Science China Chemistry

, Volume 56, Issue 12, pp 1701–1709 | Cite as

Preparation of enzymatically cross-linked sulfated chitosan hydrogel and its potential application in thick tissue engineering

  • ZhiPing Chen
  • Wei WangEmail author
  • Lei Guo
  • YanYan Yu
  • Zhi YuanEmail author


For the requirement of preliminary vascularization, the scaffolds for thick tissue engineering should possess not only good cell affinity, but also anticoagulant ability. In this paper, an enzymatically crosslinked hydrogel scaffold based on sulfated chitosan (SCTS) was prepared. Firstly, sulfated chitosan-hydroxyphenylpionic acid (SCTS-HPA) conjugate was synthesized, and its structure was identified by FITR and 1H NMR. And then an enzymatically crosslinked hydrogel was prepared in the presence of horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). The gelation time, mechanical property, morphology and cytotoxicity to human umbilical vein endothelial cells (HUVECs) of the hydrogel was evaluated in vitro, the tissue compatibility of SCTS scaffold was studied in vivo. The results showed that the gelation time, mechanical property, morphology of the dehydrated hydrogel could be controlled by the HRP and H2O2 concentration. The cytotoxicity test showed that the hydrogel extracts had no cytotoxicity to HUVECs. The in vivo assay indicated that SCTS-HPA scaffold showed good tissue compatibility, and no thrombus formation. All these results indicated that the SCTS-HPA scaffold could be used as thick tissue engineering scaffold.


enzymatically crosslinked thick tissue engineering scaffold thrombus formation tissue compatibility 


