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Biomechanics and Modeling in Mechanobiology

, Volume 12, Issue 5, pp 889–899 | Cite as

Scaffold architecture determines chondrocyte response to externally applied dynamic compression

  • Tariq Mesallati
  • Conor T. Buckley
  • Thomas Nagel
  • Daniel J. Kelly
Original Paper

Abstract

It remains unclear how specific mechanical signals generated by applied dynamic compression (DC) regulate chondrocyte biosynthetic activity. It has previously been suggested that DC-induced interstitial fluid flow positively impacts cartilage-specific matrix production. Modifying fluid flow within dynamically compressed hydrogels therefore represents a promising approach to controlling chondrocyte behavior, which could potentially be achieved by changing the construct architecture. The objective of this study was to first determine the influence of construct architecture on the mechanical environment within dynamically compressed agarose hydrogels using finite element (FE) modeling and to then investigate how chondrocytes would respond to this altered environment. To modify construct architecture, an array of channels was introduced into the hydrogels. Increased magnitudes of fluid flow were predicted in the periphery of dynamically compressed solid hydrogels and also around the channels in the dynamically compressed channeled hydrogels. DC was found to significantly increase sGAG synthesis in solid constructs, which could be attributed at least in part to an increase in DNA. DC was also found to preferentially increase collagen accumulation in regions of solid and channeled constructs where FE modeling predicted higher levels of fluid flow, suggesting that this stimulus is important for promoting collagen production by chondrocytes embedded in agarose gels. In conclusion, this study demonstrates how the architecture of cell-seeded scaffolds or hydrogels can be modified to alter the spatial levels of biophysical cues throughout the construct, leading to greater collagen accumulation throughout the engineered tissue rather than preferentially in the construct periphery. This system also provides a novel approach to investigate how chondrocytes respond to altered levels of biophysical stimulation.

Keywords

Dynamic compression Agarose hydrogel Chondrocytes Channeled constructs Biophysical stimulation Nutrient transport 

