title: Effect of N-Acetylcysteine on Adipose-Derived Stem Cell and Autologous Fat Graft Survival in a Mouse Model.
abstract: BACKGROUND
  Autologous fat grafting is a popular reconstructive technique, but is limited by inconsistent graft retention. The authors examined whether a widely available, clinically safe antioxidant, N-acetylcysteine, could improve adipose-derived stem cell survival and graft take when added to tumescent solution during fat harvest.
  
  
  METHODS
  Inguinal fat pads were harvested from C57BL/6 mice using tumescent solution with or without N-acetylcysteine. Flow cytometric, proliferation, and differentiation assays were performed on isolated primary adipose-derived stem cells and 3T3-L1 preadipocytes treated with or without hydrogen peroxide and/or N-acetylcysteine. N-Acetylcysteine-treated or control grafts were injected under recipient mouse scalps and assessed by serial micro-computed tomographic volumetric analysis. Explanted grafts underwent immunohistochemical analysis.
  
  
  RESULTS
  In culture, N-acetylcysteine protected adipose-derived stem cells from oxidative stress and improved cell survival following hydrogen peroxide treatment. Combined exposure to both N-acetylcysteine and hydrogen peroxide led to a 200-fold increase in adipose-derived stem cell proliferation, significantly higher than with either agent alone. N-Acetylcysteine decreased differentiation of adipose-derived stem cells into mature adipocytes, as evidenced by decreased transcription of adipocyte differentiation markers and reduced Oil Red-O staining. In vivo, N-acetylcysteine treatment resulted in improved graft retention at 3 months compared with control (46 versus 17 percent; p = 0.027). N-Acetylcysteine-treated grafts demonstrated less fibrosis and inflammation, and a 33 percent increase in adipocyte density compared with controls (p < 0.001) that was not associated with increased vascularity.
  
  
  CONCLUSIONS
  These findings provide proof of principle for the addition of N-acetylcysteine to tumescent harvest solution in the clinical setting to optimize fat graft yields.
authors.affiliation: Halifax, Nova Scotia, Canada From the Divisions of Plastic and Reconstructive Surgery and Otolaryngology, IWK Health Centre, the Departments of Microbiology and Immunology, Pharmacology, and Pediatrics, and the Faculty of Medicine, Dalhousie University.
authors.forename: Joshua
authors.initials: J
authors.lastname: Gillis
authors.forename: Simon
authors.initials: S
authors.lastname: Gebremeskel
authors.forename: Kyle D
authors.initials: KD
authors.lastname: Phipps
authors.forename: Lori A
authors.initials: LA
authors.lastname: MacNeil
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Sinal
authors.forename: Brent
authors.initials: B
authors.lastname: Johnston
authors.forename: Paul
authors.initials: P
authors.lastname: Hong
authors.forename: Michael
authors.initials: M
authors.lastname: Bezuhly
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D017667:Adipocytes / Q000187:drug effects* / Q000637:transplantation*; D000273:Adipose Tissue / Q000637:transplantation; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004195:Disease Models, Animal; D005434:Flow Cytometry; D006085:Graft Survival; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011897:Random Allocation; D012016:Reference Values; D033581:Stem Cell Transplantation; D020858:Tissue and Organ Harvesting; D014182:Transplantation, Autologous
chemical_list: D000111:Acetylcysteine