The Effect of Intermittent Static Biaxial Tensile Strains on Tissue Engineered Cartilage

Mechanical stimulation of engineered cartilage constructs is a commonly applied method used to accelerate tissue formation and improve the mechanical properties of the developed tissue. While the effects of compression and shear have been widely studied, the effect of tension has received relatively little attention. As articular cartilage in vivo is subjected to a degree of static tension (pre-tension) even in the absence of externally applied loads, the purpose of this study was to investigate the effect of intermittent static biaxial tensile strains (BTS) on chondrocyte metabolism and resultant tissue formation. Using a custom-design loading fixture to apply BTS, the optimal conditions for stimulating extracellular matrix synthesis were under average magnitudes of 3.8% radial and 2.1% circumferential tensile strains for 30 min. Tissue constructs subjected to tensile strain stimulation 3 times/week for a period of 4 weeks displayed increased thickness (35 ± 18%) and proteoglycan content (22 ± 7%) without an associated change in mechanical properties. In contrast, constructs stimulated daily over the same time period exhibited negligible effects in terms of ECM accumulation suggesting that the frequency of stimulation needs to be precisely controlled. The results of this study demonstrate that while tension can be used as potential biomechanical stimulus to improve tissue formation, further optimization of this process needs to be conducted to improve ECM accumulation and tissue mechanical properties after long-term exposure to tensile stimuli.

[1]  R. Diegelmann,et al.  Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. , 1971, Biochemistry.

[2]  Stuart J Warden,et al.  Cellular accommodation and the response of bone to mechanical loading. , 2005, Journal of biomechanics.

[3]  M. Buschmann,et al.  Optimization of Histoprocessing Methods to Detect Glycosaminoglycan, Collagen Type II, and Collagen Type I in Decalcified Rabbit Osteochondral Sections , 2005 .

[4]  E B Hunziker,et al.  Mechanical compression alters proteoglycan deposition and matrix deformation around individual cells in cartilage explants. , 1998, Journal of cell science.

[5]  Sheldon R. Simon,et al.  Orthopaedic basic science : biology and biomechanics of the musculoskeletal system , 2000 .

[6]  J. F. Woessner,et al.  The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. , 1961, Archives of biochemistry and biophysics.

[7]  E B Hunziker,et al.  Mechanical compression modulates matrix biosynthesis in chondrocyte/agarose culture. , 1995, Journal of cell science.

[8]  S. Waldman,et al.  Effect of Biomechanical Conditioning on Cartilaginous Tissue Formation in Vitro , 2003, The Journal of bone and joint surgery. American volume.

[9]  S. Waldman,et al.  Mechanical vibrations increase the proliferation of articular chondrocytes in high-density culture , 2008, Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine.

[10]  N. Ramakrishnan,et al.  Effective elastic moduli of porous solids , 1990 .

[11]  L. Ballou,et al.  Cyclic equibiaxial tensile strain induces both anabolic and catabolic responses in articular chondrocytes. , 2007, Gene.

[12]  R Langer,et al.  Modulation of the mechanical properties of tissue engineered cartilage. , 2000, Biorheology.

[13]  H. Fry The interlocked stresses of articular cartilage. , 1974, British Journal of Plastic Surgery.

[14]  E B Hunziker,et al.  Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. , 2002, Osteoarthritis and cartilage.

[15]  C. P. Leblond,et al.  Thymidine-H3 as a tool for the investigation of the renewal of cell populations. , 1959, Laboratory investigation; a journal of technical methods and pathology.

[16]  A. Grodzinsky,et al.  Fluorometric assay of DNA in cartilage explants using Hoechst 33258. , 1988, Analytical biochemistry.

[17]  Andrés J. García,et al.  Oscillatory tension differentially modulates matrix metabolism and cytoskeletal organization in chondrocytes and fibrochondrocytes. , 2004, Journal of biomechanics.

[18]  K. Naruse,et al.  Effects of tensile and compressive strains on response of a chondrocytic cell line embedded in type I collagen gel. , 2008, Journal of biotechnology.

[19]  Moonsoo Jin,et al.  Effects of dynamic compressive loading on chondrocyte biosynthesis in self-assembling peptide scaffolds. , 2004, Journal of biomechanics.

[20]  M. Wong,et al.  Cyclic tensile strain and cyclic hydrostatic pressure differentially regulate expression of hypertrophic markers in primary chondrocytes. , 2003, Bone.

[21]  A. Grodzinsky,et al.  Chondrocytes in agarose culture synthesize a mechanically functional extracellular matrix , 1992, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[22]  S. Waldman,et al.  The effect of continuous culture on the growth and structure of tissue‐engineered cartilage , 2009, Biotechnology progress.

