The effects of dynamic loading on the intervertebral disc

Loading is important to maintain the balance of matrix turnover in the intervertebral disc (IVD). Daily cyclic diurnal assists in the transport of large soluble factors across the IVD and its surrounding circulation and applies direct and indirect stimulus to disc cells. Acute mechanical injury and accumulated overloading, however, could induce disc degeneration. Recently, there is more information available on how cyclic loading, especially axial compression and hydrostatic pressure, affects IVD cell biology. This review summarises recent studies on the response of the IVD and stem cells to applied cyclic compression and hydrostatic pressure. These studies investigate the possible role of loading in the initiation and progression of disc degeneration as well as quantifying a physiological loading condition for the study of disc degeneration biological therapy. Subsequently, a possible physiological/beneficial loading range is proposed. This physiological/beneficial loading could provide insight into how to design loading regimes in specific system for the testing of various biological therapies such as cell therapy, chemical therapy or tissue engineering constructs to achieve a better final outcome. In addition, the parameter space of ‘physiological’ loading may also be an important factor for the differentiation of stem cells towards most ideally ‘discogenic’ cells for tissue engineering purpose.

[1]  L. Setton,et al.  Cell Mechanics and Mechanobiology in the Intervertebral Disc , 2004, Spine.

[2]  S Holm,et al.  Nutrition of the intervertebral disc: effect of fluid flow on solute transport. , 1982, Clinical orthopaedics and related research.

[3]  A. Ignatius,et al.  Influence of low glucose supply on the regulation of gene expression by nucleus pulposus cells and their responsiveness to mechanical loading. , 2010, Journal of neurosurgery. Spine.

[4]  D S McNally,et al.  Effects of hydrostatic pressure on matrix synthesis in different regions of the intervertebral disk. , 1996, Journal of applied physiology.

[5]  D. Haschtmann,et al.  Influence of diurnal hyperosmotic loading on the metabolism and matrix gene expression of a whole‐organ intervertebral disc model , 2006, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[6]  A. Hall,et al.  Volume‐sensitive taurine transport in bovine articular chondrocytes. , 1995, The Journal of physiology.

[7]  W. Gu,et al.  Effects of mechanical compression on metabolism and distribution of oxygen and lactate in intervertebral disc. , 2008, Journal of biomechanics.

[8]  A. Shirazi-Adl,et al.  Investigation of solute concentrations in a 3D model of intervertebral disc , 2009, European Spine Journal.

[9]  A. Freemont,et al.  Intervertebral Disc Cell–Mediated Mesenchymal Stem Cell Differentiation , 2006, Stem cells.

[10]  S. Goldstein,et al.  Effect of compressive loading on chondrocyte differentiation in agarose cultures of chick limb‐bud cells , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[11]  G. Sukhikh,et al.  Mesenchymal Stem Cells , 2002, Bulletin of Experimental Biology and Medicine.

[12]  Pascal Richette,et al.  Cyclic tensile stretch modulates proteoglycan production by intervertebral disc annulus fibrosus cells through production of nitrite oxide , 2003, Journal of cellular biochemistry.

[13]  G. Murrell,et al.  The Effect of Running Exercise on Intervertebral Disc Extracellular Matrix Production in a Rat Model , 2010, Spine.

[14]  K. Cheung,et al.  Cryopreserved intervertebral disc with injected bone marrow-derived stromal cells: a feasibility study using organ culture. , 2010, The spine journal : official journal of the North American Spine Society.

[15]  J. Urban,et al.  The role of the physicochemical environment in determining disc cell behaviour. , 2002, Biochemical Society transactions.

[16]  Shuichi Mizuno,et al.  The effect of hydrostatic pressure on three-dimensional chondroinduction of human adipose-derived stem cells. , 2009, Tissue engineering. Part A.

[17]  Farshid Guilak,et al.  The micromechanical environment of intervertebral disc cells determined by a finite deformation, anisotropic, and biphasic finite element model. , 2003, Journal of biomechanical engineering.

