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In vitro cell culture system optimization of keratinocytes from oral lichen planus ( OLP ) patients
Author(s) -
Cao T,
Zhang H,
Zhou L,
Wang Y,
Du G,
Yao H,
Wang Y,
Luo Q,
Chen F,
Wang W,
Tang G
Publication year - 2017
Publication title -
oral diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.953
H-Index - 87
eISSN - 1601-0825
pISSN - 1354-523X
DOI - 10.1111/odi.12599
Subject(s) - oral lichen planus , vimentin , in vitro , keratinocyte , immunohistochemistry , cell culture , staining , pathology , chemistry , biology , microbiology and biotechnology , medicine , biochemistry , genetics
Objectives The aim of this study was to optimize the culture system of keratinocytes obtained from patients with oral lichen planus ( OLP ) and verify whether this model could simulate the local inflammatory environment of OLP . Materials and methods Keratinocytes were isolated from 48 patients with OLP and cultured in vitro . The ultrastructure of OLP keratinocytes was observed via electron microscopy. The expression of pancytokeratin and vimentin was determined by immunohistochemistry, and the proliferation of OLP keratinocytes was measured by CCK ‐8 assay. Immunofluorescence staining was used to detect TLR 4 and NF ‐ κ B p65 expression, and the levels of IL ‐1 β , IL ‐6, and TNF ‐ α in the supernatant were measured by ELISA . Results When seeded in plates precoated with recombinant human type‐1 collagen, keratinocytes isolated from patients who received systemic antifungal treatment and were younger than 40 years were more successful to be cultured in vitro . Characteristic pancytokeratin was expressed in almost all OLP keratinocytes. Compared with normal oral keratinocytes, OLP keratinocytes demonstrated higher levels of TLR 4/ NF ‐ κ B p65 and inflammatory cytokines, including IL ‐1 β , IL ‐6, and TNF ‐ α . Conclusions We successfully optimized the culture system of OLP keratinocytes,which mimicked the local inflammatory environment of OLP and may be used as a cell model of OLP .