Bioactive Stratified Polymer Ceramic-Hydrogel Scaffold for Integrative Osteochondral Repair

Due to the intrinsically poor repair potential of articular cartilage, injuries to this soft tissue do not heal and require clinical intervention. Tissue engineered osteochondral grafts offer a promising alternative for cartilage repair. The functionality and integration potential of these grafts can be further improved by the regeneration of a stable calcified cartilage interface. This study focuses on the design and optimization of a stratified osteochondral graft with biomimetic multi-tissue regions, including a pre-designed and pre-integrated interface region. Specifically, the scaffold based on agarose hydrogel and composite microspheres of polylactide-co-glycolide (PLGA) and 45S5 bioactive glass (BG) was fabricated and optimized for chondrocyte density and microsphere composition. It was observed that the stratified scaffold supported the region-specific co-culture of chondrocytes and osteoblasts which can lead to the production of three distinct yet continuous regions of cartilage, calcified cartilage and bone-like matrices. Moreover, higher cell density enhanced chondrogenesis and improved graft mechanical property over time. The PLGA-BG phase promoted chondrocyte mineralization potential and is required for the formation of a calcified interface and bone regions on the osteochondral graft. These results demonstrate the potential of the stratified scaffold for integrative cartilage repair and future studies will focus on scaffold optimization and in vivo evaluations.

[1]  D. Burr,et al.  Anatomy and physiology of the mineralized tissues: role in the pathogenesis of osteoarthrosis. , 2004, Osteoarthritis and cartilage.

[2]  J. Buckwalter,et al.  Articular Cartilage and Osteoarthritis , 1954 .

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

[4]  M. Harper.  Viscous isoamyl 2‐cyanoacrylate as an osseous adhesive in the repair of osteochondral osteotomies in rabbits , 1988, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[5]  P. Bullough.  The role of joint architecture in the etiology of arthritis. , 2004, Osteoarthritis and cartilage.

[6]  J Wang,et al.  Formation of biphasic constructs containing cartilage with a calcified zone interface. , 2007, Tissue engineering.

[7]  G. Ateshian,et al.  The role of cell seeding density and nutrient supply for articular cartilage tissue engineering with deformational loading. , 2003, Osteoarthritis and cartilage.

[8]  J. Mao,et al.  Tissue-engineered Neogenesis of Human-shaped Mandibular Condyle from Rat Mesenchymal Stem Cells , 2003, Journal of dental research.

[9]  K. Abendroth,et al.  Analyses of the tidemark on human femoral heads. I. Histochemical, ultrastructural and microanalytic characterization of the normal structure of the intercartilaginous junction. , 1989, Acta morphologica Hungarica.

[10]  Cato T Laurencin,et al.  Three-dimensional, bioactive, biodegradable, polymer-bioactive glass composite scaffolds with improved mechanical properties support collagen synthesis and mineralization of human osteoblast-like cells in vitro. , 2003, Journal of biomedical materials research. Part A.

[11]  J. Buckwalter,et al.  Interspecies comparisons of in situ intrinsic mechanical properties of distal femoral cartilage , 1991, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[12]  D. D’Lima,et al.  Impact of Mechanical Trauma on Matrix and Cells , 2001, Clinical orthopaedics and related research.

[13]  Magnus Lundberg,et al.  Treatment of osteochondral defects with autologous bone marrow in a hyaluronan-based delivery vehicle. , 2002, Tissue engineering.

[14]  S. Waldman,et al.  Repair of osteochondral defects with biphasic cartilage-calcium polyphosphate constructs in a sheep model. , 2006, Biomaterials.

[15]  Scott C. Brown,et al.  A three-dimensional osteochondral composite scaffold for articular cartilage repair. , 2002, Biomaterials.

[16]  Jeannine S. Schiller,et al.  Summary health statistics for U.S. adults: National Health Interview Survey, 2002. , 2004, Vital and health statistics. Series 10, Data from the National Health Survey.

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

[18]  I. Kanat,et al.  Subchondral bone drilling: a treatment for cartilage defects. , 1990, The Journal of foot surgery.

[19]  R Langer,et al.  In vitro generation of osteochondral composites. , 2000, Biomaterials.

[20]  P. Bullough,et al.  The morphology of the calcification front in articular cartilage. Its significance in joint function. , 1983, The Journal of bone and joint surgery. British volume.

[21]  Scott J Hollister,et al.  Engineered osteochondral grafts using biphasic composite solid free-form fabricated scaffolds. , 2004, Tissue engineering.

[22]  Helen H. Lu,et al.  Compositional effects on the formation of a calcium phosphate layer and the response of osteoblast-like cells on polymer-bioactive glass composites. , 2005, Biomaterials.

[23]  The pathology of articular and spinal diseases , 1949 .

