Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (47): 7655-7659.doi: 10.3969/j.issn.2095-4344.2015.47.020

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Anticoagulant chitosan nanoparticles: synthesis, characterization and biological safety

Wang Zi-jing1, Liu Chao-yang1, Qian Zhi-yong2, Gao De-wei1, Guo Xi-min2   

  1. 1Department of Integrated Surgery, South Building, Chinese PLA General Hospital, Beijing 100853, China; 2Academy of Military Medical Sciences of Chinese PLA, Beijing 100040, China
  • Received:2015-08-25 Online:2015-11-19 Published:2015-11-19
  • Contact: Gao De-wei, Chief physician, Department of Integrated Surgery, South Building, Chinese PLA General Hospital, Beijing 100853, China Guo Xi-min, Associate researcher, Academy of Military Medical Sciences of Chinese PLA, Beijing 100040, China
  • About author:Wang Zi-jing, Studying for master’s degree, Department of Integrated Surgery, South Building, Chinese PLA General Hospital, Beijing 100853, China

Abstract:

BACKGROUND: Studies have shown that chitosan and other natural polysaccharides have heparin-like anticoagulant function after sulfonated modification. Sulfonated chitosan has good anticoagulant property because the sulfonate group formed by sulfonated chitosan is similar with the active group of heparin.

OBJECTIVE: To prepare the anticoagulant chitosan nanoparticles and to detect its morphology, physical and chemical properties and biological security.
METHODS: Chitosan nanoparticles were synthesized by emulsion-chemical cross link. Sulfonated chitosan nanoparticles were synthesized by sulfonation reaction. Its morphology was described by transmission electron microscope. The peak-value change of its specific groups was observed by infrared spectroscopy. (1) Coagulation experiment: Heparin, chitosan nanoparticles and 10, 30 and 50 mg of sulfonated chitosan nanoparticles were added into the blood of Spraque-Dawley rats. The coagulation indicators were detected. (2) Hemolysis experiment: deionized water, physiological saline and 10, 30, 50 g/L sulfonated chitosan nanoparticles extracts were added into 2% red blood cell suspension of rabbits. The hemolysis rate was detected. (3) Cytotoxicity experiments: DMEM medium containing fetal bovine serum and 10, 30, 50 g/L sulfonated chitosan nanoparticle extracts were used to culture human umbilical vein endothelial cells. Cell relative growth rate and toxicity grading were detected after 72 hours.
RESULTS AND CONCLUSION: Scanning electron microscopy showed that sulfonated chitosan nanoparticles had good morphology, with a diameter of 50 nm. Infrared spectroscopy showed that the sulfonated replacement occurred. In vitro coagulation experiments showed that sulfonated chitosan nanoparticles had significant anticoagulant effects in a dose-dependent manner. Sulfonated chitosan nanoparticles meet the national safety standard for hemolysis rate of less than 5%, non-induced hemolysis property. Cytotoxicity assays showed that sulfonated chitosan nanoparticles extracts had no significant cytotoxicity, and its biological safety was in line with the national standards. 
中国组织工程研究杂志出版内容重点:生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程

Key words: Chitosan, Nanostructures, Microspheres, Tissue Engineering