In vitro evaluation of matrix metalloproteinases as predictive testing for nickel, a model sensitizing agent.
The identification of potential damage due to chemical exposure in the workplace is a major health and regulatory concern. Traditional tests that measure both sensitization and elicitation responses require the use of animals. An alternative to this widespread use of experimental animals could have a crucial impact on risk assessment, especially for the preliminary screening of new molecules. We developed an in vitro model for the screening of potential toxic compounds. Human keratinocytes (HaCat) were used as target cells while matrix metalloproteinases (MMP) were selected as responders because they are key enzymes involved in extracellular matrix (ECM) degradation in physiological and pathological conditions. Chemical exposure was performed using nickel sulphate as a positive tester. Nickel contact induced upregulation of MMP-2 and IL-8 mRNA production. Molecular activation occurred even at very low nickel concentrations even though no phenotypic changes were observed. MMP-9 accumulation was found in the medium of treated cells with respect to controls. These observations led to the hypothesis that even minimal exposure can accumulate transcriptional activity resulting in long-term clinical signs after contact. Our simple in vitro model can be applied as a useful preliminary complement to the animal studies to screen the effects of new potential toxic compounds.
Lamberti
Monica
M
Occupational Medicine Section, Department of Experimental Medicine, Medical School, II University of Naples, 80128 Naples, Italy.
Perfetto
Brunella
B
Costabile
Teresa
T
Canozo
Nunzia
N
Baroni
Adone
A
Liotti
Francesco
F
Sannolo
Nicola
N
Giuliano
Mariateresa
M
D000826:Animal Testing Alternatives; D015415:Biomarkers / Q000032:analysis; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003877:Dermatitis, Contact / Q000209:etiology; D009783:Dermatitis, Occupational / Q000209:etiology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D016209:Interleukin-8 / Q000032:analysis / Q000096:biosynthesis; D015603:Keratinocytes / Q000187:drug effects* / Q000201:enzymology; D020782:Matrix Metalloproteinases / Q000032:analysis* / Q000096:biosynthesis; D008954:Models, Biological; D009532:Nickel / Q000633:toxicity*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018675:Toxicity Tests / Q000379:methods*; D015854:Up-Regulation
D015415:Biomarkers; D016209:Interleukin-8; D012333:RNA, Messenger; C029938:nickel sulfate; D009532:Nickel; D020782:Matrix Metalloproteinases