[24]  R. Salter,et al.  The chondrogenic potential of free autogenous periosteal grafts for biological resurfacing of major full-thickness defects in joint surfaces under the influence of continuous passive motion. An experimental investigation in the rabbit. , 1986, The Journal of bone and joint surgery. American volume.

[25]  I. Shapiro,et al.  A rapid and ultrasensitive method for measurement of DNA, calcium and protein content, and alkaline phosphatase activity of chondrocyte cultures , 1995, Calcified Tissue International.

[26]  C. Enwemeka,et al.  A simplified method for the analysis of hydroxyproline in biological tissues. , 1996, Clinical biochemistry.

[27]  S. L. Sledge,et al.  Microfracture techniques in the treatment of osteochondral injuries. , 2001, Clinics in sports medicine.

[28]  Jie Jiang,et al.  Co-culture of osteoblasts and chondrocytes modulates cellular differentiation in vitro. , 2005, Biochemical and biophysical research communications.

[29]  A. Thambyah.  A hypothesis matrix for studying biomechanical factors associated with the initiation and progression of posttraumatic osteoarthritis. , 2005, Medical hypotheses.

[30]  R. Lemperg.  The subchondral bone plate of the femoral head in adult rabbits , 2004, Virchows Archiv A.

[31]  S. Akizuki,et al.  A new method of hemostasis for cementless total knee arthroplasty. , 1997, Bulletin (Hospital for Joint Diseases (New York, N.Y.)).

[32]  J. Mcclure,et al.  The tidemark of the chondro-osseous junction of the normal human knee joint , 2005, Journal of Molecular Histology.

[33]  Arnold I Caplan,et al.  Repair of osteochondral defect with tissue-engineered two-phase composite material of injectable calcium phosphate and hyaluronan sponge. , 2002, Tissue engineering.

[34]  Dietmar W Hutmacher,et al.  Repair of large articular osteochondral defects using hybrid scaffolds and bone marrow-derived mesenchymal stem cells in a rabbit model. , 2006, Tissue engineering.

[35]  L. Gibson,et al.  Design of a multiphase osteochondral scaffold III: Fabrication of layered scaffolds with continuous interfaces. , 2009, Journal of biomedical materials research. Part A.

[36]  R Kujat,et al.  Engineering of osteochondral tissue with bone marrow mesenchymal progenitor cells in a derivatized hyaluronan-gelatin composite sponge. , 1999, Tissue engineering.

[37]  T. Brown,et al.  Mechanical determinants of osteoarthrosis. , 1991, Seminars in arthritis and rheumatism.

[38]  Kanat Io,et al.  Subchondral bone drilling: a treatment for cartilage defects. , 1990 .

[39]  J H Brekke,et al.  Regeneration of articular cartilage--evaluation of osteochondral defect repair in the rabbit using multiphasic implants. , 2005, Osteoarthritis and cartilage.

[40]  P H Krebsbach,et al.  Tissue engineering osteochondral implants for temporomandibular joint repair. , 2005, Orthodontics & craniofacial research.

[41]  Gerard A. Ateshian,et al.  Influence of Seeding Density and Dynamic Deformational Loading on the Developing Structure/Function Relationships of Chondrocyte-Seeded Agarose Hydrogels , 2002, Annals of Biomedical Engineering.

[42]  Larry L. Hench,et al.  Bioceramics: From Concept to Clinic , 1991 .

[43]  V. Mow.  Biomechanics of articular cartilage , 1989 .

[44]  J. Landells,et al.  Histochemical Studies of Rheumatic Conditions , 1953, Annals of the rheumatic diseases.

[45]  J. Elisseeff,et al.  Advances in skeletal tissue engineering with hydrogels. , 2005, Orthodontics & craniofacial research.

[46]  Alan Grodzinsky,et al.  Tissue-engineered composites for the repair of large osteochondral defects. , 2002, Arthritis and rheumatism.

[47]  L. Hangody,et al.  Arthroscopic autogenous osteochondral mosaicplasty for the treatment of femoral condylar articular defects A preliminary report , 1997, Knee Surgery, Sports Traumatology, Arthroscopy.

[48]  D. Amiel,et al.  Integrative cartilage repair: Inhibition by β‐aminopropionitrile , 1999 .

[49]  Michael A. Slivka,et al.  Evaluation of multiphase implants for repair of focal osteochondral defects in goats. , 2000, Biomaterials.

[50]  H.W.J. Huiskes,et al.  Basic orthopaedic biomechanics and mechano-biology , 2005 .

[51]  E. Hunziker,et al.  Structural Barrier Principle for Growth Factor-Based Articular Cartilage Repair , 2001, Clinical orthopaedics and related research.

[52]  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.