[18]  S. Thorpe,et al.  Dynamic compression can inhibit chondrogenesis of mesenchymal stem cells. , 2008, Biochemical and biophysical research communications.

[19]  Keita Ito,et al.  Fluid flow and convective transport of solutes within the intervertebral disc. , 2004, Journal of biomechanics.

[20]  L. Setton,et al.  Osmolarity Regulates Gene Expression in Intervertebral Disc Cells Determined by Gene Array and Real-Time Quantitative RT-PCR , 2005, Annals of Biomedical Engineering.

[21]  W C Hutton,et al.  The effect of hydrostatic pressure on intervertebral disc metabolism. , 1999, Spine.

[22]  K. Jepsen,et al.  Cyclic hydrostatic pressure enhances the chondrogenic phenotype of human mesenchymal progenitor cells differentiated in vitro , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[23]  L. Setton,et al.  Mechanobiology of the intervertebral disc and relevance to disc degeneration. , 2006, The Journal of bone and joint surgery. American volume.

[24]  Keita Ito,et al.  Effect of TGF beta1, BMP-2 and hydraulic pressure on chondrogenic differentiation of bovine bone marrow mesenchymal stromal cells. , 2009, Biorheology.

[25]  A R Hargens,et al.  Intervertebral disc nutrition. Diffusion versus convection. , 1986, Clinical orthopaedics and related research.

[26]  H. Tsuji,et al.  Water Diffusion Pathway, Swelling Pressure, and Biomechanical Properties of the Intervertebral Disc During Compression Load , 1989, Spine.

[27]  David A Lee,et al.  Dynamic compressive strain influences chondrogenic gene expression in human mesenchymal stem cells. , 2006, Biorheology.

[28]  L. Dai,et al.  Biologic Response of the Intervertebral Disc to Static and Dynamic Compression In Vitro , 2007, Spine.

[29]  P J Prendergast,et al.  Biophysical stimuli on cells during tissue differentiation at implant interfaces , 1997 .

[30]  I. Shapiro,et al.  Osmolarity and Intracellular Calcium Regulate Aquaporin2 Expression Through TonEBP in Nucleus Pulposus Cells of the Intervertebral Disc , 2009, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[31]  T. Andriacchi,et al.  Chondrocyte cells respond mechanically to compressive loads , 1994, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[32]  T. Albert,et al.  Differentiation of Mesenchymal Stem Cells Towards a Nucleus Pulposus-like Phenotype In Vitro: Implications for Cell-Based Transplantation Therapy , 2004, Spine.

[33]  A. Race,et al.  Effect of loading rate and hydration on the mechanical properties of the disc. , 2000, Spine.

[34]  Keita Ito,et al.  An In Vitro Organ Culturing System for Intervertebral Disc Explants With Vertebral Endplates: A Feasibility Study With Ovine Caudal Discs , 2006, Spine.

[35]  Daniel H K Chow,et al.  Changes in nuclear composition following cyclic compression of the intervertebral disc in an in vivo rat-tail model. , 2004, Medical engineering & physics.

[36]  Mehran Kasra,et al.  Effect of dynamic hydrostatic pressure on rabbit intervertebral disc cells , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[37]  Jeffrey C Lotz,et al.  Biological response of the intervertebral disc to dynamic loading. , 2004, Journal of biomechanics.

[38]  Luo Zj,et al.  Light and low-frequency pulsatile hydrostatic pressure enhances extracellular matrix formation by bone marrow mesenchymal cells: an in-vitro study with special reference to cartilage repair. , 2007 .

[39]  M. Doran,et al.  Enhanced Chondrogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells in Low Oxygen Environment Micropellet Cultures , 2010, Cell transplantation.

[40]  Elizabeth G Loboa,et al.  Differential effects on messenger ribonucleic acid expression by bone marrow-derived human mesenchymal stem cells seeded in agarose constructs due to ramped and steady applications of cyclic hydrostatic pressure. , 2007, Tissue engineering.

[41]  Hai Yao,et al.  Physical signals and solute transport in human intervertebral disc during compressive stress relaxation: 3D finite element analysis. , 2006, Biorheology.