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  1. 1.
    Jain RK, Au P, Tam J, Duda DG, Fukumura D. Engineering vascularized tissue. Nat Biotechnol, 2005, 23: 821–823CrossRefGoogle Scholar
  2. 2.
    Singh DK, Ray AR. Biomedical applications of chitin, chitosan, and their derivatives. J Macromol Sci. Part C: Polymer Reviews, 2000, 40: 69–83CrossRefGoogle Scholar
  3. 3.
    Amiji MM. Surface modification of chitosan membranes by complexation-interpenetration of anionic polysaccharides for improved blood compatibility in hemodialysis. J Biomater Sci Polym Ed, 1996, 8: 281–98CrossRefGoogle Scholar
  4. 4.
    Chen XG, Park HJ. Chemical characteristics of O-carboxymethyl chitosans related to the preparation conditions. Carbohyd Polym, 2003, 53: 355–359CrossRefGoogle Scholar
  5. 5.
    Davis SS, Lin W, Bignotti Fabio, Ferruti P. Conjugate of polyethylene glycol and chitosan. United State Patent, US 6730735 B2, 2004-05-04Google Scholar
  6. 6.
    Jayakumar R, New N, Tokura S, Tamura H. Sulfated chitin and chitosan as novel biomaterials. Int J Biol Macromol, 2007, 40: 175–181CrossRefGoogle Scholar
  7. 7.
    Mariappan MR, Alas EA, Williams JG, Prager MD. Chitosan and chitosan sulfated have opposing effects on collagen-fibroblast interactions. Wound Repair Regen, 1999, 7: 400–406CrossRefGoogle Scholar
  8. 8.
    Li QL, Huang N, Chen JL, Chen C, Chen JY. Endothelial cell and platelet behavior on titanium modified with a mutilayer of polyelectrolytes. J Bioact Compat Pol, 2009, 24: 129–150CrossRefGoogle Scholar
  9. 9.
    Terbojevich M, Carraro C, Cosani A. Solution studies of chitosan 6-O-sulfate. Makromol Chem, 1989, 190: 2847–2855CrossRefGoogle Scholar
  10. 10.
    Tian Q, Wang XH, Wang W, Zhang CN, Wang P, Yuan Z. Self-assembly and liver targeting of sulfated chitosan nanoparticles functionalized with glycyrrhetinic acid. Nanomedicine, 2012, 8: 870–879Google Scholar
  11. 11.
    LeRoux MA, Guilak F, Setton LA. Compressive and shear properties of alginate gel: Effects of sodium ions and alginate concentration. J Biomed Mater Res, 1999, 47: 46–53CrossRefGoogle Scholar
  12. 12.
    Lin RZ, Miao J, Dong SX. Synthesis and Properties of pH-sensitive Sulfated Chitosan(SCS) Hydrogels. J Mater Sci Eng, 2008, 26: 950–953Google Scholar
  13. 13.
    Park KM, Shin YM, Joung YK, Shin H, Park KD. In situ forming hydrogels based on tyramine conjugated 4-Arm-PPO-PEO via enzymatic oxidative reaction. Biomacromolecules, 2010, 11: 706–712CrossRefGoogle Scholar
  14. 14.
    Lee F, Chung JE, Kurisawa M. An injectable hyaluronic acidtyramine hydrogel system for protein delivery. J Control Release, 2009, 134: 186–193CrossRefGoogle Scholar
  15. 15.
    Kurisawa M, Chung JE, Yang YY, Gao SJ, Uyama H. Injectable biodegradable hydrogels composed of hyaluronic acid-tyramine conjugates for drug delivery and tissue engineering. Chem Commun, 2005, 14: 4312–4314CrossRefGoogle Scholar
  16. 16.
    Sakai S, Kawakami K. Synthesis and characterization of both ionically and enzymatically cross-linkable alginate. Acta Biomater, 2007, 3: 495–501CrossRefGoogle Scholar
  17. 17.
    Wang LS, Boulaire J, Chan PP, Chung JE, Kurisawa M. The role of stiffness of gelatin-hydroxyphenylpropionic acid hydrogels formed by enzyme-mediated crosslinking on the differentiation of human mesenchymal stem cell. Biomaterials, 2010, 31: 8608–8616CrossRefGoogle Scholar
  18. 18.
    Hirakawa K, Hashizume K, Hayashi T. Viscoelastic property of human brain-for the analysis of impact injury. No To Shinkei, 1981, 33: 1057–1065Google Scholar
  19. 19.
    Miller K, Chinzei K, Orssengo G, Bednarz P. Mechanical properties of brain tissue in-vivo: Experiment and computer simulation. J Biomech, 2000, 33: 1369–1376CrossRefGoogle Scholar
  20. 20.
    Evans DW, Moran EC, Baptista PM, Soker S, Sparks JL. Scale-dependent mechanical properties of native and decellularized liver tissue. Biomech Model Mechanobiol, 2012, DOI 10.1007/s10237-012-0426-3Google Scholar
  21. 21.
    Chen F, Tian M, Zhang DM, Wang JY, Wang QG, Yu XX, Zhang XH, Wan CX. Preparation and characterization of oxidized alginate covalently cross-linked galactosylated chitosan scaffold for liver tissue engineering. Mat Sci Eng C-Bio S, 2011, 32: 310–320CrossRefGoogle Scholar
  22. 22.
    Drury JL, Mooney DJ. Hydrogels for tissue engineering: scaffold design variables and applications. Biomaterials, 2003, 24: 4337–4351CrossRefGoogle Scholar
  23. 23.
    O’Brien FJ, Harley BA, Yannas IV, Gibson LJ. The effect of pore size on cell adhesion in collagen-GAG scaffolds. Biomaterials, 2005, 26: 433–441CrossRefGoogle Scholar
  24. 24.
    Lee F, Chung JE, Kurisawa M. An injectable enzymatically crosslinked hyaluronic acid-tyramine hydrogel system with independent tuning of mechanical strength and gelation rate. Soft Matter, 2008, 4: 880–887CrossRefGoogle Scholar
  25. 25.
    Rosiak J, Olejniczak J, Pekala W. Fast reaction of irradiated polymers-I. Crosslinking and degradation of polyvinylpyrrolidone. Int J Radiat Appl Instrum C Radiat Phys Chem, 1990, 36: 747–755CrossRefGoogle Scholar
  26. 26.
    Jonas SK, Riley PA, Willson RL. Hydrogel peroxide cytotoxicity. Low-temperature enhancement by ascorbate or reduced lipoate. Biochem J, 1989, 264: 651–655Google Scholar
  27. 27.
    Novosel EC, Kleinhans C, Kluger PJ. Vascularization is the key challenge in tissue engineering. Adv Drug Deliv Rev, 2011, 63: 300–311CrossRefGoogle Scholar
  28. 28.
    Ott HC, Matthiesen TS, Goh SK, Black LD, Kren SM, Netoff TI, Taylor DA. Perfusion-decellularized matrix: Using nature’s platform to engineer a bioartificial heart. Nat Med, 2008, 14: 213–221CrossRefGoogle Scholar
  29. 29.
    Yu J, Gu Y, Du KT, Miharda S, Sievers RE, Lee RJ. The effect of injected RGD modified alginate on angiogenesis and left ventricular function in a chronic rat infarct model. Biomaterials, 2009, 30: 751–756CrossRefGoogle Scholar
  30. 30.
    Cai K, Kong T, Wang L, Liu P, Yang W, Chen C. Regulation of endothelial cells migration on poly (D, L-lactic acid) films immobilized with collagen gradients. Colloids Surf B Biointerfaces, 2010, 79: 291–297CrossRefGoogle Scholar
  31. 31.
    Li J, Pan J, Zhang L, Yu Y. Culture of hepatocytes on fructose-modified chitosan scaffolds. Biomaterials, 2003, 24: 2317–2322CrossRefGoogle Scholar
  32. 32.
    Fu D, Han B, Dong W, Yang Z, Lv Y, Liu W. Effects of carboxymethyl chitosan on the blood system of rats. Biochem Biophys Res Commun, 2011, 408: 110–114CrossRefGoogle Scholar

Copyright information

© Science China Press and Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  1. 1.Key Laboratory of Functional Polymer Materials, Ministry of Education; Institute of Polymer ChemistryNankai UniversityTianjinChina

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