Long-term outcomes of patch tracheoplasty using collagenous tissue membranes (biosheets) produced by in-body tissue architecture in a beagle model

  • Satoshi Umeda
  • Yasuhide Nakayama
  • Takeshi Terazawa
  • Ryosuke Iwai
  • Shohei Hiwatashi
  • Kengo Nakahata
  • Yuichi Takama
  • Hiroomi OkuyamaEmail author
Original Article



Although various artificial tracheas have been developed, none have proven satisfactory for clinical use. In-body tissue architecture (IBTA) has enabled us to produce collagenous tissues with a wide range of shapes and sizes to meet the needs of individual recipients. In the present study, we investigated the long-term outcomes of patch tracheoplasty using an IBTA-induced collagenous tissue membrane (“biosheet”) in a beagle model.


Nine adult female beagles were used. Biosheets were prepared by embedding cylindrical molds assembled with a silicone rod and a slitting pipe into dorsal subcutaneous pouches for 2 months. The sheets were then implanted by patch tracheoplasty. An endoscopic evaluation was performed after 1, 3, or 12 months. The implanted biosheets were harvested for a histological evaluation at the same time points.


All animals survived the study. At 1 month after tracheoplasty, the anastomotic parts and internal surface of the biosheets were smooth with ciliated columnar epithelium, which regenerated into the internal surface of the biosheet. The chronological spread of chondrocytes into the biosheet was observed at 3 and 12 months.


Biosheets showed excellent performance as a scaffold for trachea regeneration with complete luminal epithelium and partial chondrocytes in a 1-year beagle implantation model of patch tracheoplasty.


Regenerative medicine Trachea Scaffold Patch tracheoplasty Dog (beagle) 


Compliance with ethical standards

Conflict of interest

All authors declare no conflict of interest.


  1. 1.
    Ott LM, Vu CH, Farris AL, Fox KD, Galbraith RA, Weiss ML, et al. Functional reconstruction of tracheal defects by protein-loaded, cell-seeded, fibrous constructs in rabbits. Tissue Eng Part A. 2015;21:2390–403.CrossRefGoogle Scholar
  2. 2.
    Nakayama Y, Ishibashi-Ueda H, Takamizawa K. In vivo tissue-engineered small-caliber arterial graft prosthesis consisting of autologous tissue (biotube). Cell Transpl. 2004;13:439–49.CrossRefGoogle Scholar
  3. 3.
    Yamanami M, Ishibashi-Ueda H, Yamamoto A, Iida H, Watanabe T, Kanda K, et al. Transplantation study of small-caliber ‘‘biotube’’ vascular grafts in a rat model. J Artif Organs. 2013;16:59–655.CrossRefGoogle Scholar
  4. 4.
    Watanabe T, Kanda K, Ishibashi-Ueda H, Yaku H, Nakayama Y. Autologous small-caliber “biotube” vascular grafts with argatroban loading: a histomorphological examination after transplantation to rabbits. J Biomed Mater Res B Appl Biomater. 2010;92:236–42.CrossRefGoogle Scholar
  5. 5.
    Okuyama H, Umeda S, Takama Y, Terasawa T, Nakayama Y. Patch esophagoplasty using an in-body-tissue-engineered collagenous connective tissue membrane. J Pediatr Surg. 2017;53:223–6.CrossRefGoogle Scholar
  6. 6.
    Satake R, Komura M, Komura H, Kodaka T, Terawaki K, Ikebukuro K, et al. Patch tracheoplasty in body tissue engineering using collagenous connective tissue membranes (biosheets). J Pediatr Surg. 2013;51:244–8.CrossRefGoogle Scholar
  7. 7.
    Jungebluth P, Haag JC, Sjöqvist S, Gustafsson Y, Beltrán Rodríguez A, Del Gaudio C, et al. Tracheal tissue engineering in rats. Nat Protoc. 2014;9:2164–79.CrossRefGoogle Scholar
  8. 8.
    Delaere PR, Raemdonck DV. The trachea: the first tissue-engineered organ? J Thorac Cardiovasc Surg. 2014;147:1128–32.CrossRefGoogle Scholar
  9. 9.
    Grillo HC. Tracheal replacement: a critical review. Ann Thorac Surg. 2002;73:1995–2004.CrossRefGoogle Scholar
  10. 10.
    Tsugawa C, Nishijima E, Muraji T, Satoh S, Takamizawa S, Yamaguchi M, et al. Tracheoplasty for long segment congenital tracheal stenosis: analysis of 29 patients over two decades. J Pediatr Surg. 2003;38:1703–6.CrossRefGoogle Scholar
  11. 11.
    Oue T, Kamata S, Usui N, Okuyama H, Nose K, Okada A. Histopathologic changes after tracheobronchial reconstruction with costal cartilage graft for congenital tracheal stenosis. J Pediatr Surg. 2001;36:329–33.CrossRefGoogle Scholar
  12. 12.
    Yazdanbakhsh AP, van Rijssen LB, Koolbergen DR, König A, de Mol BA, Hazekamp MG. Long-term follow-up of tracheoplasty using autologous pericardial patch and strips of costal cartilage. Eur J Cardiothorac Surg. 2015;47:146–52.CrossRefGoogle Scholar
  13. 13.
    Fanous N, Husain SA, Ruzmetov M, Rodefeld MD, Turrentine MW, Brown JW. Anterior pericardial tracheoplasty for long-segment tracheal stenosis: long-term outcomes. J Thorac Cardiovasc Surg. 2010;139:18–23.CrossRefGoogle Scholar
  14. 14.
    Hayashida K, Kanda K, Yaku H, Ando J, Nakayama Y. Development of an in vivo tissue-engineered, autologous heart valve (the biovalve): preparation of a prototype model. J Thorac Cardiovasc Surg. 2007;134:152–9.CrossRefGoogle Scholar
  15. 15.
    Takiyama N, Mizuno T, Iwai R, Uechi M, Nakayama Y. In-body tissue-engineered collagenous connective tissue membranes (BIOSHEETs) for potential corneal stromal substitution. J Tissue Eng Regen Med. 2016;10:E518–26.CrossRefGoogle Scholar
  16. 16.
    Yamanami M, Yahata Y, Uechi M, Fujiwara M, Ishibashi-Ueda H, Kanda K, et al. Development of a completely autologous valved conduit with the sinus of Valsalva using in-body tissue architecture technology: a pilot study in pulmonary valve replacement in a beagle model. Circulation. 2010;122:S100–6.CrossRefGoogle Scholar
  17. 17.
    Nakayama Y, Oshima N, Tatsumi E, Ichii O, Nishimura T. iBTA-induced bovine Biosheet for repair of abdominal wall defects in a beagle model: proof of concept. Hernia. 2018;

Copyright information

© Springer Nature Singapore Pte Ltd. 2019

Authors and Affiliations

  • Satoshi Umeda
    • 1
  • Yasuhide Nakayama
    • 2
  • Takeshi Terazawa
    • 3
  • Ryosuke Iwai
    • 4
  • Shohei Hiwatashi
    • 1
  • Kengo Nakahata
    • 1
  • Yuichi Takama
    • 1
  • Hiroomi Okuyama
    • 1
    Email author
  1. 1.Department of Pediatric SurgeryOsaka University Graduate School of MedicineOsakaJapan
  2. 2.Biotube Co., Ltd.TokyoJapan
  3. 3.Division of Cell Engineering, Graduate School of Chemical Science and EngineeringHokkaido UniversitySapporoJapan
  4. 4.Research Institute of Technology, Okayama University of ScienceOkayamaJapan

Personalised recommendations