A time-lapse approach to examine chromium and nickel effects on wound healing in vitro.
Chromium and nickel cause allergic contact dermatitis, a common biological skin response to sensitizing agents. This study used a conventional in vitro wounding model to study the impact of sensitizing agents on the innate immune response of human keratinocytes. Experiments were designed to evaluate the involvement of specific Toll-like receptors and metalloproteinases as effectors molecules downstream, at a molecular level. Further, keratinocytes were co-cultured with monocytes (THP-1 cells) to reproduce an inductive stimulus on monocytes made by metals. Human keratinocytes (HaCat) were grown on plates covered with collagen type I, chemically treated, and then mechanically injured with a sterile pipette tip. Restoration of the monolayer integrity was monitored by time-lapse video microscopy. Effector gene expression was evaluated by real-time PCR. The presence of chromium significantly dropped the rate of wound closure, while nickel-induced hyper-proliferation ended in an acceleration of the healing process, an event that does not occur in vivo. This latter outcome led to considering nickel as an unsuitable example for use in the experimental model. Focusing solely on the chromium aspect of this study, RNA profiles of selected molecular markers were generated to ascertain if the detrimental stimulus from chromium was eliminated or persisted both in keratinocytes alone and/or during co-cultures of keratinocytes and monocytes. Monocytes accelerated the process of wound repair. This in vitro experimental model highlighted the involvement of innate immunity in response to chromium and might be useful for test molecules of therapeutic interest for the treatment of skin lesions. However, the experience with nickel reveals that there are limitations to the utility of this wound model system after all.
Microbiology and Clinical Microbiology Section, Naples, Italy.
Brunella
B
Perfetto
Antonietta
A
Stellavato
Alessandro
A
Melito
Vincenza
V
De Gregorio
Marcella
M
Cammarota
Mariateresa
M
Giuliano
D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000276:immunology; D002857:Chromium / Q000009:adverse effects*; D017449:Dermatitis, Allergic Contact / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects; D018836:Inflammation Mediators / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D009000:Monocytes / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D009532:Nickel / Q000009:adverse effects*; D059008:Time-Lapse Imaging / Q000379:methods*; D014945:Wound Healing / Q000187:drug effects* / Q000276:immunology
D018836:Inflammation Mediators; D002857:Chromium; D009532:Nickel