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Development of Acellular Respiratory Mucosal Matrix Using Porcine Tracheal Mucosa

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Respiratory mucosa defects result in airway obstruction and infection, requiring subsequent functional recovery of the respiratory epithelium. Because site-specific extracellular matrix (ECM) facilitates restoration of organ function by promoting cellular migration and engraftment, previous studies considered decellularized trachea an ideal ECM; however, incomplete cell removal from cartilage and mucosal-architecture destruction are frequently reported. Here, we developed a decellularization protocol and applied it to the respiratory mucosa of separated porcine tracheas.

Methods:

The trachea was divided into groups according to decellularization protocol: native mucosa, freezing–thawing (FT), FT followed by the use of Perasafe-based chemical agents before mucosal separation (wFTP), after mucosal separation (mFTP), and followed by DNase decellularization (mFTD). Decellularization efficacy was evaluated by DNA quantification and hematoxylin and eosin staining, and ECM content of the scaffold was evaluated by histologic analysis and glycosaminoglycan and collagen assays. Biocompatibility was assessed by cell-viability assay and in vivo transplantation.

Results:

The mFTP mucosa showed low antigenicity and maintained the ECM to form a proper microstructure. Additionally, tonsil-derived stem cells remained viable when cultured with or seeded onto mFTP mucosa, and the in vivo host response showed a constructive pattern following implantation of the mFTP scaffolds.

Conclusion:

These results demonstrated that xenogenic acellular respiratory mucosa matrix displayed suitable biocompatibility as a scaffold material for respiratory mucosa engineering.

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References

  1. Tsukada H, Osada H. Experimental study of a new tracheal prosthesis: pored Dacron tube. J Thorac Cardiovasc Surg. 2004;127:877–84.

    Article  Google Scholar 

  2. Kim HS, Suh H, Lee JH, Kim JH, Song DE, Jo I, et al. Development of an artificial tracheal prosthesis: a semicircular shape polyurethane scaffold. Tissue Eng Regen Med. 2011;8:439–45.

    Google Scholar 

  3. Chang JW, Park SA, Park JK, Choi JW, Kim YS, Shin YS, et al. Tissue-engineered tracheal reconstruction using three-dimensionally printed artificial tracheal graft: preliminary report. Artif Organs. 2014;38:E95–105.

    Article  CAS  Google Scholar 

  4. Hong HJ, Chang JW, Park JK, Choi JW, Kim YS, Shin YS, et al. Tracheal reconstruction using chondrocytes seeded on a poly(l-lactic-co-glycolic acid)-fibrin/hyaluronan. J Biomed Mater Res A. 2014;102:4142–50.

    Article  Google Scholar 

  5. Lee JH, Park HS, Oh SH, Lee JH, Kim JR, Kim HJ, et al. Triple-layered polyurethane prosthesis with wrinkles for repairing partial tracheal defects. Laryngoscope. 2014;124:2757–63.

    Article  CAS  Google Scholar 

  6. Melgoza EL, Vallicrosa G, Serenó L, Ciurana J, Rodríguez AC. Rapid tooling using 3D printing system for manufacturing of customized tracheal stent. Rapid Prototyp J. 2014;20:2–12.

    Article  Google Scholar 

  7. Shin YS, Lee BH, Choi JW, Min BH, Chang JW, Yang SS, et al. Tissue-engineered tracheal reconstruction using chondrocyte seeded on a porcine cartilage-derived substance scaffold. Int J Pediatr Otorhinolaryngol. 2014;78:32–8.

    Article  Google Scholar 

  8. Chua M, Chui CK, Teo C, Lau D. Patient-specific carbon nanocomposite tracheal prosthesis. Int J Artif Organs. 2015;38:31–8.

    Article  Google Scholar 

  9. Park JH, Hong JM, Ju YM, Jung JW, Kang HW, Lee SJ, et al. A novel tissue-engineered trachea with a mechanical behavior similar to native trachea. Biomaterials. 2015;62:106–15.

    Article  CAS  Google Scholar 

  10. Shin YS, Choi JW, Park JK, Kim YS, Yang SS, Min BH, et al. Tissue-engineered tracheal reconstruction using mesenchymal stem cells seeded on a porcine cartilage powder scaffold. Ann Biomed Eng. 2015;43:1003–13.

    Article  Google Scholar 

  11. Jung SY, Lee SJ, Kim HY, Park HS, Wang Z, Kim HJ, et al. 3D printed polyurethane prosthesis for partial tracheal reconstruction: a pilot animal study. Biofabrication. 2016;8:045015.

    Article  Google Scholar 

  12. O’Leary C, Cavanagh B, Unger RE, Kirkpatrick CJ, O’Dea S, O’Brien FJ, et al. The development of a tissue-engineered tracheobronchial epithelial model using a bilayered collagen-hyaluronate scaffold. Biomaterials. 2016;85:111–27.

    Article  Google Scholar 

  13. Butler CR, Hynds RE, Crowley C, Gowers KH, Partington L, Hamilton NJ, et al. Vacuum-assisted decellularization: an accelerated protocol to generate tissue-engineered human tracheal scaffolds. Biomaterials. 2017;124:95–105.

    Article  CAS  Google Scholar 

  14. Baiguera S, Jungebluth P, Burns A, Mavilia C, Haag J, De Coppi P, et al. Tissue engineered human tracheas for in vivo implantation. Biomaterials. 2010;31:8931–8.

    Article  CAS  Google Scholar 

  15. Partington L, Mordan NJ, Mason C, Knowles JC, Kim HW, Lowdell MW, et al. Biochemical changes caused by decellularization may compromise mechanical integrity of tracheal scaffolds. Acta Biomater. 2013;9:5251–61.

