Developing an Articular Cartilage Decellularization Process Toward Facet Joint Cartilage Replacement

OBJECTIVEThe facet joint has been identified as a significant source of morbidity in lower back pain. In general, treatments have focused on reducing the pain associated with facet joint osteoarthritis, and no treatments have targeted the development of a replacement tissue for arthritic facet articular cartilage. Therefore, the objective of this study was to develop a nonimmunogenic decellularized articular cartilage replacement tissue while maintaining functional properties similar to native facet cartilage tissue. METHODSIn vitro testing was performed on bovine articular cartilage explants. The effects of 2% sodium dodecyl sulfate (SDS), a detergent used for cell and nuclear membrane solubilization, on cartilage cellularity, biochemical, and biomechanical properties, were examined. Compressive biomechanical properties were determined using creep indentation, and the tensile biomechanical properties were obtained with uniaxial tensile testing. Biochemical assessment involved determination of the DNA content, glycosaminoglycan (GAG) content, and collagen content. Histological examination included hematoxylin and eosin staining for tissue cellularity, as well as staining for collagen and GAG. RESULTSTreatment with 2% SDS for 2 hours maintained the compressive and tensile biomechanical properties, as well as the GAG and collagen content while resulting in a decrease in cell nuclei and a 4% decrease in DNA content. Additionally, treatment for 8 hours resulted in complete histological decellularization and a 40% decrease in DNA content while maintaining collagen content and tensile properties. However, a significant decrease in compressive properties and GAG content was observed. Similar results were observed with 4 hours of treatment, although the decrease in DNA content was not as great as with 8 hours of treatment. CONCLUSIONTreatment with 2% SDS for 8 hours resulted in complete histological decellularization with decreased mechanical properties, whereas treatment for 2 hours maintained mechanical properties, but had a minimal effect on DNA content. Therefore, future studies must be performed to optimize a treatment for decellularization while maintaining mechanical properties close to those of facet joint cartilage. This study served as a step in creating a decellularized articular cartilage replacement tissue that could be used as a treatment for facet cartilage osteoarthritis.

[1]  F. J. Dzida,et al.  Comparative study of the intrinsic mechanical properties of the human acetabular and femoral head cartilage , 1991, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[2]  G. Ayala,et al.  Porcine cartilage transplants in the cynomolgus monkey. III. Transplantation of alpha-galactosidase-treated porcine cartilage. , 1998, Transplantation.

[3]  W. Hayes,et al.  A mathematical analysis for indentation tests of articular cartilage. , 1972, Journal of biomechanics.

[4]  Daniel H. Kim,et al.  Biomechanical, biochemical, and histological characterization of canine lumbar facet joint cartilage. , 2009, Journal of neurosurgery. Spine.

[5]  Jeffrey C Lotz,et al.  Glycation increases human annulus fibrosus stiffness in both experimental measurements and theoretical predictions. , 2006, Journal of biomechanics.

[6]  B. Walker,et al.  The prevalence of low back pain: a systematic review of the literature from 1966 to 1998. , 2000, Journal of spinal disorders.

[7]  M. Dunn,et al.  Effect of chemical treatments on tendon cellularity and mechanical properties. , 2000, Journal of biomedical materials research.

[8]  Jerry C. Hu,et al.  A self-assembling process in articular cartilage tissue engineering. , 2006, Tissue engineering.

[9]  T. Wick,et al.  Effect of low oxygen tension on tissue-engineered cartilage construct development in the concentric cylinder bioreactor. , 2004, Tissue engineering.

[10]  T. Hedman,et al.  Effects of exogenous crosslinking on in vitro tensile and compressive moduli of lumbar intervertebral discs. , 2007, Clinical biomechanics.

[11]  King H. Yang,et al.  Mechanism of facet load transmission as a hypothesis for low-back pain. , 1984, Spine.

[12]  Kyriacos A. Athanasiou,et al.  Erratum: Biomechanical properties of hip cartilage in experimental animal models (Clinical Orthopaedics and Related Research (1995) 316 (254-266)) , 1995 .

[13]  J. Platt,et al.  Immunopathology of hyperacute xenograft rejection in a swine-to-primate model. , 1991, Transplantation.

[14]  L. Rosenberg Chemical basis for the histological use of safranin O in the study of articular cartilage. , 1971, The Journal of bone and joint surgery. American volume.

[15]  Leonid Kalichman,et al.  Lumbar facet joint osteoarthritis: a review. , 2007, Seminars in arthritis and rheumatism.

[16]  F. Noyes,et al.  Meniscal material properties are minimally affected by matrix stabilization using glutaraldehyde and glycation with ribose , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[17]  J. F. Woessner,et al.  The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. , 1961, Archives of biochemistry and biophysics.

[18]  K. Athanasiou,et al.  Extraction techniques for the decellularization of tissue engineered articular cartilage constructs. , 2009, Biomaterials.

[19]  Gerard A Ateshian,et al.  Synergistic action of growth factors and dynamic loading for articular cartilage tissue engineering. , 2003, Tissue engineering.

[20]  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.

[21]  J. Fisher,et al.  Development and characterization of an acellular porcine medial meniscus for use in tissue engineering. , 2008, Tissue engineering. Part A.

[22]  J. Katz,et al.  Lumbar disc disorders and low-back pain: socioeconomic factors and consequences. , 2006, The Journal of bone and joint surgery. American volume.

[23]  A. Reddi,et al.  Increased accumulation of superficial zone protein (SZP) in articular cartilage in response to bone morphogenetic protein‐7 and growth factors , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[24]  F. J. Dzida,et al.  Biomechanical Properties of Hip Cartilage in Experimental Animal Models , 1995, Clinical orthopaedics and related research.

[25]  Kyriacos A Athanasiou,et al.  Assessment of a bovine co-culture, scaffold-free method for growing meniscus-shaped constructs. , 2007, Tissue engineering.

[26]  P. Pirttiniemi,et al.  Comparison of Amounts and Properties of Collagen and Proteoglycans in Condylar, Costal and Nasal Cartilages , 1999, Cells Tissues Organs.

[27]  K. Minakuchi,et al.  Chondrocyte migration to fibronectin, type I collagen, and type II collagen. , 1997, Cell structure and function.

[28]  M. Sandrin,et al.  Galα(1,3)Gal, the Major Xenoantigen(s) Recognised in Pigs by Human Natural Antibodies , 1994, Immunological reviews.

[29]  D. Mooney,et al.  Combining chondrocytes and smooth muscle cells to engineer hybrid soft tissue constructs. , 2000, Tissue engineering.

[30]  Kyriacos A Athanasiou,et al.  Success rates and immunologic responses of autogenic, allogenic, and xenogenic treatments to repair articular cartilage defects. , 2009, Tissue engineering. Part B, Reviews.

[31]  I. N. Sneddon The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile , 1965 .

[32]  P. McFetridge,et al.  A mechanical evaluation of three decellularization methods in the design of a xenogeneic scaffold for tissue engineering the temporomandibular joint disc. , 2008, Acta biomaterialia.

[33]  Yu-Ting Tsai,et al.  Process development of an acellular dermal matrix (ADM) for biomedical applications. , 2004, Biomaterials.

[34]  P. Gratzer,et al.  Effectiveness of three extraction techniques in the development of a decellularized bone-anterior cruciate ligament-bone graft. , 2005, Biomaterials.

[35]  Stephen F Badylak,et al.  Quantification of DNA in biologic scaffold materials. , 2009, The Journal of surgical research.