Synthesis, characterisation and preliminary investigation of the haemocompatibility of polyethyleneimine-grafted carboxymethyl chitosan for gene delivery

Mater Sci Eng C Mater Biol Appl. 2016 May:62:173-82. doi: 10.1016/j.msec.2016.01.050. Epub 2016 Jan 22.

Abstract

The development of safe and efficient gene carriers is the key to the clinical success of gene therapy. In the present study, carboxymethyl chitosan (CMCS) was prepared by chitosan (CS) alkalisation and carboxymethylation reactions. Then polyethyleneimine (PEI) was grafted to the backbone of CMCS by an amidation reaction. The CMCS-PEI copolymer showed strong complexation capability with DNA to form nanoparticles, and achieved lower cytotoxicity and higher transfection efficiency compared with PEI (25 kDa) towards 293T and 3T3 cells. Moreover, the haemocompatibility of the CMCS-PEI copolymer was investigated through the aggregation, morphology and lysis of human red blood cells (RBCs), along with the impact on the clotting function with activated partial thromboplastin time (APTT), prothrombin time (PT) and thromboelastographic (TEG) assays. The results demonstrated that the CMCS-PEI copolymer with a concentration lower than 0.05 mg/mL had little impact on the aggregation, morphology or lysis of RBCs, or on blood coagulation. Therefore, the copolymer may be a strong alternative candidate as an effective and safe non-viral vector.

Keywords: Biocompatibility; Chitosan; Gene carrier; Haemocompatibility; Polyethyleneimine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biocompatible Materials / chemistry*
  • Biocompatible Materials / toxicity
  • Cell Line
  • Cell Survival / drug effects
  • Chitosan / analogs & derivatives*
  • Chitosan / chemistry
  • Chitosan / toxicity
  • Electrophoresis, Agar Gel
  • Erythrocytes / cytology
  • Erythrocytes / drug effects
  • HEK293 Cells
  • Hemolysis / drug effects
  • Humans
  • Mice
  • Microscopy, Electron, Transmission
  • Partial Thromboplastin Time
  • Particle Size
  • Plasmids / metabolism
  • Polyethyleneimine / analogs & derivatives*
  • Polyethyleneimine / chemistry
  • Polyethyleneimine / toxicity
  • Prothrombin Time
  • Thrombelastography
  • Transfection*

Substances

  • Biocompatible Materials
  • chitosan-graft-polyethylenimine
  • Polyethyleneimine
  • Chitosan