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


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.


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.



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.


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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

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