Abbreviations

DC

Dynamic compression

FE

Finite element

sGAG

Sulfated glycosaminoglycan

ECM

Extracellular matrix

PBS

Phosphate-buffered saline

hgDMEM

High-glucose Dulbecco’s modified Eagle’s medium

FBS

Fetal bovine serum

DMSO

Dimethyl sulphoxide

P1

Passage one

FGF-2

Fibroblast growth factor-2

CDM

Chondrogenic medium

PTFE

Polytetrafluoroethylene

PDMS

Polydimethylsiloxane

CNC

Computer numerical controlled

TGF-β 3

Transforming growth factor-beta 3

FS

Free swelling

DCS

Dynamically compressed solid

DCM

Dynamically compressed microchannel

FSM

Free swelling microchannel

FSS

Free swelling solid

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Supplementary material

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References

  1. Albro MB, Li R, Banerjee RE, Hung CT, Ateshian GA (2010) Validation of theoretical framework explaining active solute uptake in dynamically loaded porous media. J Biomech 43(12): 2267–2273CrossRefGoogle Scholar
  2. Appelman TP, Mizrahi J, Elisseeff JH, Seliktar D (2009) The differential effect of scaffold composition and architecture on chondrocyte response to mechanical stimulation. Biomaterials 30(4): 518–525CrossRefGoogle Scholar
  3. Appelman TP, Mizrahi J, Elisseeff JH, Seliktar D (2011) The influence of biological motifs and dynamic mechanical stimulation in hydrogel scaffold systems on the phenotype of chondrocytes. Biomaterials 32(6): 1508–1516CrossRefGoogle Scholar
  4. Benya PD, Shaffer JD (1982) Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 30(1): 215–224CrossRefGoogle Scholar
  5. Bian L, Angione SL, Ng KW, Lima EG, Williams DY, Mao DQ, Ateshian GA, Hung CT (2009) Influence of decreasing nutrient path length on the development of engineered cartilage. Osteoarthr Cartil 17(5): 677–685CrossRefGoogle Scholar
  6. Bryant SJ, Chowdhury TT, Lee DA, Bader DL, Anseth KS (2004) Crosslinking density influences chondrocyte metabolism in dynamically loaded photocrosslinked poly(ethylene glycol) hydrogels. Ann Biomed Eng 32(3): 407–417CrossRefGoogle Scholar
  7. Bryant SJ, Nicodemus GD, Villanueva I (2008) Designing 3D photopolymer hydrogels to regulate biomechanical cues and tissue growth for cartilage tissue engineering. Pharm Res 25(10): 2379–2386CrossRefGoogle Scholar
  8. Buckley CT, Thorpe SD, Kelly DJ (2009a) Engineering of large cartilaginous tissues through the use of microchanneled hydrogels and rotational culture. Tissue Eng Part A 15(11): 3213–3220CrossRefGoogle Scholar
  9. Buckley CT, Thorpe SD, O’Brien FJ, Robinson AJ, Kelly DJ (2009b) The effect of concentration, thermal history and cell seeding density on the initial mechanical properties of agarose hydrogels. J Mech Behav Biomed Mater 2(5): 512–521CrossRefGoogle Scholar
  10. Buschmann MD, Gluzband YA, Grodzinsky AJ, Kimura JH, Hunziker EB (1992) Chondrocytes in agarose culture synthesize a mechanically functional extracellular matrix. J Orthop Res 10(6): 745–758. doi: 10.1002/jor.1100100602 CrossRefGoogle Scholar
  11. Buschmann MD, Gluzband YA, Grodzinsky AJ, Hunziker EB (1995) Mechanical compression modulates matrix biosynthesis in chondrocyte/agarose culture. J Cell Sci 108(Pt 4): 1497–1508Google Scholar
  12. Buschmann MD, Kim YJ, Wong M, Frank E, Hunziker EB, Grodzinsky AJ (1999) Stimulation of aggrecan synthesis in cartilage explants by cyclic loading is localized to regions of high interstitial fluid flow. Arch Biochem Biophys 366(1): 1–7CrossRefGoogle Scholar
  13. Chowdhury TT, Bader DL, Lee DA (2001) Dynamic compression inhibits the synthesis of nitric oxide and PGE2 by IL-1β-stimulated chondrocytes cultured in agarose constructs. Biochem Biophys Res Commun 285(5): 1168–1174CrossRefGoogle Scholar
  14. Cowin SC (2002) Mechanosensation and fluid transport in living bone. J Musculoskelet Neuronal Interact 2(3): 256–260Google Scholar