    Article  CAS  Google Scholar 

  16. Zang M, Zhang Q, Chang EI, Mathur AB, Yu P. Decellularized tracheal matrix scaffold for tracheal tissue engineering: in vivo host response. Plast Reconstr Surg. 2013;132:549e–59.

    Article  CAS  Google Scholar 

  17. Sun F, Pan S, Shi HC, Zhang FB, Zhang WD, Ye G, et al. Structural integrity, immunogenicity and biomechanical evaluation of rabbit decelluarized tracheal matrix. J Biomed Mater Res A. 2015;103:1509–19.

    Article  Google Scholar 

  18. Den Hondt M, Vanaudenaerde BM, Maughan EF, Butler CR, Crowley C, Verbeken EK, et al. An optimized non-destructive protocol for testing mechanical properties in decellularized rabbit trachea. Acta Biomater. 2017;60:291–301.

    Article  Google Scholar 

  19. Xu Y, Li D, Yin Z, He A, Lin M, Jiang G, et al. Tissue-engineered trachea regeneration using decellularized trachea matrix treated with laser micropore technique. Acta Biomater. 2017;58:113–21.

    Article  CAS  Google Scholar 

  20. Park JH, Jung JW, Kang HW, Joo YH, Lee JS, Cho DW. Development of a 3D bellows tracheal graft: mechanical behavior analysis, fabrication and an in vivo feasibility study. Biofabrication. 2012;4:035004.

    Article  Google Scholar 

  21. Chua CHM, Chui CK, Rai B, Lau DPD. Development of a patient specific artificial tracheal prosthesis: design, mechanical behavior analysis and manufacturing. Conf Proc IEEE Eng Med Biol Soc. 2013;2013:6236–9.

    Google Scholar 

  22. Macchiarini P, Jungebluth P, Go T, Asnaghi MA, Rees LE, Cogan TA, et al. Clinical transplantation of a tissue-engineered airway. Lancet. 2008;372:2023–30.

    Article  Google Scholar 

  23. Delaere P, Vranckx J, Verleden G, De Leyn P, Van Raemdonck D, Leuven Tracheal Transplant Group. Tracheal allotransplantation after withdrawal of immunosuppressive therapy. N Engl J Med. 2010;362:138–45.

    Article  CAS  Google Scholar 

  24. Gui L, Chan SA, Breuer CK, Niklason LE. Novel utilization of serum in tissue decellularization. Tissue Eng Part C Methods. 2010;16:173–84.

    Article  CAS  Google Scholar 

  25. Hussein KH, Park KM, Kang KS, Woo HM. Biocompatibility evaluation of tissue-engineered decellularized scaffolds for biomedical application. Mater Sci Eng C Mater Biol Appl. 2016;67:766–78.

    Article  CAS  Google Scholar 

  26. Xu S, Lu F, Cheng L, Li C, Zhou X, Wu Y, et al. Preparation and characterization of small-diameter decellularization scaffolds for vascular tissue engineering in an animal model. Biomed Eng Online. 2017;16:55.

    Article  Google Scholar 

  27. Keane TJ, Londono R, Carey RM, Carruthers CA, Reing JE, Dearth CL, et al. Preparation and characterization of a biologic scaffold from esophageal mucosa. Biomaterials. 2013;34:6729–37.

    Article  CAS  Google Scholar 

  28. Valentin JE, Badylak JS, McCabe GP, Badylak SF. Extracellular matrix bioscaffolds for orthopaedic applications: a comparative histologic study. J Bone Joint Surg Am. 2006;88:2673–86.

    Article  Google Scholar 

  29. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials. 2011;32:32–43.

    Article  Google Scholar 

  30. Allen RA, Seltz LM, Jiang H, Kasick RT, Sellaro TL, Badylak SF, et al. Adrenal extracellular matrix scaffolds support adrenocortical cell proliferation and function in vitro. Tissue Eng Part A. 2010;16:3363–74.

    Article  CAS  Google Scholar 

  31. French KM, Boopathy AV, DeQuach JA, Chingozha L, Lu H, Christman KL, et al. A naturally derived cardiac extracellular matrix enhances cardiac progenitor cell behavior in vitro. Acta Biomater. 2012;8:4357–64.

    Article  CAS  Google Scholar 

  32. Sellaro TL, Ravindra AK, Stolz DB, Badylak SF. Maintenance of hepatic sinusoidal endothelial cell phenotype in vitro using organ-specific extracellular matrix scaffolds. Tissue Eng. 2007;13:2301–10.

    Article  CAS  Google Scholar 

  33. Morris AH, Chang J, Kyriakides TR. Inadequate processing of decellularized dermal matrix reduces cell viability in vitro and increases apoptosis and acute inflammation in vivo. Biores Open Access. 2016;5:177–87.

    Article  CAS  Google Scholar 

  34. Wong ML, Griffiths LG. Immunogenicity in xenogeneic scaffold generation: antigen removal vs. decellularization. Acta Biomater. 2014;10:1806–16.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by 2016 Young Medical Science Researcher Grants from Ewha Womans University College of Medicine and grants from the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology (Grant No. NRF-2017R1D1A1B03034399).

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Correspondence to Han Su Kim.

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The authors have no conflicts of interest to disclose.

Ethical statement

The animal studies were performed after receiving approval of the Institutional Animal Care and Use Committee in Ewha Womans University Medical Research Institute (Approval No. ESM-.18-0404).

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Jung, S.Y., Tran, A.NT., Kim, H.Y. et al. Development of Acellular Respiratory Mucosal Matrix Using Porcine Tracheal Mucosa. Tissue Eng Regen Med 17, 433–443 (2020). https://doi.org/10.1007/s13770-020-00260-w

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  • DOI: https://doi.org/10.1007/s13770-020-00260-w

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