Characterization of a Novel Fiber Composite Material for Mechanotransduction Research of Fibrous Connective Tissues

Mechanotransduction is the fundamental process by which cells detect and respond to their mechanical environment, and is critical for tissue homeostasis. Understanding mechanotransduction mechanisms will provide insights into disease processes and injuries, and may support novel tissue engineering research. Although there has been extensive research in mechanotransduction, many pathways remain unclear, due to the complexity of the signaling mechanisms and loading environments involved. This study describes the development of a novel hydrogel-based fiber composite material for investigating mechanotransduction in fibrous tissues. By encapsulating poly(2-hydroxyethyl methacrylate) rods in a bulk poly(ethylene glycol) matrix, it aims to create a micromechanical environment more representative of that seen in vivo. Results demonstrated that collagen-coated rods enable localized cell attachment, and cells are successfully cultured for one week within the composite. Mechanical analysis of the composite indicates that gross mechanical properties and local strain environments could be manipulated by altering the fabrication process. Allowing diffusion between the rods and surrounding matrix creates an interpenetrating network whereby the relationships between shear and tension are altered. Increasing diffusion enhances the shear bond strength between rods and matrix and the levels of local tension along the rods. Preliminary investigation into fibroblast mechanotransduction illustrates that the fiber composite upregulates collagen I expression, the main protein in fibrous tissues, in response to cyclic tensile strains when compared to less complex 2D and 3D environments. In summary, the ability to create and manipulate a strain environment surrounding the fibers, where combined tensile and shear forces uniquely impact cell functions, is demonstrated.

[1]  D. A. Lee,et al.  Cyclic tensile strain upregulates collagen synthesis in isolated tendon fascicles. , 2005, Biochemical and biophysical research communications.

[2]  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.

[3]  S. Arnoczky,et al.  In situ cell nucleus deformation in tendons under tensile load; a morphological analysis using confocal laser microscopy , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[4]  S. Arnoczky,et al.  Effect of Amplitude and Frequency of Cyclic Tensile Strain on the Inhibition of MMP-1 mRNA Expression in Tendon Cells: An In Vitro Study , 2003, Connective tissue research.

[5]  Michael Lavagnino,et al.  Ex vivo static tensile loading inhibits MMP‐1 expression in rat tail tendon cells through a cytoskeletally based mechanotransduction mechanism , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[6]  L. Griffith,et al.  Capturing complex 3D tissue physiology in vitro , 2006, Nature Reviews Molecular Cell Biology.

[7]  M. Knight,et al.  Loading alters actin dynamics and up-regulates cofilin gene expression in chondrocytes. , 2007, Biochemical and biophysical research communications.

[8]  S. Bryant,et al.  The role of hydrogel structure and dynamic loading on chondrocyte gene expression and matrix formation. , 2008, Journal of biomechanics.

[9]  U Bosch,et al.  The Proliferative Response of Isolated Human Tendon Fibroblasts to Cyclic Biaxial Mechanical Strain * , 2000, The American journal of sports medicine.

[10]  P. Fratzl,et al.  Viscoelastic properties of collagen: synchrotron radiation investigations and structural model. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[11]  K. Pal,et al.  Polymeric Hydrogels: Characterization and Biomedical Applications , 2009 .

[12]  N. Sasaki,et al.  Elongation mechanism of collagen fibrils and force-strain relations of tendon at each level of structural hierarchy. , 1996, Journal of biomechanics.

[13]  D. Bader,et al.  An investigation into the effects of the hierarchical structure of tendon fascicles on micromechanical properties , 2004, Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine.

[14]  Aaditya C Devkota,et al.  Cyclic loading alters biomechanical properties and secretion of PGE2 and NO from tendon explants. , 2006, Clinical biomechanics.

[15]  Carol A. Otey,et al.  Mechanical forces and signaling in connective tissue cells: Cellular mechanisms of detection, transduction, and responses to mechanical deformation , 2001 .

[16]  D L Bader,et al.  In vitro fatigue of human tendons. , 1997, Journal of biomechanics.

[17]  D L Bader,et al.  Development of a technique to determine strains in tendons using the cell nuclei. , 2003, Biorheology.

