title: Functionalization of titanium surface with chitosan via silanation: 3D CLSM imaging of cell biocompatibility behaviour.
abstract: BACKGROUND
  Biocompatibility ranks as one of the most important properties of dental materials. One of the criteria for biocompatibility is the absence of material toxicity to cells, according to the ISO 7405 and 10993 recommendations. Among numerous available methods for toxicity assessment; 3-dimensional Confocal Laser Scanning Microscopy (3D CLSM) imaging was chosen because it provides an accurate and sensitive index of living cell behavior in contact with chitosan coated tested implants.
  
  
  OBJECTIVE
  The purpose of this study was to investigate the in vitro biocompatibility of functionalized titanium with chitosan via a silanation using sensitive and innovative 3D CLSM imaging as an investigation method for cytotoxicity assessment.
  
  
  METHODS
  The biocompatibility of four samples (controls cells, TA6V, TA6V-TESBA and TA6V-TESBAChitosan) was compared in vitro after 24h of exposure. Confocal imaging was performed on cultured human gingival fibroblast (HGF1) like cells using Live/Dead® staining. Image series were obtained with a FV10i confocal biological inverted system and analyzed with FV10-ASW 3.1 Software (Olympus France).
  
  
  RESULTS
  Image analysis showed no cytotoxicity in the presence of the three tested substrates after 24 h of contact. A slight decrease of cell viability was found in contact with TA6V-TESBA with and without chitosan compared to negative control cells.
  
  
  CONCLUSIONS
  Our findings highlighted the use of 3D CLSM confocal imaging as a sensitive method to evaluate qualitatively and quantitatively the biocompatibility behavior of functionalized titanium with chitosan via a silanation. The biocompatibility of the new functionalized coating to HGF1 cells is as good as the reference in biomedical device implantation TA6V.
authors.affiliation: Université de Lyon. nina.attik@univ-lyon1.fr.
authors.forename: G N
authors.initials: GN
authors.lastname: Attik
authors.forename: M
authors.initials: M
authors.lastname: D'Almeida
authors.forename: B
authors.initials: B
authors.lastname: Toury
authors.forename: B
authors.initials: B
authors.lastname: Grosgogeat
mesh_terms: D000447:Aldehydes / Q000737:chemistry; D000497:Alloys; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D048271:Chitosan / Q000737:chemistry*; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D003722:Dental Alloys / Q000737:chemistry; D003764:Dental Materials / Q000737:chemistry*; D005347:Fibroblasts / Q000166:cytology; D005881:Gingiva / Q000166:cytology; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods; D008422:Materials Testing; D018613:Microscopy, Confocal / Q000379:methods; D012821:Silanes / Q000737:chemistry*; D013499:Surface Properties; D014025:Titanium / Q000737:chemistry*
chemical_list: D000447:Aldehydes; D000497:Alloys; D020099:Coated Materials, Biocompatible; D003722:Dental Alloys; D003764:Dental Materials; D012821:Silanes; C031462:titanium alloy (TiAl6V4); D048271:Chitosan; D014025:Titanium; C018475:butyraldehyde