Autologous tissue-engineered trachea with sheep nasal chondrocytes

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Autologous tissue-engineered trachea with sheep nasal chondrocytes Koji Kojima, MD, PhD, Lawrence J. Bonassar, PhD, Amit K. Roy, PhD, Charles A. Vacanti, MD, Joaquin Cortiella, MD The Journal of Thoracic and Cardiovascular Surgery Volume 123, Issue 6, Pages 1177-1184 (June 2002) DOI: 10.1067/mtc.2002.121161 Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Schematic diagram of methods for isolation, culture, and implantation. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Phase-contrast photomicrograph of sheep nasal chondrocytes in monolayer culture (A). (Original magnification 200×.) Nonwoven PGA mesh used for chondrocyte and fibroblast growth (B). Chondrocytes attaching to PGA on day 0 (C) and growth of cells and matrix on day 7 (D). The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 A, Helical template fabricated with a silicone mold-making kit. B, The chondrocyte-seeded matrix was placed in the grooves of the template (arrow), and the entire template was wrapped with the fibroblast-seeded mesh. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 Implantation of TETs in sheep. A, A pocket was created under the sternocleidomastoid muscle, and the implant was placed in this pocket. B, The autologous TET was harvested from the sheep neck at 8 weeks. C, A 5-cm defect in the cervical trachea was created, and the lungs were ventilated with a second endotracheal tube inserted through the operative field. D, The sheep TET was implanted by means of an end-to-end anastomosis. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 5 Lateral view (A) and frontal view (B) of a TET harvested from a nude rat at 8 weeks. In both views similarity to native tracheal tissue is evident. There is a distinct structure containing both cartilage and connective tissue, just as in the normal trachea. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 6 H&E staining: A, rat TET; B, sheep TET; C, native trachea. (Original magnification 100×). Safranin-O stains were deeply positive and indicative of abundant proteoglycan production in each group: D, rat TET; E, sheep TET; F, native trachea. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 7 GAG content of native and engineered tissue (data represent n = 6 ± SD). Statistical difference was determined by means of 1-way analysis of variance. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 8 Hydroxyproline content of native and engineered tissue (data represent n = 6 ± SD). Data were not statistically different by means of 1-way analysis of variance. NS, Not significant. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 9 Cell density of native and engineered tissue (data represent n = 6 ± SD). Statistical difference was determined by means of 1-way analysis of variance. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 1177-1184DOI: (10.1067/mtc.2002.121161) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions