Type II collagen-hyaluronan hydrogel--a step towards a scaffold for intervertebral disc tissue engineering.

Intervertebral disc regeneration strategies based on stem cell differentiation in combination with the design of functional scaffolds is an attractive approach towards repairing/regenerating the nucleus pulposus. The specific aim of this study was to optimise a composite hydrogel composed of type II collagen and hyaluronic acid (HA) as a carrier for mesenchymal stem cells. Hydrogel stabilisation was achieved by means of 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) cross-linking. Optimal hydrogel properties were determined by investigating different concentrations of EDC (8 mM, 24 mM and 48 mM). Stable hydrogels were obtained independent of the concentration of carbodiimide used. The hydrogels cross-linked by the lowest concentration of EDC (8 mM) demonstrated high swelling properties. Additionally, improved proliferation of seeded rat mesenchymal stem cells (rMSCs) and hydrogel stability levels in culture were observed with this 8 mM cross-linked hydrogel. Results from this study indicate that EDC/NHS (8 mM) cross-linked type II collagen/HA hydrogel was capable of supporting viability of rMSCs, and furthermore their differentiation into a chondrogenic lineage. Further investigations should be conducted to determine its potential as scaffold for nucleus pulposus regeneration/repair.

[1]  R. Bank,et al.  Influence of collagen type II and nucleus pulposus cells on aggregation and differentiation of adipose tissue-derived stem cells , 2008, Journal of cellular and molecular medicine.

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

[3]  N. Takahashi,et al.  Hyaluronan promotes the chondrocyte response to BMP-7. , 2009, Osteoarthritis and cartilage.

[4]  A Ratcliffe,et al.  Determination of collagen-proteoglycan interactions in vitro. , 1996, Journal of biomechanics.

[5]  T. Hardingham The role of link-protein in the structure of cartilage proteoglycan aggregates. , 1979, The Biochemical journal.

[6]  浅沼 広子,et al.  High-Throughput な迅速凍結標本の作製 , 2002 .

[7]  J. Cleland,et al.  Physical Therapy for Acute Low Back Pain: Associations With Subsequent Healthcare Costs , 2008, Spine.

[8]  K. Ando,et al.  Atelocollagen for culture of human nucleus pulposus cells forming nucleus pulposus-like tissue in vitro: influence on the proliferation and proteoglycan production of HNPSV-1 cells. , 2006, Biomaterials.

[9]  B. Keil,et al.  Differences in the degradation of native collagen by two microbial collagenases. , 1979, The Biochemical journal.

[10]  Wan-Ju Li,et al.  Intervertebral disc tissue engineering using a novel hyaluronic acid-nanofibrous scaffold (HANFS) amalgam. , 2008, Tissue engineering. Part A.

[11]  Y. Ikada,et al.  Mechanism of amide formation by carbodiimide for bioconjugation in aqueous media. , 1995, Bioconjugate chemistry.

[12]  A. Pandit,et al.  Assessment of cell viability in a three-dimensional enzymatically cross-linked collagen scaffold , 2007, Journal of materials science. Materials in medicine.

[13]  D. Eyre,et al.  Collagen polymorphisms of the intervertebral disc. , 2002, Biochemical Society transactions.

[14]  M. Spector,et al.  Fabrication and characterization of porous hyaluronic acid-collagen composite scaffolds. , 2007, Journal of biomedical materials research. Part A.

[15]  W. Cats-Baril,et al.  An overview of the incidences and costs of low back pain. , 1991, The Orthopedic clinics of North America.

[16]  N. Ashammakhi,et al.  Effect of human platelet supernatant on proliferation and matrix synthesis of human articular chondrocytes in monolayer and three-dimensional alginate cultures. , 2005, Biomaterials.

[17]  A. Kjøniksen,et al.  Characterization of the chemical degradation of hyaluronic acid during chemical gelation in the presence of different cross-linker agents. , 2007, Carbohydrate research.

[18]  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.

[19]  Sally Roberts,et al.  Histology and pathology of the human intervertebral disc. , 2006, The Journal of bone and joint surgery. American volume.

[20]  T. Taguchi,et al.  An improved method to prepare hyaluronic acid and type II collagen composite matrices. , 2002, Journal of biomedical materials research.

[21]  S. Ayad,et al.  1991 Volvo Award in Basic Sciences: Collagen Types Around the Cells of the Intervertebral Disc and Cartilage End Plate: An Immunolocalization Study , 1991, Spine.

[22]  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.

[23]  J. M. Lee,et al.  Control of pH alters the type of cross-linking produced by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) treatment of acellular matrix vascular grafts. , 2001, Journal of biomedical materials research.

[24]  W. B. van den Berg,et al.  Crosslinked type II collagen matrices: preparation, characterization, and potential for cartilage engineering. , 2002, Biomaterials.

[25]  A. Pandit,et al.  Characterization of a microbial transglutaminase cross-linked type II collagen scaffold. , 2006, Tissue engineering.

[26]  C. Perka,et al.  Cultivation of porcine cells from the nucleus pulposus in a fibrin/hyaluronic acid matrix , 2000, Acta orthopaedica Scandinavica.

[27]  J. Matyas,et al.  The notochordal cell in the nucleus pulposus: a review in the context of tissue engineering. , 2003, Tissue engineering.

