title: [Effect of carboxymethylated chitosan on proliferation and synthesis of neurotrophic factors in Schwann cells in vitro].
abstract: OBJECTIVE
  To investigate the effect of carboxymethylated chitosan (CMCS) on the proliferation, cell cycle, and secretion of neurotrophic factors in cultured Schwann cells (SCs).
  
  
  METHODS
  SCs were obtained from sciatic nerves of 20 Sprague Dawley rats (3-5 days old; male or female; weighing, 25-30 g) and cultured in vitro, SCs were identified and purified by immunofluorescence against S-100. The cell counting kit 8 (CCK-8) assay was used to determine the proliferation of SCs. The SCs were divided into 4 groups: 50 microg/mL CMCS (group B), 100 microg/mL CMCS (group C), 200 microg/mL CMCS (group D), and the same amount of PBS (group A) were added. The flow cytometry was used to analyze the cell cycle of SCs; the real-time quantitative PCR and Western blot analysis were used to detect the levels of never growth factor (NGF) and ciliary neurotrophic factor (CNTF) in cultured SCs induced by CMCS.
  
  
  RESULTS
  The purity of cultured SCs was more than 90% by immunofluorescence against S-100; the CCK-8 results indicated that CMCS in concentrations of 10-1 000 microg/mL could promote the proliferation of SCs, especially in concentrations of 200 and 500 microg/mL (P < 0.01), but no significant difference was found between 200 and 500 microg/mL (P > 0.05). CMCS at a concentration of 200 microg/mL for 24 hours induced the highest proliferation, showing significant difference when compared with that at 0 hour (P < 0.01). The percentage of cells in phase S and the proliferation index were significantly higher in groups B, C, and D than in group A (P < 0.05), in groups C and D than in group B (P < 0.05); and there was no significant difference between group C and group D (P > 0.05). Real-time quantitative PCR and Western blot results showed that the levels of NGF and CNTF in groups B, C, and D were significantly higher than those in group A (P < 0.05), especially in group D.
  
  
  CONCLUSIONS
  CMCS can stimulate the proliferation, and induce the synthesis of neurotrophic factors in cultured SCs.
authors.affiliation: Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan Hubei, 430060, P R China. hebin@whu.edu.cn
authors.forename: Bin
authors.initials: B
authors.lastname: He
authors.forename: Haiying
authors.initials: H
authors.lastname: Tao
authors.forename: Ailin
authors.initials: A
authors.lastname: Wei
authors.forename: Shiqing
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D048271:Chitosan / Q000031:analogs & derivatives* / Q000494:pharmacology; D005260:Female; D008297:Male; D009414:Nerve Growth Factors / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D012584:Sciatic Nerve / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods
chemical_list: D009414:Nerve Growth Factors; D012333:RNA, Messenger; D048271:Chitosan