In Vitro Cell Culture System Optimization of Keratinocytes from Oral Lichen Planus (OLP) patients

Objectives The aim of this study was to optimize the culture system of keratinocytes obtained from oral lichen planus (OLP) patients and verify whether this model could simulate the local inflammatory environment of OLP. Materials and methods Keratinocytes were isolated from 48 OLP patients 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 TLR4 and NF-κB p65 expression, and the levels of IL-1β, 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 TLR4/NF-κB p65 and inflammatory cytokines, including IL-1β, 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. This article is protected by copyright. All rights reserved.