title: Comparison of therapeutic antibiotic treatments on tissue-engineered human skin substitutes.
abstract: For regenerative medicine to gain clinical acceptance, the effects of commonly used treatment regimens on bioengineered organs must be considered. The antibiotics mafenide acetate (mafenide) and neomycin plus polymyxin (neo/poly) are routinely used to irrigate postoperative skin grafts on contaminated wounds. The effects of these clinically used antibiotics were investigated using tissue-engineered human skin substitutes generated with primary human keratinocytes or the near-diploid human keratinocyte cell line, Near-diploid Immortal Keratinocytes. Following topical or dermal treatment, the skin substitutes were assayed for viability, tissue morphology, glycogen content, and the expression of active caspase 3. Mafenide, but not neo/poly, induced morphological and biochemical changes in tissue-engineered skin substitutes. Keratinocytes in all histological layers of mafenide-treated skin substitutes exhibited ballooning degeneration and glycogen depletion. Mafenide-treatment also triggered separation of basal keratinocytes from the underlying dermis. None of the antibiotic treatments induced apoptosis, as measured by active caspase 3 immunostaining. The results demonstrate that mafenide, but not neo/poly, is detrimental to the viability and structural integrity of tissue-engineered human skin substitutes. These findings highlight the need to identify treatment regimens that are compatible with and hence enable the therapeutic efficacy of first-generation bioengineered organs such as skin.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
authors.forename: Angela L
authors.initials: AL
authors.lastname: Gibson
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Schurr
authors.forename: Sandy J
authors.initials: SJ
authors.lastname: Schlosser
authors.forename: Allen R
authors.initials: AR
authors.lastname: Comer
authors.forename: B Lynn
authors.initials: BL
authors.lastname: Allen-Hoffmann
mesh_terms: D000900:Anti-Bacterial Agents / Q000009:adverse effects* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects; D001672:Biocompatible Materials; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D006003:Glycogen / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008272:Mafenide / Q000009:adverse effects / Q000627:therapeutic use; D008422:Materials Testing; D009355:Neomycin / Q000627:therapeutic use; D011113:Polymyxins / Q000627:therapeutic use; D017768:Skin, Artificial; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D000900:Anti-Bacterial Agents; D001672:Biocompatible Materials; D011113:Polymyxins; D008272:Mafenide; D006003:Glycogen; C505407:CASP3 protein, human; D053148:Caspase 3; D009355:Neomycin