Advertisement

Preparation, fabrication and biocompatibility of novel injectable temperature-sensitive chitosan/glycerophosphate/collagen hydrogels

  • Kedong Song
  • Mo Qiao
  • Tianqing Liu
  • Bo Jiang
  • Hugo M. Macedo
  • Xuehu Ma
  • Zhanfeng Cui
Article

Abstract

This paper introduces a novel type of injectable temperature-sensitive chitosan/glycerophosphate/collagen (C/GP/Co) hydrogel that possesses great biocompatibility for the culture of adipose tissue-derived stem cells. The C/GP/Co hydrogel is prepared by mixing 2.2% (v/v) chitosan with 50% (w/w) β-glycerophosphate at different proportions and afterwards adding 2 mg/ml of collagen. The gelation time of the prepared solution at 37°C was found to be of around 12 min. The inner structure of the hydrogel presented a porous spongy structure, as observed by scanning electron microscopy. Moreover, the osmolality of the medium in contact with the hydrogel was in the range of 310–330 mmol kg−1. These analyses have shown that the C/GP/Co hydrogels are structurally feasible for cell culture, while their biocompatibility was further examined. Human adipose tissue-derived stem cells (ADSCs) were seeded into the developed C/GP and C/GP/Co hydrogels (The ratios of C/GP and C/GP/Co were 5:1 and 5:1:6, respectively), and the cellular growth was periodically observed under an inverted microscope. The proliferation of ADSCs was detected using cck-8 kits, while cell apoptosis was determined by a Live/Dead Viability/Cytotoxicity kit. After 7 days of culture, cells within the C/GP/Co hydrogels displayed a typical adherent cell morphology and good proliferation with very high cellular viability. It was thus demonstrated that the novel C/GP/Co hydrogel herein described possess excellent cellular compatibility, representing a new alternative as a scaffold for tissue engineering, with the added advantage of being a gel at the body’s temperature that turns liquid at room temperature.

Keywords

Chitosan Gelation Time Cellular Viability Chitosan Molecule Tertiary Butyl Alcohol 
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.

Notes

Acknowledgments

This work was supported by the National Science Foundation of China (30670525, 30700181) and the new teacher foundation of Ministry of Education (20070141055). Mr. Hugo M. Macedo is also grateful to the Portuguese Fundação para a Ciência e Tecnologia for his PhD grant number SFRH/BD/28138/2006.

