Tissue engineering of an auricular cartilage model utilizing cultured chondrocyte-poly(L-lactide-epsilon-caprolactone) scaffolds.

To determine the potential development in vivo of tissue-engineered auricular cartilage, chondrocytes from articular cartilage of bovine forelimb joints were seeded on poly(L-lactic acid-epsilon-caprolactone) copolymer scaffolds molded into the shape of a human ear. Copolymer scaffolds alone in the same shape were studied for comparison. Chondrocyte-seeded copolymer constructs and scaffolds alone were each implanted in dorsal skin flaps of athymic mice for up to 40 weeks. Retrieved specimens were examined by histological and molecular techniques. After 10 weeks of implantation, cell-seeded constructs developed cartilage as assessed by toluidine blue and safranin-O red staining; a vascular, perichondrium-like capsule enveloped these constructs; and tissue formation resembled the auricular shape molded originally. Cartilage matrix formation increased, the capsule persisted, and initial auricular configuration was maintained through implantation for 40 weeks. The presence of cartilage production was correlated with RT-PCR analysis, which showed expression of bovine-specific type II collagen and aggrecan mRNA in cell-seeded specimens at 20 and 40 weeks. Copolymer scaffolds monitored only for 40 weeks failed to develop cartilage or a defined capsule and expressed no mRNA. Extensive vascularization led to scaffold erosion, decrease in original size, and loss of contour and shape. These results demonstrate that poly(L-lactic acid-epsilon-caprolactone) copolymer seeded with articular chondrocytes supports development and maintenance of cartilage in a human ear shape over periods to 40 weeks in this implantation model.

[1]  A. Atala,et al.  Bladder augmentation using allogenic bladder submucosa seeded with cells. , 1998, Urology.

[2]  B. Obradovic,et al.  Integration of engineered cartilage , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[3]  G. Naughton,et al.  Evaluation of matrix scaffolds for tissue engineering of articular cartilage grafts. , 1997, Journal of biomedical materials research.

[4]  V. R. Patel,et al.  Salvage of the head of the radius after fracture-dislocation of the elbow , 1999 .

[5]  W M Lai,et al.  Fluid transport and mechanical properties of articular cartilage: a review. , 1984, Journal of biomechanics.

[6]  E B Hunziker,et al.  Articular cartilage repair: are the intrinsic biological constraints undermining this process insuperable? , 1999, Osteoarthritis and cartilage.

[7]  C. Vacanti,et al.  Tissue-engineered morphogenesis of cartilage and bone by means of cell transplantation using synthetic biodegradable polymer matrices. , 1994, Clinics in plastic surgery.

[8]  D. Amiel,et al.  In situ assessment of cell viability within biodegradable polylactic acid polymer matrices. , 1995, Biomaterials.

[9]  R. Giardino,et al.  Preparation of a new nerve guide from a poly(L-lactide-co-6-caprolactone). , 1994, Biomaterials.

[10]  N. Parenteau,et al.  Development of a bilayered living skin construct for clinical applications , 1994, Biotechnology and bioengineering.

[11]  R Langer,et al.  Creation of viable pulmonary artery autografts through tissue engineering. , 1998, The Journal of thoracic and cardiovascular surgery.

[12]  V. Mow,et al.  Composition and dynamics of articular cartilage: structure, function, and maintaining healthy state. , 1998, The Journal of orthopaedic and sports physical therapy.

[13]  M. Ueda,et al.  Cartilage formation by cultured chondrocytes in a new scaffold made of poly(L-lactide-ϵ-caprolactone) sponge , 2000 .

[14]  S W O'Driscoll,et al.  Role of oxygen tension during cartilage formation by periosteum , 1997, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[15]  N. Hibino,et al.  Tissue-engineered vascular autograft: inferior vena cava replacement in a dog model. , 2001, Tissue engineering.

[16]  R Langer,et al.  Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. , 1993, Journal of biomedical materials research.

[17]  E. Bell,et al.  Strategy for the selection of scaffolds for tissue engineering. , 1995, Tissue engineering.

