Biomimetic design and fabrication of porous chitosan–gelatin liver scaffolds with hierarchical channel network

  • Haibo Gong
  • Jephte Agustin
  • David Wootton
  • Jack G. Zhou


The presence of a hierarchical channel network in tissue engineering scaffold is essential to construct metabolically demanding liver tissue with thick and complex structures. In this research, chitosan–gelatin (C/G) scaffolds with fine three-dimensional channels were fabricated using indirect solid freeform fabrication and freeze-drying techniques. Fabrication processes were studied to create predesigned hierarchical channel network inside C/G scaffolds and achieve desired porous structure. Static in-vitro cell culture test showed that HepG2 cells attached on both micro-pores and micro-channels in C/G scaffolds successfully. HepG2 proliferated at much higher rates on C/G scaffolds with channel network, compared with those without channels. This approach demonstrated a promising way to engineer liver scaffolds with hierarchical channel network, and may lead to the development of thick and complex liver tissue equivalent in the future.


Chitosan HepG2 Cell Channel Network Porous Scaffold Genipin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We gratefully thank National Science Foundation (NSF) for the support DMI-0300405, CMMI-0700139 and CMMI-0925348, to let us to conduct this challenging project. We are grateful that Professor Peter I. Lelkes generously provided cell culture related facility. We also would like to thank Dr. Qingwei Zhang for the help in SEM, Dr. Jingjia Han for the help in cell culture, Pimchanok Pimton for the help in confocal microscope, Dolores Conover for the help in freeze-drying. The Centralized Research Facility (CRF) of the College of Engineering at Drexel University provided access to electronic microscopes used in this work.


