Mechanics of oriented electrospun nanofibrous scaffolds for annulus fibrosus tissue engineering

Engineering a functional replacement for the annulus fibrosus (AF) of the intervertebral disc is contingent upon recapitulation of AF structure, composition, and mechanical properties. In this study, we propose a new paradigm for AF tissue engineering that focuses on the reconstitution of anatomic fiber architecture and uses constitutive modeling to evaluate construct function. A modified electrospinning technique was utilized to generate aligned nanofibrous polymer scaffolds for engineering the basic functional unit of the AF, a single lamella. Scaffolds were tested in uniaxial tension at multiple fiber orientations, demonstrating a nonlinear dependence of modulus on fiber angle that mimicked the nonlinearity and anisotropy of native AF. A homogenization model previously applied to native AF successfully described scaffold mechanical response, and parametric studies demonstrated that nonfibrillar matrix, along with fiber connectivity, are key contributors to tensile mechanics for engineered AF. We demonstrated that AF cells orient themselves along the aligned scaffolds and deposit matrix that contributes to construct mechanics under loading conditions relevant to the in vivo environment. The homogenization model was applied to cell‐seeded constructs and provided quantitative measures for the evolution of matrix and interfibrillar interactions. Finally, the model demonstrated that at fiber angles of the AF (28°–44°), engineered material behaved much like native tissue, suggesting that engineered constructs replicate the physiologic behavior of the single AF lamella. Constitutive modeling provides a powerful tool for analysis of engineered AF neo‐tissue and native AF tissue alike, highlighting key mechanical design criteria for functional AF tissue engineering. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1018–1028, 2007

[1]  Jun Hu,et al.  Alignment of osteoblast-like cells and cell-produced collagen matrix induced by nanogrooves. , 2005, Tissue engineering.

[2]  Gerhard A. Holzapfel,et al.  An Anisotropic Model for Annulus Tissue and Enhanced Finite Element Analyses of Intact Lumbar Disc Bodies , 2001 .

[3]  L. Setton,et al.  A linear material model for fiber-induced anisotropy of the anulus fibrosus. , 2000, Journal of biomechanical engineering.

[4]  Delphine Périé,et al.  Confined compression experiments on bovine nucleus pulposus and annulus fibrosus: sensitivity of the experiment in the determination of compressive modulus and hydraulic permeability. , 2005, Journal of biomechanics.

[5]  V. C. Mow,et al.  Regional Variation in Tensile Properties and Biochemical Composition of the Human Lumbar Anulus Fibrosus , 1994, Spine.

[6]  D. S. Hickey,et al.  X-ray diffraction studies of the arrangement of collagenous fibres in human fetal intervertebral disc. , 1980, Journal of anatomy.

[7]  A. Hiltner,et al.  Hierarchical structure of the intervertebral disc. , 1989, Connective tissue research.

[8]  V C Mow,et al.  Tensile Properties of Nondegenerate Human Lumbar Anulus Fibrosus , 1996, Spine.

[9]  Wan-Ju Li,et al.  Electrospun Nanofibrous Scaffolds: Production, Characterization, and Applications for Tissue Engineering and Drug Delivery , 2005 .

[10]  H. Wu,et al.  Mechanical behavior of the human annulus fibrosus. , 1976, Journal of biomechanics.

[11]  Van C. Mow,et al.  Degeneration and Aging Affect the Tensile Behavior of Human Lumbar Anulus Fibrosus , 1995, Spine.

[12]  V C Mow,et al.  Shear mechanical properties of human lumbar annulus fibrosus , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[13]  Jeffrey C Lotz,et al.  Theoretical model and experimental results for the nonlinear elastic behavior of human annulus fibrosus , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[14]  L. Setton,et al.  Collagen gene expression and mechanical properties of intervertebral disc cell–alginate cultures , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[15]  D. Eyre,et al.  Biochemistry of the intervertebral disc. , 1979, International review of connective tissue research.

[16]  L. Setton,et al.  Anisotropic and inhomogeneous tensile behavior of the human anulus fibrosus: experimental measurement and material model predictions. , 2001, Journal of biomechanical engineering.

[17]  J. A. Cooper,et al.  Fabrication and characterization of six electrospun poly(alpha-hydroxy ester)-based fibrous scaffolds for tissue engineering applications. , 2006, Acta biomaterialia.

[18]  H. Gruber,et al.  Three-dimensional culture of human disc cells within agarose or a collagen sponge: assessment of proteoglycan production. , 2006, Biomaterials.

[19]  J. Urban,et al.  Cells From Different Regions of the Intervertebral Disc: Effect of Culture System on Matrix Expression and Cell Phenotype , 2002, Spine.