[23]  D. Narmoneva,et al.  Nonuniform swelling-induced residual strains in articular cartilage. , 1999, Journal of biomechanics.

[24]  D. Heinegård,et al.  Biochemistry and Metabolism of Normal and Osteoarthritic Cartilage , 2003 .

[25]  W M Lai,et al.  A triphasic theory for the swelling and deformation behaviors of articular cartilage. , 1991, Journal of biomechanical engineering.

[26]  R. Kandel,et al.  Long‐term intermittent shear deformation improves the quality of cartilaginous tissue formed in vitro , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[27]  W. Davis,et al.  The distortion of autogenous cartilage grafts: Its cause and prevention , 1957 .

[28]  S. Waldman,et al.  Long-term intermittent compressive stimulation improves the composition and mechanical properties of tissue-engineered cartilage. , 2004, Tissue engineering.

[29]  L. Bonassar,et al.  The effect of dynamic compression on the response of articular cartilage to insulin‐like growth factor‐I , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[30]  R. L. Goldberg,et al.  An improved method for determining proteoglycans synthesized by chondrocytes in culture. , 1990, Connective tissue research.

[31]  T. Koob,et al.  Early changes in material properties of rabbit articular cartilage after meniscectomy , 1983, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[32]  D L Bader,et al.  The influence of mechanical loading on isolated chondrocytes seeded in agarose constructs. , 2000, Biorheology.

[33]  D. Howard,et al.  Tissue engineering strategies for cartilage generation--micromass and three dimensional cultures using human chondrocytes and a continuous cell line. , 2005, Biochemical and biophysical research communications.

[34]  R. Kandel,et al.  Cyclic compressive mechanical stimulation induces sequential catabolic and anabolic gene changes in chondrocytes resulting in increased extracellular matrix accumulation. , 2006, Matrix biology : journal of the International Society for Matrix Biology.

[35]  B. Obradovic,et al.  Bioreactor cultivation conditions modulate the composition and mechanical properties of tissue‐engineered cartilage , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[36]  A. Hall,et al.  Control of matrix synthesis in isolated bovine chondrocytes by extracellular and intracellular pH , 1995, Journal of cellular physiology.

[37]  R. Spencer,et al.  Hyaline cartilage engineered by chondrocytes in pellet culture: histological, immunohistochemical and ultrastructural analysis in comparison with cartilage explants , 2004, Journal of anatomy.

[38]  M E Levenston,et al.  Articular chondrocytes derived from distinct tissue zones differentially respond to in vitro oscillatory tensile loading. , 2008, Osteoarthritis and cartilage.

[39]  Y. Kato,et al.  The effects of high magnitude cyclic tensile load on cartilage matrix metabolism in cultured chondrocytes. , 2000, European journal of cell biology.

[40]  A. Maroudas,et al.  Chemical composition and swelling of normal and osteoarthrotic femoral head cartilage. II. Swelling. , 1977, Annals of the rheumatic diseases.

[41]  A. Grodzinsky,et al.  Tissue shear deformation stimulates proteoglycan and protein biosynthesis in bovine cartilage explants. , 2001, Archives of biochemistry and biophysics.

[42]  S. Waldman,et al.  Effect of sodium bicarbonate on extracellular pH, matrix accumulation, and morphology of cultured articular chondrocytes. , 2004, Tissue engineering.

[43]  R. Kandel,et al.  Characterization of proteoglycan accumulation during formation of cartilagenous tissue in vitro. , 1995, Osteoarthritis and cartilage.

[44]  B. Månsson,et al.  On the enzymatic exchange of the sulfate group of chondroitinsulfuric acid in slices of cartilage. , 1952, The Journal of biological chemistry.

[45]  C H Turner,et al.  Three rules for bone adaptation to mechanical stimuli. , 1998, Bone.

[46]  J. Connelly,et al.  The influence of cyclic tension amplitude on chondrocyte matrix synthesis: experimental and finite element analyses. , 2004, Biorheology.

[47]  D. Schurman,et al.  Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[48]  W. Hayes,et al.  A mathematical analysis for indentation tests of articular cartilage. , 1972, Journal of biomechanics.

[49]  S. Waldman,et al.  Characterization of cartilagenous tissue formed on calcium polyphosphate substrates in vitro. , 2002, Journal of biomedical materials research.

[50]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[51]  A. Robling,et al.  Mechanosensitivity of the rat skeleton decreases after a long period of loading, but is improved with time off. , 2005, Bone.

[52]  G A Ateshian,et al.  Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. , 2000, Journal of biomechanical engineering.