  1. 1.
    Liu Tsang V, Bhatia SN. Three-dimensional tissue fabrication. Adv Drug Deliv Rev. 2004;56(11):1635–47.CrossRefGoogle Scholar
  2. 2.
    Yeong W-Y, et al. Rapid prototyping in tissue engineering: challenges and potential. Trends Biotechnol. 2004;22(12):643–52.CrossRefGoogle Scholar
  3. 3.
    Mikos AG, et al. Preparation and characterization of poly(l-lactic acid) foams. Polymer. 1994;35(5):1068–77.CrossRefGoogle Scholar
  4. 4.
    Kim U-J, et al. Three-dimensional aqueous-derived biomaterial scaffolds from silk fibroin. Biomaterials. 2005;26(15):2775–85.CrossRefGoogle Scholar
  5. 5.
    Kang H-W, Tabata Y, Ikada Y. Fabrication of porous gelatin scaffolds for tissue engineering. Biomaterials. 1999;20(14):1339–44.CrossRefGoogle Scholar
  6. 6.
    Madihally SV, Matthew HWT. Porous chitosan scaffolds for tissue engineering. Biomaterials. 1999;20(12):1133–42.CrossRefGoogle Scholar
  7. 7.
    Mao JS, et al. Structure and properties of bilayer chitosan–gelatin scaffolds. Biomaterials. 2003;24(6):1067–74.CrossRefGoogle Scholar
  8. 8.
    Radisic M, et al. Medium perfusion enables engineering of compact and contractile cardiac tissue. Am J Physiol Heart Circ Physiol. 2004;286(2):H507–16.CrossRefGoogle Scholar
  9. 9.
    Miller JS, et al. Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. Nat Mater. 2012;11(9):768–74.CrossRefGoogle Scholar
  10. 10.
    He J, et al. Indirect fabrication of microstructured chitosan–gelatin scaffolds using rapid prototyping. Virtual Phys Prototyp. 2008;3(3):159–66.CrossRefGoogle Scholar
  11. 11.
    Jiankang H, et al. Fabrication and characterization of chitosan/gelatin porous scaffolds with predefined internal microstructures. Polymer. 2007;48(15):4578–88.CrossRefGoogle Scholar
  12. 12.
    Li K, et al. Chitosan/gelatin composite microcarrier for hepatocyte culture. Biotechnol Lett. 2004;26(11):879–83.CrossRefGoogle Scholar
  13. 13.
    Huang Y, et al. In vitro characterization of chitosan–gelatin scaffolds for tissue engineering. Biomaterials. 2005;26(36):7616–27.CrossRefGoogle Scholar
  14. 14.
    Nikolaychik VV, Samet MM, Lelkes PI. A new method for continual quantitation of viable cells on endothelialized polyurethanes. J Biomater Sci Polym Ed. 1996;7:881–91.CrossRefGoogle Scholar
  15. 15.
    Huang J-H, et al. Rapid Fabrication of Bio-inspired 3D Microfluidic Vascular Networks. Adv Mater. 2009;21(35):3567–71.CrossRefGoogle Scholar
  16. 16.
    Wu Willie CJH, Aragón AM. Direct-write assembly of biomimetic microvascular networks for efficient fluid transport. Soft Matter. 2010;6:739–42.CrossRefGoogle Scholar
  17. 17.
    Sakai Y, et al. Toward engineering of vascularized three-dimensional liver tissue equivalents possessing a clinically significant mass. Biochem Eng J. 2010;48(3):348–61.CrossRefGoogle Scholar
  18. 18.
    Hoganson DMP, Howard IAU, Vacanti JP. Tissue engineering and organ structure: a vascularized approach to liver and lung. Pediatr Res. 2008;63(5):520–6.CrossRefGoogle Scholar
  19. 19.
    Kaihara S, Koka JBR, Ochoa ER, Ravens M, Pien H, Cunningham B, Vacanti JP. Silicon micromachining to tissue engineer branched vascular channels for liver fabrication. Tissue Eng. 2000;6:105–17.CrossRefGoogle Scholar
  20. 20.
    Lee M, Wu BM, Dunn JCY. Effect of scaffold architecture and pore size on smooth muscle cell growth. J Biomed Mater Res Part A. 2008;87A(4):1010–6.CrossRefGoogle Scholar
  21. 21.
    Ranucci CS, Moghe PV. Polymer substrate topography actively regulates the multicellular organization and liver-specific functions of cultured hepatocytes. Tissue Eng. 1999;5(5):407–20.CrossRefGoogle Scholar
  22. 22.
    Ranucci CS, et al. Control of hepatocyte function on collagen foams: sizing matrix pores toward selective induction of 2-D and 3-D cellular morphogenesis. Biomaterials. 2000;21(8):783–93.CrossRefGoogle Scholar
  23. 23.
    Kim SS, Utsunomiya H, Koski JA, Wu BM, Cima MJ, Sohn J, Mukai K, Griffith LG, Vacanti JP. Survival and function of hepatocytes on a novel three-dimensional synthetic biodegradable polymer scaffold with an intrinsic network of channels. Ann. Surg. 1998;228:8–13.CrossRefGoogle Scholar
  24. 24.
    Robert Lanza, R.L., Principle of Tissue Engineering. 2007: Academic Press.Google Scholar
  25. 25.
    Yang S, et al. The Design of Scaffolds for Use in Tissue Engineering Part I. Tradit Factors Tissue Eng. 2001;7(6):679–89.CrossRefGoogle Scholar
  26. 26.
    Sakai Y. A novel poly–lactic acid scaffold that possesses a macroporous structure and a branching/joining three-dimensional flow channel network: its fabrication and application to perfusion culture of human hepatoma Hep G2 cells. Mater Sci Eng C. 2004;24(3):379–86.CrossRefGoogle Scholar
  27. 27.
    Huang H, et al. Avidin–biotin binding-based cell seeding and perfusion culture of liver-derived cells in a porous scaffold with a three-dimensional interconnected flow-channel network. Biomaterials. 2007;28(26):3815–23.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Haibo Gong
    • 1
  • Jephte Agustin
    • 1
  • David Wootton
    • 2
  • Jack G. Zhou
    • 1
  1. 1.Department of Mechanical Engineering and MechanicsDrexel UniversityPhiladelphiaUSA
  2. 2.Department of Mechanical EngineeringCooper UnionNew YorkUSA

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