[20]  L. Dahners,et al.  Collagen fiber sliding during ligament growth and contracture , 1998, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  T. Oegema,et al.  Biochemistry of the intervertebral disc. , 1993, Clinics in sports medicine.

[22]  M. Aebi,et al.  The Potential and Limitations of a Cell-Seeded Collagen/Hyaluronan Scaffold to Engineer an Intervertebral Disc-Like Matrix , 2003, Spine.

[23]  S. Roberts,et al.  Human intervertebral disc cell morphology and cytoskeletal composition: a preliminary study of regional variations in health and disease , 2003, Journal of anatomy.

[24]  S. Bruehlmann,et al.  Regional variations in the cellular matrix of the annulus fibrosus of the intervertebral disc , 2002, Journal of anatomy.

[25]  J. Ralphs,et al.  The development of fibrocartilage in the rat intervertebral disc , 1995, Anatomy and Embryology.

[26]  P. Hulme,et al.  In situ intercellular mechanics of the bovine outer annulus fibrosus subjected to biaxial strains. , 2004, Journal of biomechanics.

[27]  M. Segal,et al.  Biaxial Testing of Human Annulus Fibrosus and Its Implications for a Constitutive Formulation , 2004, Annals of Biomedical Engineering.

[28]  P. Czermak,et al.  A Stimulation Unit for the Application of Mechanical Strain on Tissue Engineered Anulus Fibrosus Cells: A New System to Induce Extracellular Matrix Synthesis by Anulus Fibrosus Cells Dependent on Cyclic Mechanical Strain , 2005, The International journal of artificial organs.

[29]  Luzhong Yin,et al.  A homogenization model of the annulus fibrosus. , 2005, Journal of biomechanics.

[30]  J. Ralphs,et al.  Role of actin stress fibres in the development of the intervertebral disc: Cytoskeletal control of extracellular matrix assembly , 1999, Developmental dynamics : an official publication of the American Association of Anatomists.

[31]  Heather L Guerin,et al.  Quantifying the contributions of structure to annulus fibrosus mechanical function using a nonlinear, anisotropic, hyperelastic model , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[32]  M. Spector,et al.  Effects of collagen type on the behavior of adult canine annulus fibrosus cells in collagen-glycosaminoglycan scaffolds. , 2004, Journal of biomedical materials research. Part A.

[33]  M. Ishihara,et al.  Tissue engineering of the intervertebral disc with cultured annulus fibrosus cells using atelocollagen honeycombshaped scaffold with a membrane seal (ACHMS scaffold) , 2003, Medical and Biological Engineering and Computing.

[34]  L. Bonassar,et al.  Biomechanical and biochemical characterization of composite tissue-engineered intervertebral discs. , 2006, Biomaterials.

[35]  P. Regitnig,et al.  Single lamellar mechanics of the human lumbar anulus fibrosus , 2005, Biomechanics and modeling in mechanobiology.

[36]  E. Thonar,et al.  Metabolism of the Extracellular Matrix Formed by Intervertebral Disc Cells Cultured in Alginate , 1997, Spine.

[37]  J. A. Cooper,et al.  Engineering controllable anisotropy in electrospun biodegradable nanofibrous scaffolds for musculoskeletal tissue engineering. , 2007, Journal of biomechanics.

[38]  T. Oegema,et al.  Identification of heterogeneous cell populations in normal human intervertebral disc. , 1995, Journal of anatomy.

[39]  Savio L-Y Woo,et al.  Cell orientation determines the alignment of cell-produced collagenous matrix. , 2003, Journal of biomechanics.

[40]  L. Setton,et al.  Intervertebral Disc Cells Exhibit Differences in Gene Expression in Alginate and Monolayer Culture , 2001, Spine.

[41]  H. Gruber,et al.  Cell-based tissue engineering for the intervertebral disc: in vitro studies of human disc cell gene expression and matrix production within selected cell carriers. , 2004, The spine journal : official journal of the North American Spine Society.

[42]  R. Tuan,et al.  Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in long-term agarose culture. , 2006, Osteoarthritis and cartilage.

[43]  F. Marchand,et al.  Investigation of the Laminate Structure of Lumbar Disc Anulus Fibrosus , 1990, Spine.

[44]  Michael S Sacks,et al.  Design and analysis of tissue engineering scaffolds that mimic soft tissue mechanical anisotropy. , 2006, Biomaterials.

[45]  S. Bruehlmann,et al.  ISSLS Prize Winner: Collagen Fibril Sliding Governs Cell Mechanics in the Anulus Fibrosus: An In Situ Confocal Microscopy Study of Bovine Discs , 2004, Spine.

[46]  C. Lim,et al.  Tensile testing of a single ultrafine polymeric fiber. , 2005, Biomaterials.