A composite tissue‐engineered trachea using sheep nasal chondrocyte and epithelial cells

This study evaluates the feasibility of producing a composite engineered tracheal equivalent composed of cylindrical cartilaginous structures with lumens lined with nasal epithelial cells. Chondrocytes and epithelial cells isolated from sheep nasal septum were cultured in Ham's F12 media. After 2 wk, chondrocyte suspensions were seeded onto a matrix of polyglycolic acid. Cell‐polymer constructs were wrapped around silicon tubes and cultured in vitro for 1 wk, followed by implanting into subcutaneous pockets on the backs of nude mice. After 6 wk, epithelial cells were suspended in a hydrogel and injected into the embedded cartilaginous cylinders following removal of the silicon tube. Implants were harvested 4 wk later and analyzed. The morphology of implants resembles that of native sheep trachea. H&E staining shows the presence of mature cartilage and formation of a pseudostratified columnar epithelium, with a distinct interface between tissue‐engineered cartilage and epithelium. Safranin‐O staining shows that tissue‐engineered cartilage is organized into lobules with round, angular lacunae, each containing a single chondrocyte. Proteoglycan and hydroxyproline contents are similar to native cartilage. This study demonstrates the feasibility of recreating the cartilage and epithelial portion of the trachea using tissue harvested in a single procedure. This has the potential to facilitate an autologous repair of segmental tracheal defects.—Kojima, K., Bonassar, L. J., Roy, A. K., Mizuno, H., Cortiella, J., Vacanti, C. A. A composite tissue‐engineered trachea using sheep nasal chondrocyte and epithelial cells. FASEB J. 17, 823–828 (2003)

[1]  L. Bonassar,et al.  Age-related changes in the composition and mechanical properties of human nasal cartilage. , 2002, Archives of biochemistry and biophysics.

[2]  A. Cole,et al.  Comparison of biomechanical and biochemical properties of cartilage from human knee and ankle pairs , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[3]  M. Aoe,et al.  Origin of regenerated epithelium in cryopreserved tracheal allotransplantation. , 1998, The Annals of thoracic surgery.

[4]  Charles A Vacanti,et al.  Age dependence of biochemical and biomechanical properties of tissue-engineered human septal cartilage. , 2002, Biomaterials.

[5]  J. Vacanti,et al.  Bone and cartilage reconstruction with tissue engineering approaches. , 1994, Otolaryngologic clinics of North America.

[6]  L. Bonassar,et al.  Age dependence of cellular properties of human septal cartilage: implications for tissue engineering. , 2001, Archives of otolaryngology--head & neck surgery.

[7]  T. Mitsudomi,et al.  Maximum length of tracheal autografts in dogs. , 1993, The Journal of thoracic and cardiovascular surgery.

[8]  J. Vacanti,et al.  Experimental tracheal replacement using tissue-engineered cartilage. , 1994, Journal of pediatric surgery.

[9]  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.

[10]  J. Barker,et al.  Prefabricated Jejunal Free‐Tissue Transfer for Tracheal Reconstruction: An Experimental Study , 1996, Plastic and reconstructive surgery.

[11]  Neville We Reconstruction of the trachea and stem bronchi with Neville prosthesis. , 1982 .

[12]  H. Stegemann,et al.  Determination of hydroxyproline. , 1967, Clinica chimica acta; international journal of clinical chemistry.

[13]  I. Kessel,et al.  Congenital Absence of the Trachea , 1953, Thorax.

[14]  D L Bader,et al.  Quantification of sulfated glycosaminoglycans in chondrocyte/alginate cultures, by use of 1,9-dimethylmethylene blue. , 1996, Analytical biochemistry.

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

[16]  Y Ikada,et al.  The repair of tracheal defects using bioabsorbable mesh. , 1992, ASAIO journal.

[17]  M. Onoe,et al.  The first step of experimental study on hybrid trachea: use of cultured fibroblasts with artificial matrix. , 1994 .

[18]  P. Cavadas Tracheal Reconstruction Using a Free Jejunal Flap with Cartilage Skeleton: Experimental Study , 1998, Plastic and reconstructive surgery.