[42]  Mauro Alini,et al.  Anabolic and catabolic mRNA levels of the intervertebral disc vary with the magnitude and frequency of in vivo dynamic compression , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[43]  H. Cheung,et al.  Temporal Expression Patterns and Corresponding Protein Inductions of Early Responsive Genes in Rabbit Bone Marrow–Derived Mesenchymal Stem Cells Under Cyclic Compressive Loading , 2005, Stem cells.

[44]  Keita Ito,et al.  2004 Young Investigator Award Winner: Vertebral Endplate Marrow Contact Channel Occlusions and Intervertebral Disc Degeneration , 2005, Spine.

[45]  S H Elder,et al.  Cyclic hydrostatic compression stimulates chondroinduction of C3H/10T1/2 cells , 2005, Biomechanics and modeling in mechanobiology.

[46]  Keita Ito,et al.  Effects of Immobilization and Dynamic Compression on Intervertebral Disc Cell Gene Expression In Vivo , 2003, Spine.

[47]  L. Claes,et al.  Regulation of gene expression in intervertebral disc cells by low and high hydrostatic pressure , 2006, European Spine Journal.

[48]  H. Tsuji,et al.  Effects of Hydrostatic Pressure on Matrix Synthesis and Matrix Metalloproteinase Production in the Human Lumbar Intervertebral Disc , 1997, Spine.

[49]  C. Bowman,et al.  Mechanotransducing ion channels in C6 glioma cells , 1996, Glia.

[50]  G. Beaupré,et al.  Hydrostatic Pressure Enhances Chondrogenic Differentiation of Human Bone Marrow Stromal Cells in Osteochondrogenic Medium , 2008, Annals of Biomedical Engineering.

[51]  A. Freemont,et al.  Altered integrin mechanotransduction in human nucleus pulposus cells derived from degenerated discs. , 2009, Arthritis and rheumatism.

[52]  Young Ha Kim,et al.  The effects of dynamic and three-dimensional environments on chondrogenic differentiation of bone marrow stromal cells , 2009, Biomedical materials.

[53]  S. Nicoll,et al.  Hydrostatic Pressure Differentially Regulates Outer and Inner Annulus Fibrosus Cell Matrix Production in 3D Scaffolds , 2008, Annals of Biomedical Engineering.

[54]  H Weinans,et al.  Cell and nucleus deformation in compressed chondrocyte-alginate constructs: temporal changes and calculation of cell modulus. , 2002, Biochimica et biophysica acta.

[55]  A. Maroudas,et al.  Diurnal fluid expression and activity of intervertebral disc cells. , 2006, Biorheology.

[56]  D. Sakai,et al.  Differential Phenotype of Intervertebral Disc Cells: Microarray and Immunohistochemical Analysis of Canine Nucleus Pulposus and Anulus Fibrosus , 2009, Spine.

[57]  Lorenzo Moroni,et al.  Differential Response of Adult and Embryonic Mesenchymal Progenitor Cells to Mechanical Compression in Hydrogels , 2007, Stem cells.

[58]  A. Freemont,et al.  Human cells derived from degenerate intervertebral discs respond differently to those derived from non-degenerate intervertebral discs following application of dynamic hydrostatic pressure. , 2008, Biorheology.

[59]  P M Bongers,et al.  Back pain and exposure to whole body vibration in helicopter pilots. , 1990, Ergonomics.

[60]  W. Hutton,et al.  Diurnal changes in spinal mechanics and their clinical significance. , 1990, The Journal of bone and joint surgery. British volume.

[61]  J. Taboas,et al.  Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype , 2010, Stem Cell Research & Therapy.

[62]  W. Bloch,et al.  Human mesenchymal stem cells: Influence of oxygen pressure on proliferation and chondrogenic differentiation in fibrin glue in vitro. , 2009, Journal of biomedical materials research. Part A.

[63]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[64]  David G. Wilder,et al.  Low Back Pain and Whole Body Vibration , 1998 .

