title: Quantitative stain-free and continuous multimodal monitoring of wound healing in vitro with digital holographic microscopy.
abstract: Impaired epithelial wound healing has significant pathophysiological implications in several conditions including gastrointestinal ulcers, anastomotic leakage and venous or diabetic skin ulcers. Promising drug candidates for accelerating wound closure are commonly evaluated in in vitro wound assays. However, staining procedures and discontinuous monitoring are major drawbacks hampering accurate assessment of wound assays. We therefore investigated digital holographic microscopy (DHM) to appropriately monitor wound healing in vitro and secondly, to provide multimodal quantitative information on morphological and functional cell alterations as well as on motility changes upon cytokine stimulation. Wound closure as reflected by proliferation and migration of Caco-2 cells in wound healing assays was studied and assessed in time-lapse series for 40 h in the presence of stimulating epidermal growth factor (EGF) and inhibiting mitomycin c. Therefore, digital holograms were recorded continuously every thirty minutes. Morphological changes including cell thickness, dry mass and tissue density were analyzed by data from quantitative digital holographic phase microscopy. Stimulation of Caco-2 cells with EGF or mitomycin c resulted in significant morphological changes during wound healing compared to control cells. In conclusion, DHM allows accurate, stain-free and continuous multimodal quantitative monitoring of wound healing in vitro and could be a promising new technique for assessment of wound healing.
authors.affiliation: Department of Medicine B, University Hospital Münster, Münster, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Bettenworth
authors.affiliation: Department of Medicine B, University Hospital Münster, Münster, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Lenz
authors.affiliation: Department of Medicine B, University Hospital Münster, Münster, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Krausewitz
authors.affiliation: Department of Medicine B, University Hospital Münster, Münster, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Brückner
authors.affiliation: Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany; Biomedical Technology Center, University of Münster, Münster, Germany.
authors.forename: Steffi
authors.initials: S
authors.lastname: Ketelhut
authors.affiliation: Department of Medicine B, University Hospital Münster, Münster, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Domagk
authors.affiliation: Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany; Biomedical Technology Center, University of Münster, Münster, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Kemper
mesh_terms: D000465:Algorithms; D018938:Caco-2 Cells; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006696:Holography / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D016685:Mitomycin / Q000494:pharmacology; D008954:Models, Biological; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D015203:Reproducibility of Results; D014945:Wound Healing / Q000187:drug effects / Q000502:physiology*
chemical_list: D019384:Nucleic Acid Synthesis Inhibitors; D016685:Mitomycin; D004815:Epidermal Growth Factor