  15. Cowin SC (2007) The significance of bone microstructure in mechanotransduction. J Biomech 40(Suppl. 1): S105–S109MathSciNetCrossRefGoogle Scholar
  16. Gemmiti CV, Guldberg RE (2006) Fluid flow increases type II collagen deposition and tensile mechanical properties in bioreactor-grown tissue-engineered cartilage. Tissue Eng 12(3): 469–479CrossRefGoogle Scholar
  17. Gu WY, Yao H, Huang CY, Cheung HS (2003) New insight into deformation-dependent hydraulic permeability of gels and cartilage, and dynamic behavior of agarose gels in confined compression. J Biomech 36(4): 593–598CrossRefGoogle Scholar
  18. Grodzinsky AJ, Levenston ME, Jin M, Frank EH (2000) Cartilage tissue remodeling in response to mechanical forces. Annu Rev Biomed Eng 2: 691–713. doi: 10.1146/annurev.bioeng.2.1.691 CrossRefGoogle Scholar
  19. Hunter C, Mouw JK, Levenston ME (2004) Dynamic compression of chondrocyte-seeded fibrin gels: effects on matrix accumulation and mechanical stiffness. Osteoarthr Cartil 12(2): 117–130CrossRefGoogle Scholar
  20. Ignat’eva NY, Danilov NA, Averkiev SV, Obrezkova MV, Lunin VV, Sobol EN (2007) Determination of hydroxyproline in tissues and the evaluation of the collagen content of the tissues. J Ann Chem 62(1): 51–57CrossRefGoogle Scholar
  21. Kafienah W, Sims TJ (2004) Biochemical methods for the analysis of tissue-engineered cartilage. Methods Mol Biol 238: 217–230Google Scholar
  22. Kelly DJ, Prendergast PJ (2005) Mechano-regulation of stem cell differentiation and tissue regeneration in osteochondral defects. J Biomech 38(7): 1413–1422CrossRefGoogle Scholar
  23. Kelly T-AN, Wang CCB, Mauck RL, Ateshian GA, Hung CT (2004) Role of cell-associated matrix in the development of free-swelling and dynamically loaded chondrocyte-seeded agarose gels. Biorheology 41(3–4): 223–237Google Scholar
  24. Kelly T-AN, Ng KW, Ateshian GA, Hung CT (2009) Analysis of radial variations in material properties and matrix composition of chondrocyte-seeded agarose hydrogel constructs. Osteoarthr Cartil 17(1): 73–82CrossRefGoogle Scholar
  25. Kim YJ, Sah RL, Doong JY, Grodzinsky AJ (1988) Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem 174(1): 168–176CrossRefGoogle Scholar
  26. Kim YJ, Sah RLY, Grodzinsky AJ, Plaas AHK, Sandy JD (1994) Mechanical regulation of cartilage biosynthetic behavior: physical stimuli. Arch Biochem Biophys 311(1): 1–12CrossRefGoogle Scholar
  27. Lee DA, Bader DL (1997) Compressive strains at physiological frequencies influence the metabolism of chondrocytes seeded in agarose. J Orthop Res 15(2): 181–188CrossRefGoogle Scholar
  28. Lee DA, Frean SP, Lees P, Bader DL (1998) Dynamic mechanical compression influences nitric oxide production by articular chondrocytes seeded in agarose. Biochem Biophys Res Commun 251(2): 580–585CrossRefGoogle Scholar
  29. Lee DA, Noguchi T, Frean SP, Lees P, Bader DL (2000) The influence of mechanical loading on isolated chondrocytes seeded in agarose constructs. Biorheology 37(1–2): 149–161Google Scholar
  30. MSC (2008) Marc 2008r1–volume A: theory and user information. MSC.Software Corporation, 2 MacArthur Place, Santa Ana, CA 92707, USA. http://www.mscsoftware.com
  31. Martin I, Obradovic B, Treppo S, Grodzinsky AJ, Langer R, Freed LE, Vunjak-Novakovic G (2000) Modulation of the mechanical properties of tissue engineered cartilage. Biorheology 37(1–2): 141–147Google Scholar
  32. Mauck RL, Soltz MA, Wang CC, Wong DD, Chao PH, Valhmu WB, Hung CT, Ateshian GA (2000) Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. J Biomech Eng 122(3): 252–260CrossRefGoogle Scholar
  33. Mauck RL, Seyhan SL, Ateshian GA, Hung CT (2002) Influence of seeding density and dynamic deformational loading on the developing structure/function relationships of chondrocyte-seeded agarose hydrogels. Ann Biomed Eng 30(8): 1046–1056CrossRefGoogle Scholar