[53]  Jie Jiang,et al.  Osteoblast and chondrocyte interactions during coculture on scaffolds. , 2003, IEEE engineering in medicine and biology magazine : the quarterly magazine of the Engineering in Medicine & Biology Society.

[54]  S. Havelka,et al.  The calcified-noncalcified cartilage interface: the tidemark. , 1984, Acta biologica Hungarica.

[55]  V. Mow,et al.  The ultrastructure and biomechanical significance of the tidemark of articular cartilage. , 1975, Clinical orthopaedics and related research.

[56]  Aldo R Boccaccini,et al.  Bioactive composite materials for tissue engineering scaffolds , 2005, Expert review of medical devices.

[57]  M. Knight,et al.  Mechanical compression influences intracellular Ca2+ signaling in chondrocytes seeded in agarose constructs. , 2001, Journal of applied physiology.

[58]  A I Caplan,et al.  Tissue-engineered fabrication of an osteochondral composite graft using rat bone marrow-derived mesenchymal stem cells. , 2001, Tissue engineering.

[59]  P. Benya,et al.  Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels , 1982, Cell.

[60]  C. Archer,et al.  Current strategies for articular cartilage repair. , 2005, European cells & materials.

[61]  Richard Tuli,et al.  Human mesenchymal progenitor cell-based tissue engineering of a single-unit osteochondral construct. , 2004, Tissue engineering.

[62]  D. Bader,et al.  Dynamic compression counteracts IL-1β-induced release of nitric oxide and PGE2by superficial zone chondrocytes cultured in agarose constructs , 2003 .

[63]  B. A. Byers,et al.  The beneficial effect of delayed compressive loading on tissue-engineered cartilage constructs cultured with TGF-beta3. , 2007, Osteoarthritis and cartilage.

[64]  Sandra L. Johnson,et al.  PROSTAGLANDINS AND THE ZONE OF CALCIFIED CARTILAGE IN OSTEOARTHRITIS , 1996, American Journal of Therapeutics.

[65]  T. Oegema,et al.  The interaction of the zone of calcified cartilage and subchondral bone in osteoarthritis , 1997, Microscopy research and technique.

[66]  S. Teoh,et al.  Scaffold design and in vitro study of osteochondral coculture in a three-dimensional porous polycaprolactone scaffold fabricated by fused deposition modeling. , 2003, Tissue engineering.

[67]  D. W. Jackson,et al.  Cartilage Substitutes: Overview of Basic Science and Treatment Options , 2001, The Journal of the American Academy of Orthopaedic Surgeons.

[68]  Van C. Mow,et al.  Structure and function of articular cartilage and meniscus , 2005 .

[69]  G. Ateshian,et al.  Scaffold degradation elevates the collagen content and dynamic compressive modulus in engineered articular cartilage. , 2009, Osteoarthritis and cartilage.

[70]  P. Bullough,et al.  The geometry of diarthrodial joints, its physiologic maintenance, and the possible significance of age-related changes in geometry-to-load distribution and the development of osteoarthritis. , 1981, Clinical orthopaedics and related research.

[71]  Antonios G Mikos,et al.  Osteochondral repair in the rabbit model utilizing bilayered, degradable oligo(poly(ethylene glycol) fumarate) hydrogel scaffolds. , 2005, Journal of biomedical materials research. Part A.

[72]  D. Amiel,et al.  Integrative cartilage repair: inhibition by beta-aminopropionitrile. , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[73]  G. Ateshian,et al.  Anatomically shaped osteochondral constructs for articular cartilage repair. , 2003, Journal of biomechanics.

[74]  Jeremy J Mao,et al.  Tissue-engineered osteochondral constructs in the shape of an articular condyle. , 2005, The Journal of bone and joint surgery. American volume.

[75]  Liming Bian,et al.  Passaged adult chondrocytes can form engineered cartilage with functional mechanical properties: a canine model. , 2010, Tissue engineering. Part A.

[76]  Helen H. Lu,et al.  Controlled delivery of platelet-rich plasma-derived growth factors for bone formation. , 2008, Journal of biomedical materials research. Part A.

[77]  E B Hunziker,et al.  Functional barrier principle for growth-factor-based articular cartilage repair. , 2003, Osteoarthritis and cartilage.

[78]  P. Bullough,et al.  Age-related changes in the thickness of the calcified zone and the number of tidemarks in adult human articular cartilage. , 1980, The Journal of bone and joint surgery. British volume.

[79]  Margareta Nordin,et al.  Basic Biomechanics of the Musculoskeletal Systm , 1989 .

[80]  M. Ochi,et al.  Current concepts in tissue engineering technique for repair of cartilage defect. , 2001, Artificial organs.

[81]  V. Goldberg,et al.  Repair of osteochondral defects with hyaluronan- and polyester-based scaffolds. , 2005, Osteoarthritis and cartilage.