[18]  V C Mow,et al.  Variations in the intrinsic mechanical properties of human articular cartilage with age, degeneration, and water content. , 1982, The Journal of bone and joint surgery. American volume.

[19]  R C Tanzer,et al.  TOTAL RECONSTRUCTION OF THE AURICLE: The Evolution of a Plan of Treatment , 1971, Plastic and reconstructive surgery.

[20]  A. Schindler,et al.  Sustained drug delivery systems. I. The permeability of poly(ϵ-caprolactone), poly(DL-lactic acid), and their copolymers , 1979 .

[21]  Audrey U. Smith Survival of Frozen Chondrocytes Isolated from Cartilage of Adult Mammals , 1965, Nature.

[22]  J. Buckwalter Articular cartilage: injuries and potential for healing. , 1998, The Journal of orthopaedic and sports physical therapy.

[23]  V C Mow,et al.  Tensile properties of human knee joint cartilage: I. Influence of ionic conditions, weight bearing, and fibrillation on the tensile modulus , 1986, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[24]  T. Matsuda,et al.  Muscular tissue engineering: capillary-incorporated hybrid muscular tissues in vivo tissue culture. , 1998, Cell transplantation.

[25]  W. Bonner,et al.  Isolation and culture of chondrocytes from human adult articular cartilage. , 1967, Arthritis and rheumatism.

[26]  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.

[27]  D E Ingber,et al.  Cellular control lies in the balance of forces. , 1998, Current opinion in cell biology.

[28]  R Langer,et al.  Design of nasoseptal cartilage replacements synthesized from biodegradable polymers and chondrocytes. , 1994, Biomaterials.

[29]  G. Vunjak‐Novakovic,et al.  Gas exchange is essential for bioreactor cultivation of tissue engineered cartilage. , 1999, Biotechnology and bioengineering.

[30]  D. Grijpma,et al.  High molecular weight copolymers of l-lactide and ε-caprolactone as biodegradable elastomeric implant materials , 1991 .

[31]  R Langer,et al.  Development of biomechanical properties and morphogenesis of in vitro tissue engineered cartilage. , 1995, Journal of biomedical materials research.

[32]  Tanzer Rc Microtia--a long-term follow-up of 44 reconstructed auricles. , 1978 .

[33]  C. Wirth,et al.  Techniques of cartilage growth enhancement: a review of the literature. , 1996, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[34]  R. C. Tanzer Editorial: Replantation—problems solved and unsolved , 1967 .

[35]  K. Leong,et al.  The design of scaffolds for use in tissue engineering. Part I. Traditional factors. , 2001, Tissue engineering.

[36]  V. Patel,et al.  Salvage of the head of the radius after fracture-dislocation of the elbow. A case report. , 1999, The Journal of bone and joint surgery. British volume.

[37]  Charles A. Vacanti,et al.  Transplantation of Chondrocytes Utilizing a Polymer‐Cell Construct to Produce Tissue‐Engineered Cartilage in the Shape of a Human Ear , 1997, Plastic and reconstructive surgery.

[38]  M. Klagsbrun Large-scale preparation of chondrocytes. , 1979, Methods in enzymology.

[39]  J. Vacanti,et al.  Tracheal composites tissue engineered from chondrocytes, tracheal epithelial cells, and synthetic degradable scaffolding. , 1994, Transplantation proceedings.

[40]  J. Vacanti,et al.  Synthetic Polymers Seeded with Chondrocytes Provide a Template for New Cartilage Formation , 1991, Plastic and reconstructive surgery.

[41]  G. Kimmel,et al.  Biodegradable Polymers for Sustained Drug Delivery , 1977 .

[42]  I. Arvanitoyannis,et al.  Enzymatic Hydrolysis of Poly (L-lactide-co-ε-caprolactone) s , 1995 .

[43]  C. Patrick,et al.  Preadipocyte seeded PLGA scaffolds for adipose tissue engineering. , 1999, Tissue engineering.

[44]  L. Liotta,et al.  Laser Capture Microdissection , 1996, Science.