[18]  Shu Chien,et al.  Mechanotransduction in Response to Shear Stress , 1999, The Journal of Biological Chemistry.

[19]  Joseph W Freeman,et al.  Collagen self-assembly and the development of tendon mechanical properties. , 2003, Journal of biomechanics.

[20]  D. A. Lee,et al.  Differential regulation of gene expression in isolated tendon fascicles exposed to cyclic tensile strain in vitro. , 2009, Journal of applied physiology.

[21]  H. Screen,et al.  The micro-structural strain response of tendon , 2007 .

[22]  M. Koch,et al.  Stress-induced molecular rearrangement in tendon collagen. , 1985, Journal of molecular biology.

[23]  A Hosseini,et al.  In-vivo time-dependent articular cartilage contact behavior of the tibiofemoral joint. , 2010, Osteoarthritis and cartilage.

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

[25]  S. Bryant,et al.  Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels. , 2002, Journal of biomedical materials research.

[26]  M. van Griensven,et al.  Modulation of cell functions of human tendon fibroblasts by different repetitive cyclic mechanical stress patterns. , 2003, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[27]  S J Bryant,et al.  Static and dynamic compressive strains influence nitric oxide production and chondrocyte bioactivity when encapsulated in PEG hydrogels of different crosslinking densities. , 2008, Osteoarthritis and cartilage.

[28]  A. Khademhosseini,et al.  Hydrogels in Regenerative Medicine , 2009, Advanced materials.

[29]  D. Butler,et al.  Tensile stimulation of murine stem cell-collagen sponge constructs increases collagen type I gene expression and linear stiffness. , 2009, Tissue engineering. Part A.

[30]  Kristi S. Anseth,et al.  PEG Hydrogels for the Controlled Release of Biomolecules in Regenerative Medicine , 2009, Pharmaceutical Research.

[31]  A. Banes,et al.  ATP modulates load‐inducible IL‐1β, COX 2, and MMP‐3 gene expression in human tendon cells , 2003, Journal of cellular biochemistry.

[32]  S J Bryant,et al.  Cytocompatibility of UV and visible light photoinitiating systems on cultured NIH/3T3 fibroblasts in vitro , 2000, Journal of biomaterials science. Polymer edition.

[33]  R. F. Ker Mechanics of tendon, from an engineering perspective , 2007 .

[34]  W. Herzog,et al.  Stretch and interleukin‐1β induce matrix metalloproteinases in rabbit tendon cells in vitro , 2002 .

[35]  Jiandong Ding,et al.  A Macroscopic Helix Formation Induced by the Shrinking of a Cylindrical Polymeric Hydrogel , 2001 .

[36]  J. P. Paul,et al.  In vivo human tendon mechanical properties , 1999, The Journal of physiology.

[37]  B. Baroli,et al.  Hydrogels for tissue engineering and delivery of tissue-inducing substances. , 2007, Journal of pharmaceutical sciences.

[38]  M. Hendzel,et al.  Mechanotransduction from the ECM to the genome: Are the pieces now in place? , 2008, Journal of cellular biochemistry.

[39]  S. Bryant,et al.  Cell encapsulation in biodegradable hydrogels for tissue engineering applications. , 2008, Tissue engineering. Part B, Reviews.

[40]  N. Sasaki,et al.  Stress-strain curve and Young's modulus of a collagen molecule as determined by the X-ray diffraction technique. , 1996, Journal of biomechanics.

[41]  J E Scott,et al.  Elasticity in extracellular matrix ‘shape modules’ of tendon, cartilage, etc. A sliding proteoglycan‐filament model , 2003, The Journal of physiology.

[42]  Natalia Juncosa-Melvin,et al.  Mechanical stimulation increases collagen type I and collagen type III gene expression of stem cell-collagen sponge constructs for patellar tendon repair. , 2007, Tissue engineering.

[43]  R. Zernicke,et al.  Mechanical load stimulates expression of novel genes in vivo and in vitro in avian flexor tendon cells. , 1999, Osteoarthritis and cartilage.

[44]  A. Khademhosseini,et al.  Hydrogels in Biology and Medicine: From Molecular Principles to Bionanotechnology , 2006 .

[45]  Lin Yu,et al.  Injectable hydrogels as unique biomedical materials. , 2008, Chemical Society reviews.