References

  1. 1.
    Tanzi MC, Farè S. Adipose tissue engineering: state of the art, recent advances and innovative approaches. Expert Rev Med Devices. 2009;6(5):533–51.CrossRefPubMedGoogle Scholar
  2. 2.
    Song K, Liu T, Cui Z, Li X, Ma X. Three-dimensional fabrication of engineered bone with human bio-derived bone scaffolds in a rotating wall vessel bioreactor. J Biomed Mater Res A. 2008;86(2):323–32.PubMedGoogle Scholar
  3. 3.
    Song K, Yang Z, Liu T, Zhi W, Li X, Deng L, Cui Z, Ma X. Fabrication and detection of tissue-engineered bones with bio-derived scaffolds in a rotating bioreactor. Biotechnol Appl Biochem. 2006;45(Pt 2):65–74.PubMedGoogle Scholar
  4. 4.
    Hutmacher DW, Schantz JT, Lam CX, Tan KC, Lim TC. State of the art and future directions of scaffold-based bone engineering from a biomaterials perspective. J Tissue Eng Regen Med. 2007;1(4):245–60.CrossRefPubMedGoogle Scholar
  5. 5.
    Bonzani IC, Adhikari R, Houshyar S, Mayadunne R, Gunatillake P, Stevens MM. Synthesis of two-component injectable polyurethanes for bone tissue engineering. Biomaterials. 2007;28(3):423–33.CrossRefPubMedGoogle Scholar
  6. 6.
    Choi GH, Youn YH, Kim DY, et al. Alginate gel decreases chondroitin sulfate proteoglycan immunoreactivity following spinal cord injury: preliminary study. Tissue Eng Reg Med. 2006;3(4):472–7.Google Scholar
  7. 7.
    Park H, Temenoff JS, Tabata Y, Caplan AI, Mikos AG. Injectable biodegradable hydrogel composites for rabbit marrow mesenchymal stem cell and growth factor delivery for cartilage tissue engineering. Biomaterials. 2007;28(21):3217–27.CrossRefPubMedGoogle Scholar
  8. 8.
    Chen L, Tian Z, Du Y. Synthesis and pH sensitivity of carboxymethyl chitosan-based polyampholyte hydrogels for protein carrier matrices. Biomaterials. 2004;25(17):3725–32.CrossRefPubMedGoogle Scholar
  9. 9.
    Hoemann CD, Sun J, Légaré A, McKee MD, Buschmann MD. Tissue engineering of cartilage using an injectable and adhesive chitosan-based cell-delivery vehicle. Osteoarthritis Cartilage. 2005;13(4):318–29.CrossRefPubMedGoogle Scholar
  10. 10.
    Crompton KE, Goud JD, Bellamkonda RV, et al. The polylysine-functionalised thermoresponsive chitosan hydrogel for neural tissue engineering. Biomaterials. 2007;28(3):441–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Cho JH, Kim SH, Park KD, et al. Chondrogenic differentiation of human mesenchymal stem cells using a thermosensitive poly (N-isopropylacrylamide) and water-soluble chitosan copolymer. Biomaterials. 2004;25(26):5743–51.CrossRefPubMedGoogle Scholar
  12. 12.
    Desai SD, Blanchard J. In vitro evaluation of Pluronic F127-based controlled-release ocular delivery systems for pilocarpine. J Pharm Sci. 1998;87(2):226–30.CrossRefPubMedGoogle Scholar
  13. 13.
    Qi-sheng Gu. Clinical application of collagen. Chin J Reparative Reconstruct Surg. 2006;20(10):1052–8.Google Scholar
  14. 14.
    Guo-ying L, Zhong-kai Z, Su L, Bi S. Differences in properties of collagen, gelatin and collagen hydrolysate. J Sichuan Univ Eng Sci Ed. 2005;37(4):54–8.Google Scholar
  15. 15.
    Geng X, Kwon OH, Jang J. Electrospinning of chitosan dissolved in concentrated acetic acid solution. Biomaterials. 2005;26(27):5427–32.CrossRefPubMedGoogle Scholar
  16. 16.
    Ma L, Shi Y, Chen Y, Zhao H, Gao C, Han C. In vitro and in vivo biological performance of collagen-chitosan/silicone membrane bilayer dermal equivalent. J Mater Sci Mater Med. 2007;18(11):2185–91.CrossRefPubMedGoogle Scholar
  17. 17.
    Wang L, Stegemann JP. Thermogelling chitosan and collagen composite hydrogels initiated with beta-glycerophosphate for bone tissue engineering. Biomaterials. 2010;31(14):3976–85.CrossRefPubMedGoogle Scholar
  18. 18.
    Rodriguez AM, Elabd C, Delteil F, Astier J, Vernochet C, Saint-Marc P, Guesnet J, Guezennec A, Amri EZ, Dani C, Ailhaud G. Adipocyte differentiation of multipotent cells established from human adipose tissue. Biochem Biophys Res Commun. 2004;315(2):255–63.CrossRefPubMedGoogle Scholar
  19. 19.
    Chenite A, Buschmann M, Wang D, Chaput C, Kandani N. Rheological characterisation of thermogelling chitosan/glycerol-phosphate solutions. Biomaterials. 2001;46(1):39–47.Google Scholar
  20. 20.
    Chenite A, Chaput C, Wang D, Combes C, Buschmann MD, Hoemann CD, Leroux JC, Atkinson BL, Binette F, Selmani A. Novel injectable neutral solutions of chitosan form biodegradable gels in situ. Biomaterials. 2000;21(21):2155–61.CrossRefPubMedGoogle Scholar
  21. 21.
    Chen RN, Wang GM, Chen CH, Ho HO, Sheu MT. Development of N,O-(carboxymethyl) chitosan/collagen matrixes as a wound dressing. Biomacromolecules. 2006;7(4):1058–64.CrossRefPubMedGoogle Scholar
  22. 22.
    Ma J, Wang H, He B, Chen J. A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetal dermal fibroblasts. Biomaterials. 2001;22(4):331–6.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Kedong Song
    • 1
  • Mo Qiao
    • 1
  • Tianqing Liu
    • 1
  • Bo Jiang
    • 1
  • Hugo M. Macedo
    • 2
  • Xuehu Ma
    • 1
  • Zhanfeng Cui
    • 3
  1. 1.Dalian R&D Center for Stem Cell and Tissue Engineering, State Key Laboratory of Fine ChemicalsDalian University of TechnologyDalianChina
  2. 2.Biological Systems Engineering Laboratory, Department of Chemical EngineeringImperial College LondonLondonUK
  3. 3.Oxford Centre for Tissue Engineering and BioprocessingUniversity of OxfordOxfordUK

Personalised recommendations