[28]  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.

[29]  A Rajaram,et al.  Influence of different crosslinking treatments on the physical properties of collagen membranes. , 2003, Biomaterials.

[30]  T. Strine,et al.  US national prevalence and correlates of low back and neck pain among adults. , 2007, Arthritis and rheumatism.

[31]  David L Kaplan,et al.  Stem cell- and scaffold-based tissue engineering approaches to osteochondral regenerative medicine. , 2009, Seminars in cell & developmental biology.

[32]  N. Adachi,et al.  Hyaluronic acid enhances proliferation and chondroitin sulfate synthesis in cultured chondrocytes embedded in collagen gels , 1999, Journal of cellular physiology.

[33]  G. Banfi,et al.  Pathophysiology of the human intervertebral disc. , 2008, The international journal of biochemistry & cell biology.

[34]  P. Roughley,et al.  The potential of chitosan-based gels containing intervertebral disc cells for nucleus pulposus supplementation. , 2006, Biomaterials.

[35]  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.

[36]  Frymoyer Jw,et al.  An overview of the incidences and costs of low back pain. , 1991 .

[37]  Mansoor M. Amiji,et al.  Preparation and Characterization of Freeze-dried Chitosan-Poly(Ethylene Oxide) Hydrogels for Site-Specific Antibiotic Delivery in the Stomach , 1996, Pharmaceutical Research.

[38]  E. Thonar,et al.  A New Culture System to Study the Metabolism of the Intervertebral Disc In Vitro , 1998, Spine.

[39]  M. Risbud,et al.  Properties of polyvinyl pyrrolidone/β-chitosan hydrogel membranes and their biocompatibility evaluation by haemorheological method , 2001, Journal of materials science. Materials in medicine.

[40]  Gianluca Gallo,et al.  Reevaluation of in vitro differentiation protocols for bone marrow stromal cells: Disruption of actin cytoskeleton induces rapid morphological changes and mimics neuronal phenotype , 2004, Journal of neuroscience research.

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

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

[43]  R. Soames,et al.  Human intervertebral disc: Structure and function , 1988, The Anatomical record.

[44]  Dawn M. Elliott,et al.  Material properties in unconfined compression of human nucleus pulposus, injectable hyaluronic acid-based hydrogels and tissue engineering scaffolds , 2007, European Spine Journal.

[45]  M. Alini,et al.  An injectable cross-linked scaffold for nucleus pulposus regeneration. , 2008, Biomaterials.

[46]  L G Griffith,et al.  Effect of pore size and void fraction on cellular adhesion, proliferation, and matrix deposition. , 2001, Tissue engineering.

[47]  A. Tengblad,et al.  Demonstration of link protein in proteoglycan aggregates from human intervertebral disc. , 1984, The Biochemical journal.

[48]  Sally Roberts,et al.  Degeneration of the intervertebral disc , 2003, Arthritis research & therapy.

[49]  K. Kelnar,et al.  High-throughput RNAi screening in vitro: from cell lines to primary cells. , 2005, RNA.

[50]  T. Hardingham,et al.  Structure and interactions of cartilage proteoglycan binding region and link protein. , 1985, The Biochemical journal.

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

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

[53]  James D. Kang,et al.  Feasibility of a stem cell therapy for intervertebral disc degeneration. , 2008, The spine journal : official journal of the North American Spine Society.

[54]  Shiying Xu,et al.  EDC/NHS-crosslinked type II collagen-chondroitin sulfate scaffold: characterization and in vitro evaluation , 2008, Journal of materials science. Materials in medicine.

[55]  Tatiana Segura,et al.  Crosslinked hyaluronic acid hydrogels: a strategy to functionalize and pattern. , 2005, Biomaterials.

[56]  C. V. van Blitterswijk,et al.  Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[57]  G. Bowlin,et al.  Cross-linking electrospun type II collagen tissue engineering scaffolds with carbodiimide in ethanol. , 2007, Tissue engineering.

[58]  James D. Kang,et al.  Biochemistry of intervertebral disc degeneration and the potential for gene therapy applications. , 2001, The spine journal : official journal of the North American Spine Society.

[59]  C. Laurencin,et al.  Biphasic scaffold for annulus fibrosus tissue regeneration. , 2008, Biomaterials.

[60]  M. Pajic,et al.  Nucleus Pulposus Cellular Longevity by Telomerase Gene Therapy , 2007, Spine.

[61]  T. Taguchi,et al.  Characterization of alkali-treated collagen gels prepared by different crosslinkers , 2008, Journal of materials science. Materials in medicine.

[62]  W. Richter,et al.  The use of mesenchymal stem cells for chondrogenesis. , 2008, Injury.

[63]  Benjamin M Wu,et al.  Effect of scaffold architecture and pore size on smooth muscle cell growth. , 2008, Journal of biomedical materials research. Part A.

[64]  Farshid Guilak,et al.  Isolation of adipose-derived stem cells and their induction to a chondrogenic phenotype , 2010, Nature Protocols.

[65]  A. Freemont,et al.  Human mesenchymal stem cell differentiation to NP-like cells in chitosan-glycerophosphate hydrogels. , 2008, Biomaterials.

[66]  Myron Spector,et al.  Incorporation of hyaluronic acid into collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis , 2007, Biomedical materials.