., Haribabu, B., Weinberg, J. B., Pisetsky, D. S., and Guilak, F., 2001, “Mechanical Stress and Nitric Oxide Influence Leukotriene Production in Cartilage,” Biochem. Biophys. Res. Commun., 285(3), pp. 806–810. [CrossRef] [PubMed]
Bau, B., Gebhard, P. M., Haag, J., Knorr, T., Bartnik, E., and Aigner, T., 2002, “Relative Messenger RNA Expression Profiling of Collagenases and Aggrecanases in Human Articular Chondrocytes in Vivo and in Vitro,” Arthritis Rheum., 46(10), pp. 2648–2657. [CrossRef] [PubMed]
Livak, K. J., and Schmittgen, T. D., 2001, “Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2(-Delta Delta C(T)) Method,” Methods, 25(4), pp. 402–408. [CrossRef] [PubMed]
Ferretti, M., Gassner, R., Wang, Z., Perera, P., Deschner, J., Sowa, G., Salter, R. B., and Agarwal, S., 2006, “Biomechanical Signals Suppress Proinflammatory Responses in Cartilage: Early Events in Experimental Antigen-Induced Arthritis,” J. Immunol., 177, pp. 8757–8766. [CrossRef] [PubMed]
Li, Y., Frank, E. H., Wang, Y., Chubinskaya, S., Huang, H. H., and Grodzinsky, A. J., 2013, “Moderate Dynamic Compression Inhibits Pro-Catabolic Response of Cartilage to Mechanical Injury, Tumor Necrosis Factor-α and Interleukin-6, but Accentuates Degradation Above a Strain Threshold,” Osteoarthritis Cartilage, 21, pp. 1933–1941. [CrossRef] [PubMed]
Noyes, F. R., Schipplein, O. D., Andriacchi, T. P., Saddemi, S. R., and Weise, M., 1992, “The Anterior Cruciate Ligament-Deficient Knee With Varus Alignment. An Analysis of Gait Adaptations and Dynamic Joint Loadings,” Am. J. Sports Med., 20(6), pp. 707–716. [CrossRef] [PubMed]
Little, C. B., Ghosh, P., and Bellenger, C. R., 1996, “Topographic Variation in Biglycan and Decorin Synthesis by Articular Cartilage in the Early Stages of Osteoarthritis: An Experimental Study in Sheep,” J. Orthop. Res., 14(3), pp. 433–444. [CrossRef] [PubMed]
Little, C. B., and Ghosh, P., 1997, “Variation in Proteoglycan Metabolism by Articular Chondrocytes in Different Joint Regions is Determined by Post-Natal Mechanical Loading,” Osteoarthritis Cartilage, 5(1), pp. 49–62. [CrossRef] [PubMed]
Appleyard, R. C., Burkhardt, D., Ghosh, P., Read, R., Cake, M., and Swain, M. V., 2003, “Topographical Analysis of the Structural, Biochemical and Dynamic Biomechanical Properties of Cartilage in an Ovine Model of Osteoarthritis,” Osteoarthritis Cartilage, 11(1), pp. 65–77. [CrossRef] [PubMed]
Briant, P. L., 2008, “Structural Variations in Cartilage Make it Sensitive to Shifts in Joint Kinematics,” Ph.D. thesis, Stanford University, Stanford, CA.
Koo, S., Rylander, J. H., and Andriacchi, T. P., 2001, “Knee Joint Kinematics During Walking Influences the Spatial Cartilage Thickness Distribution in the Knee,” J. Biomech., 44(7), pp. 1405–1409. [CrossRef]
Li, G., Park, S. E., DeFrate, L. E., Schutzer, M. E., Ji, L., and Gill, T. J., 2005, “The Cartilage Thickness Distribution in the Tibiofemoral Joint and Its Correlation With Cartilage-to-Cartilage Contact,” Clin. Biomech. (Bristol, Avon), 20(7), pp. 736–744. [CrossRef] [PubMed]
Andriacchi, T. P., and Mundermann, A., 2006, “The Role of Ambulatory Mechanics in the Initiation and Progression of Knee Osteoarthritis,” Curr. Opin. Rheumatol., 18(5), pp. 514–518. [CrossRef] [PubMed]
Ragan, P. M., Badger, A. M., Cook, M., Chin, V. I., Gowen, M., and Grodzinsky, A. J., 1999, “Down-Regulation of Chondrocyte Aggrecan and Type-II Collagen Gene Expression Correlates With Increases in Static Compression Magnitude and Duration,” J. Orthop. Res., 17(6), pp. 836–842. [CrossRef] [PubMed]
Leipzig, N. D., and Athanasiou, K., 2008, “Static Compression of Single Chondrocytes Catabolically Modifies Single Cell Gene Expression,” J. Biophys., 94(6), pp. 2412–2422. [CrossRef]
Torzilli, P. A., Grigiene, R., Huang, C., Friedman, S. M., Doty, S. B., and Boskey, A. L., 1997, “Characterization of Cartilage Metabolic Response to Static and Dynamic Stress Using a Mechanical Explant Test System,” J. Biomech., 30(1), pp. 1–9. [CrossRef] [PubMed]
Palmoski, M. J., and Brandt, K. D., 1984, “Effects of Static and Cyclic Compressive Loading on Articular Cartilage Plugs in Vitro,” Arthritis Rheumtol., 27(6), pp. 675–681. [CrossRef]
Sah, R. L., Kim, Y. J., Doong, J. Y., Grodzinsky, A. J., Plaas, A. H., and Sandy, J. D., 1989, “Biosynthetic Response of Cartilage Explants to Dynamic Compression,” J. Orthop. Res., 7(5), pp. 619–636. [CrossRef] [PubMed]
Gray, M. L., Pizzanelli, A. M., Grodzinsky, A. J., and Lee, R. C., 1988, “Mechanical and Physiochemical Determinants of the Chondrocyte Biosynthetic Response,” J. Orthop. Res., 6(6), pp. 777–792. [CrossRef] [PubMed]
Wong, M., Siegrist, M., and Cao, X., 1999, “Cyclic Compression of Articular Cartilage Explants is Associated With Progressive Consolidation and Altered Expression Pattern of Extracellular Matrix Proteins,” Matrix Biol., 18(4), pp. 391–399. [CrossRef] [PubMed]
Li, K. W., Williamson, A. K., Wang, A. S., and Sah, R. L., 2001, “Growth Responses of Cartilage to Static and Dynamic Compression,” Clin. Orthop. Relat. Res., 391(Suppl. 1), pp. S34–S48. [CrossRef] [PubMed]
Kobayashi, M., Squires, G. R., Mousa, A., Tanzer, M., Zukor, D. J., and Antoniou, J., 2005, “Role of Interleukin-1 and Tumor Necrosis Factor Alpha in Matrix Degradation of Human Osteoarthritic Cartilage,” Arthritis. Rheum., 52(1), pp. 128–135. [CrossRef] [PubMed]
Andriacchi, T. P., 2012, “Osteoarthritis: Probing Knee OA as a System Responding to a Stimulus,” Nat. Rev. Rheumatol., 8(7), pp. 371–372. [CrossRef] [PubMed]