[65]  J. Connelly,et al.  Dynamic Compression Regulates the Expression and Synthesis of Chondrocyte‐Specific Matrix Molecules in Bone Marrow Stromal Cells , 2007, Stem cells.

[66]  Dong Hwa Kim,et al.  Enhanced differentiation of mesenchymal stem cells into NP-like cells via 3D co-culturing with mechanical stimulation. , 2009, Journal of bioscience and bioengineering.

[67]  G Garbutt,et al.  Effect of sustained loading on the water content of intervertebral discs: implications for disc metabolism. , 1996, Annals of the rheumatic diseases.

[68]  J. Iatridis,et al.  Dynamic Compression Effects on Intervertebral Disc Mechanics and Biology , 2008, Spine.

[69]  A. Freemont,et al.  Open Access Research Article Transcriptional Profiling of Bovine Intervertebral Disc Cells: Implications for Identification of Normal and Degenerate Human Intervertebral Disc Cell Phenotypes , 2022 .

[70]  L. Setton,et al.  Matrix protein gene expression in intervertebral disc cells subjected to altered osmolarity. , 2002, Biochemical and biophysical research communications.

[71]  Steven A. Goldstein,et al.  Chondrocyte Differentiation is Modulated by Frequency and Duration of Cyclic Compressive Loading , 2001, Annals of Biomedical Engineering.

[72]  W C Hutton,et al.  Do the intervertebral disc cells respond to different levels of hydrostatic pressure? , 2001, Clinical biomechanics.

[73]  Leena Peltonen,et al.  The Twin Spine Study: contributions to a changing view of disc degeneration. , 2009, The spine journal : official journal of the North American Spine Society.

[74]  Zhen Li,et al.  Chondrogenesis of human bone marrow mesenchymal stem cells in fibrin-polyurethane composites. , 2009, Tissue engineering. Part A.

[75]  Ian A. F. Stokes,et al.  Mechanical Conditions That Accelerate Intervertebral Disc Degeneration: Overload Versus Immobilization , 2004, Spine.

[76]  C. V. van Donkelaar,et al.  Inhibition of vertebral endplate perfusion results in decreased intervertebral disc intranuclear diffusive transport , 2007, Journal of anatomy.

[77]  K. Wenger,et al.  Matrix Remodeling Expression in Anulus Cells Subjected to Increased Compressive Load , 2005, Spine.

[78]  V. Mow,et al.  Chondrocyte deformation and local tissue strain in articular cartilage: A confocal microscopy study , 1995, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[79]  N Bogduk,et al.  The pathophysiology of disc degeneration: a critical review. , 2008, The Journal of bone and joint surgery. British volume.

[80]  Stuart B Goodman,et al.  Effects of hydrostatic pressure and transforming growth factor-beta 3 on adult human mesenchymal stem cell chondrogenesis in vitro. , 2006, Tissue engineering.

[81]  L. Claes,et al.  Influence of extracellular osmolarity and mechanical stimulation on gene expression of intervertebral disc cells , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[82]  S. Milz,et al.  Variations in gene and protein expression in human nucleus pulposus in comparison with annulus fibrosus and cartilage cells: potential associations with aging and degeneration. , 2010, Osteoarthritis and cartilage.

[83]  John F. Bolton,et al.  Chondrocyte deformation within compressed agarose constructs at the cellular and sub-cellular levels. , 2000, Journal of biomechanics.

[84]  M. Revel,et al.  Modulation of proteoglycan production by cyclic tensile stretch in intervertebral disc cells through a post-translational mechanism. , 2006, Biorheology.

[85]  K. Ito,et al.  Directing bone marrow-derived stromal cell function with mechanics. , 2010, Journal of biomechanics.

[86]  B B Seedhom,et al.  Light and low-frequency pulsatile hydrostatic pressure enhances extracellular matrix formation by bone marrow mesenchymal cells: an in-vitro study with special reference to cartilage repair. , 2007, Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine.

[87]  George M. Wahba,et al.  Intervertebral Disc Cell Therapy for Regeneration: Mesenchymal Stem Cell Implantation in Rat Intervertebral Discs , 2004, Annals of Biomedical Engineering.

