title: The impact of detergents on the tissue decellularization process: A ToF-SIMS study.
abstract: Biologic scaffolds are derived from mammalian tissues, which must be decellularized to remove cellular antigens that would otherwise incite an adverse immune response. Although widely used clinically, the optimum balance between cell removal and the disruption of matrix architecture and surface ligand landscape remains a considerable challenge. Here we describe the use of time of flight secondary ion mass spectroscopy (ToF-SIMS) to provide sensitive, molecular specific, localized analysis of detergent decellularized biologic scaffolds. We detected residual detergent fragments, specifically from Triton X-100, sodium deoxycholate and sodium dodecyl sulphate (SDS) in decellularized scaffolds; increased SDS concentrations from 0.1% to 1.0% increased both the intensity of SDS fragments and adverse cell outcomes. We also identified cellular remnants, by detecting phosphate and phosphocholine ions in PAA and CHAPS decellularized scaffolds. The present study demonstrates ToF-SIMS is not only a powerful tool for characterization of biologic scaffold surface molecular functionality, but also enables sensitive assessment of decellularization efficacy.
  
  
  
  We report here on the use of a highly sensitive analytical technique, time of flight secondary ion mass spectroscopy (ToF-SIMS) to characterize detergent decellularized scaffolds. ToF-SIMS detected cellular remnants and residual detergent fragments; increased intensity of the detergent fragments correlated with adverse cell matrix interactions. This study demonstrates the importance of maintaining a balance between cell removal and detergent disruption of matrix architecture and matrix surface ligand landscape. This study also demonstrates the power of ToF-SIMS for the characterization of decellularized scaffolds and capability for assessment of decellularization efficacy. Future use of biologic scaffolds in clinical tissue reconstruction will benefit from the fundamental results described in this work.
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: White
authors.affiliation: National ESCA and Surface Analysis Center for Biomedical Problems, University of Washington, Seattle, WA, United States; Department of Bioengineering, University of Washington, Seattle, WA, United States.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Taylor
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States; Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: Denver M
authors.initials: DM
authors.lastname: Faulk
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States; Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Keane
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States; Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: Lindsey T
authors.initials: LT
authors.lastname: Saldin
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: Janet E
authors.initials: JE
authors.lastname: Reing
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: Ilea T
authors.initials: IT
authors.lastname: Swinehart
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States.
authors.forename: Neill J
authors.initials: NJ
authors.lastname: Turner
authors.affiliation: National ESCA and Surface Analysis Center for Biomedical Problems, University of Washington, Seattle, WA, United States; Department of Bioengineering, University of Washington, Seattle, WA, United States.
authors.forename: Buddy D
authors.initials: BD
authors.lastname: Ratner
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States; Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, United States; Department of Surgery, University of Pittsburgh, Pittsburgh, PA, United States. Electronic address: badylaks@upmc.edu.
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Badylak
mesh_terms: D000818:Animals; D003902:Detergents / Q000737:chemistry*; D005109:Extracellular Matrix / Q000737:chemistry*; D013552:Swine; D001743:Urinary Bladder / Q000737:chemistry*
chemical_list: D003902:Detergents
keywords: Biologic scaffold; Decellularization; Detergents; Extracellular matrix; ToF-SIMS