Figures

Grahic Jump Location
Fig. 2

Validation of the 2−ΔΔCt method. ΔCt calculated as Ct,target gene − Ct,18 s for each cDNA dilution. Data were fit using a least-squares regression analysis (with each reaction run in triplicate) to calculate the slope of the best fit line. A slope < 0.1 indicates that amplification efficiencies are adequately similar to use the 2−ΔΔCt method [31].

Grahic Jump Location
Fig. 3

Relative expression levels (averaged central and peripheral) of (a) type II collagen and aggrecan, (b) MMPs 1, 3, and 13, (c) ADAM-TS4, ADAM-TS5, and TNF-α. (d) TIMPs 1 and 2. * indicates significant difference (p < 0.05) from free-swelling control values, + indicates Load/TNF-α interaction term significant (MMP1, ADAM-TS5, TNF).

Grahic Jump Location
Fig. 1

Time course of study. Explants equilibrate in free-swelling culture for ∼48 h prior to being subjected to one of four treatment groups. After 6 h in the assigned treatment (e.g., dynamic unconfined compression), explants are digested and total RNA is isolated.

Grahic Jump Location
Fig. 4

Relative expression of ADAM-TS4, stratified by region. * indicates significant difference relative to free-swelling controls and + indicates significant regional differences (p < 0.05).

Grahic Jump Location
Fig. 5

Relative expression levels of CII (left) and aggrecan (right) stratified by region. * indicates significant difference from free-swelling controls and + indicates significant regional differences in the upregulation of mRNA (p < 0.05).

Grahic Jump Location
Fig. 6

Relative expression of MMP1 (left) and MMP3 (right) stratified by region. * indicates significant difference from free-swelling controls, + indicates significant regional differences in the upregulation of mRNA.

Tables

Errata

Discussions

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging and repositioning the boxes below.

Related Journal Articles
Related eBook Content
Topic Collections

Sorry! You do not have access to this content. For assistance or to subscribe, please contact us:

  • TELEPHONE: 1-800-843-2763 (Toll-free in the USA)
  • EMAIL: asmedigitalcollection@asme.org
Sign In