  34. Mauck RL, Hung CT, Ateshian GA (2003a) Modeling of neutral solute transport in a dynamically loaded porous permeable gel: implications for articular cartilage biosynthesis and tissue engineering. J Biomech Eng 125(5): 602–614CrossRefGoogle Scholar
  35. Mauck RL, Nicoll SB, Seyhan SL, Ateshian GA, Hung CT (2003b) Synergistic action of growth factors and dynamic loading for articular cartilage tissue engineering. Tissue Eng 9(4): 597–611. doi: 10.1089/107632703768247304 CrossRefGoogle Scholar
  36. Mauck RL, Byers BA, Yuan X, Tuan RS (2007) Regulation of cartilaginous ECM gene transcription by chondrocytes and MSCs in 3D culture in response to dynamic loading. Biomech Model Mechanobiol 6(1–2): 113–125CrossRefGoogle Scholar
  37. Ng KW, Mauck RL, Statman LY, Lin EY, Ateshian GA, Hung CT (2006) Dynamic deformational loading results in selective application of mechanical stimulation in a layered, tissue-engineered cartilage construct. Biorheology 43(3–4): 497–507Google Scholar
  38. Prendergast PJ, Huiskes R, Søballe K (1997) Biophysical stimuli on cells during tissue differentiation at implant interfaces. J Biomech 30(6): 539–548CrossRefGoogle Scholar
  39. Sheehy EJ, Buckley CT, Kelly DJ (2011) Chondrocytes and bone marrow-derived mesenchymal stem cells undergoing chondrogenesis in agarose hydrogels of solid and channelled architectures respond differentially to dynamic culture conditions. J Tissue Eng Regen Med 5(9): 747–758CrossRefGoogle Scholar
  40. Silva MMCG, Cyster LA, Barry JJA, Yang XB, Oreffo ROC, Grant DM, Scotchford CA, Howdle SM, Shakesheff KM, Rose FRAJ (2006) The effect of anisotropic architecture on cell and tissue infiltration into tissue engineering scaffolds. Biomaterials 27(35): 5909–5917CrossRefGoogle Scholar
  41. Sun D, Aydelotte MB, Maldonado B (1986) Clonal analysis of the population of chondrocytes from the Swarm rat chondrosarcoma in agarose culture. J Orthop Res 4(4): 427–436CrossRefGoogle Scholar
  42. Thorpe SD, Buckley CT, Vinardell T, O’Brien FJ, Campbell VA, Kelly DJ (2010) The response of bone marrow-derived mesenchymal stem cells to dynamic compression following tgf-β3 induced chondrogenic differentiation. Ann Biomed Eng 38(9):2896–2909Google Scholar
  43. Villanueva I, Hauschulz DS, Mejic D, Bryant SJ (2008) Static and dynamic compressive strains influence nitric oxide production and chondrocyte bioactivity when encapsulated in PEG hydrogels of different crosslinking densities. Osteoarthr Cartil 16(8): 909–918CrossRefGoogle Scholar
  44. Villanueva I, Gladem SK, Kessler J, Bryant SJ (2010) Dynamic loading stimulates chondrocyte biosynthesis when encapsulated in charged hydrogels prepared from poly(ethylene glycol) and chondroitin sulfate. Matrix Biol 29(1): 51–62CrossRefGoogle Scholar
  45. Waldman SD, Couto DC, Grynpas MD, Pilliar RM, Kandel RA (2006) A single application of cyclic loading can accelerate matrix deposition and enhance the properties of tissue-engineered cartilage. Osteoarthr Cartil 14(4): 323–330CrossRefGoogle Scholar
  46. Wang QG, Magnay JL, Nguyen B, Thomas CR, Zhang Z, El Haj AJ, Kuiper NJ (2009) Gene expression profiles of dynamically compressed single chondrocytes and chondrons. Biochem Biophys Res Commun 379(3): 738–742CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Tariq Mesallati
    • 1
    • 2
  • Conor T. Buckley
    • 1
    • 2
  • Thomas Nagel
    • 1
    • 2
    • 3
  • Daniel J. Kelly
    • 1
    • 2
  1. 1.Department of Mechanical and Manufacturing Engineering, School of EngineeringTrinity College DublinDublinIreland
  2. 2.Trinity Centre for Bioengineering, Trinity Biomedical Sciences InstituteTrinity College DublinDublinIreland
  3. 3.Department of Environmental InformaticsHelmholtz Centre for Environmental Research-UFZLeipzigGermany

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