title: Development of a novel method for decellularizing a nerve graft using a hypertonic sodium chloride solution.
abstract: OBJECTIVE
  Acellular nerves are a reconstruction material and provide scaffolds for nerve regeneration. Numerous methods to develop acellular nerves have been described. However, these methods pose problems that can be attributed to incomplete acellular processing and destruction of the extracellular matrix (ECM); the former may lead to rejection response, while the latter may damage the scaffold. In order to overcome problems associated with the above-mentioned methods, we developed a novel method that employs a hypertonic sodium chloride solution to decellularize nerve graft material.
  
  
  METHODS
  Rat sciatic nerves were harvested, dipped in hypertonic sodium chloride solution (1 M), and shaken for 24 h. We then washed the nerves in phosphate-buffered saline for 7 days with shaking and evaluated the acellular nerves by hematoxylin-eosin (H-E) staining, immunostaining, and electron microscopy. We then transplanted the grafts to the sciatic nerve of another rat and evaluated the outcomes by H-E staining, immunostaining (anti-neurofilament antibody, anti-S-100 antibody), anterograde nerve tracing, and electron microscopy.
  
  
  RESULTS
  We found that our method successfully decellularized the grafts, but was mild enough to leave the ECM intact. Two months after transplantation, immunostaining and anterograde nerve tracing confirmed that Schwann cells infiltrated the grafts and induced neurofilament extension.
  
  
  CONCLUSIONS
  Our methodology preserves the ECM, is simple to develop, and does not involve substances that harm biogenic tissue. Acellular nerve tissue processed in this way could become a substitute material for bridging nerve gaps. Our method could also aid in the development of other acellular tissues.
authors.affiliation: 1 Department of Plastic and Reconstructive Surgery, Kobe University Graduate School of Medicine, Hyogo - Japan.
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Ishida
authors.forename: Shuhsuke
authors.initials: S
authors.lastname: Sakakibara
authors.forename: Hiroto
authors.initials: H
authors.lastname: Terashi
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Hashikawa
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamaoka
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D005109:Extracellular Matrix / Q000648:ultrastructure; D005260:Female; D007382:Intermediate Filaments / Q000378:metabolism; D023421:Models, Animal; D009416:Nerve Regeneration; D016067:Nerve Transfer; D017208:Rats, Wistar; D009418:S100 Proteins / Q000378:metabolism; D012462:Saline Solution, Hypertonic / Q000494:pharmacology*; D012584:Sciatic Nerve / Q000187:drug effects* / Q000378:metabolism / Q000637:transplantation* / Q000648:ultrastructure; D013997:Time Factors
chemical_list: D015415:Biomarkers; D009418:S100 Proteins; D012462:Saline Solution, Hypertonic