[88]  D. Bergel,et al.  Effect of static load on matrix synthesis rates in the intervertebral disc measured in vitro by a new perfusion technique , 1995, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[89]  J. Iatridis,et al.  Behavior of Mesenchymal Stem Cells in the Chemical Microenvironment of the Intervertebral Disc , 2008, Spine.

[90]  K. Yonenobu,et al.  Mechanical Stress-Induced Apoptosis of Endplate Chondrocytes in Organ-Cultured Mouse Intervertebral Discs: An Ex Vivo Study , 2003, Spine.

[91]  J. Lotz,et al.  Compression-induced degeneration of the intervertebral disc: an in vivo mouse model and finite-element study. , 1998, Spine.

[92]  H. Tsuji,et al.  Nitric Oxide Mediates the Change of Proteoglycan Synthesis in the Human Lumbar Intervertebral Disc in Response to Hydrostatic Pressure , 2001, Spine.

[93]  J. Buckwalter,et al.  Musculoskeletal soft-tissue aging : impact on mobility , 1993 .

[94]  Mehran Kasra,et al.  Frequency response of pig intervertebral disc cells subjected to dynamic hydrostatic pressure , 2006, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[95]  J Kraemer,et al.  Dynamic characteristics of the vertebral column, effects of prolonged loading. , 1985, Ergonomics.

[96]  A Shirazi-Adl,et al.  Computation of coupled diffusion of oxygen, glucose and lactic acid in an intervertebral disc. , 2007, Journal of biomechanics.

[97]  L. Claes,et al.  Mechanical Stimulation Alters Pleiotrophin and Aggrecan Expression by Human Intervertebral Disc Cells and Influences Their Capacity to Stimulate Endothelial Cell Migration , 2009, Spine.

[98]  Ingo Müller,et al.  Low physiologic oxygen tensions reduce proliferation and differentiation of human multipotent mesenchymal stromal cells , 2010, BMC Cell Biology.

[99]  W. Hutton,et al.  An In Vivo MRI Study of the Changes in Volume (and Fluid Content) of the Lumbar Intervertebral Disc After Overnight Bed Rest and During an 8-Hour Walking Protocol , 2002, Journal of spinal disorders & techniques.

[100]  I. Kiviranta,et al.  Effect of Running Exercise on Proteoglycans and Collagen Content in the Intervertebral Disc of Young Dogs , 1993, International Journal of Sports Medicine.

[101]  R. Huiskes,et al.  Biophysical stimuli on cells during tissue differentiation at implant interfaces , 1997 .

[102]  A. Grodzinsky,et al.  Dynamic compression stimulates proteoglycan synthesis by mesenchymal stem cells in the absence of chondrogenic cytokines. , 2009, Tissue engineering. Part A.

[103]  T. Malone,et al.  Effects of running on intervertebral disc height. , 1990, The Journal of orthopaedic and sports physical therapy.

[104]  H. Cheung,et al.  Cyclic compression maintains viability and induces chondrogenesis of human mesenchymal stem cells in fibrin gel scaffolds. , 2009, Stem cells and development.

[105]  Rocky S Tuan,et al.  Intervertebral disc cell response to dynamic compression is age and frequency dependent , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[106]  M. Adams,et al.  What is Intervertebral Disc Degeneration, and What Causes It? , 2006, Spine.

[107]  J. Urban,et al.  Differential expression level of cytokeratin 8 in cells of the bovine nucleus pulposus complicates the search for specific intervertebral disc cell markers , 2010, Arthritis research & therapy.

[108]  M. Alini,et al.  Differential response of human bone marrow stromal cells to either TGF-β1 or rhGDF-5 , 2011, European Spine Journal.

[109]  Stuart B Goodman,et al.  Dose- and time-dependent effects of cyclic hydrostatic pressure on transforming growth factor-beta3-induced chondrogenesis by adult human mesenchymal stem cells in vitro. , 2006, Tissue engineering.

[110]  J. Hassenpflug,et al.  The influence of oxygen and hydrostatic pressure on articular chondrocytes and adherent bone marrow cells in vitro. , 2004, Biorheology.

[111]  A. Mobasheri,et al.  INTEGRINS AND STRETCH ACTIVATED ION CHANNELS; PUTATIVE COMPONENTS OF FUNCTIONAL CELL SURFACE MECHANORECEPTORS IN ARTICULAR CHONDROCYTES , 2002, Cell biology international.

[112]  S. Thorpe,et al.  Low oxygen tension is a more potent promoter of chondrogenic differentiation than dynamic compression. , 2010, Journal of biomechanics.

[113]  Christopher R Jacobs,et al.  The role of mechanical signals in regulating chondrogenesis and osteogenesis of mesenchymal stem cells. , 2010, Birth defects research. Part C, Embryo today : reviews.

[114]  A. Maroudas,et al.  Swelling of the intervertebral disc in vitro. , 1981, Connective tissue research.

[115]  Lutz Claes,et al.  A three-dimensional collagen matrix as a suitable culture system for the comparison of cyclic strain and hydrostatic pressure effects on intervertebral disc cells. , 2005, Journal of neurosurgery. Spine.

[116]  J. Lotz,et al.  Intervertebral Disc Cell Death Is Dependent on the Magnitude and Duration of Spinal Loading , 2000, Spine.

[117]  P. Janmey The cytoskeleton and cell signaling: component localization and mechanical coupling. , 1998, Physiological reviews.

[118]  Jon D. Szafranski,et al.  Chondrocyte mechanotransduction: effects of compression on deformation of intracellular organelles and relevance to cellular biosynthesis. , 2004, Osteoarthritis and cartilage.

[119]  John Y-J Shyy,et al.  Intervertebral disc degeneration: the role of the mitochondrial pathway in annulus fibrosus cell apoptosis induced by overload. , 2004, The American journal of pathology.

[120]  G S Beaupré,et al.  Correlations between mechanical stress history and tissue differentiation in initial fracture healing , 1988, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[121]  Mauro Alini,et al.  In vivo remodeling of intervertebral discs in response to short‐ and long‐term dynamic compression , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[122]  T. Albert,et al.  Fibroblast Growth Factor-2 Maintains the Differentiation Potential of Nucleus Pulposus Cells In Vitro: Implications for Cell-Based Transplantation Therapy , 2007, Spine.

[123]  Keita Ito,et al.  The Combined Effects of Limited Nutrition and High-Frequency Loading on Intervertebral Discs With Endplates , 2010, Spine.

[124]  P. Czermak,et al.  Hydrodynamic Stimulation and Long Term Cultivation of Nucleus Pulposus Cells: A New Bioreactor System to Induce Extracellular Matrix Synthesis by Nucleus Pulposus Cells Dependent on Intermittent Hydrostatic Pressure , 2004, The International journal of artificial organs.

[125]  J. Iatridis,et al.  In Vitro Organ Culture of the Bovine Intervertebral Disc: Effects of Vertebral Endplate and Potential for Mechanobiology Studies , 2006, Spine.

[126]  J. Iatridis,et al.  Characterization of an in vitro intervertebral disc organ culture system , 2007, European Spine Journal.

[127]  I. Stokes,et al.  Different Effects of Static Versus Cyclic Compressive Loading on Rat Intervertebral Disc Height and Water Loss In Vitro , 2007, Spine.

[128]  J. Urban,et al.  Regulation of matrix synthesis rates by the ionic and osmotic environment of articular chondrocytes , 1993, Journal of cellular physiology.

[129]  Karin Wuertz,et al.  MSC response to pH levels found in degenerating intervertebral discs. , 2009, Biochemical and biophysical research communications.

[130]  Yubo Sun,et al.  Effects of Cyclic Compressive Loading on Chondrogenesis of Rabbit Bone‐Marrow Derived Mesenchymal Stem Cells , 2004, Stem cells.

[131]  Mauro Alini,et al.  The effects of short‐term load duration on anabolic and catabolic gene expression in the rat tail intervertebral disc , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.