title: In vitro cell culture system optimization of keratinocytes from oral lichen planus (OLP) patients.
abstract: OBJECTIVE
  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.
  
  
  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 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.
authors.affiliation: Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.|Department of Endodontics and Oral Medicine, Nantong Stomatological Hospital, Nantong, China.
authors.forename: T
authors.initials: T
authors.lastname: Cao
authors.affiliation: Department of Clinical Immunology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: L
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: G
authors.initials: G
authors.lastname: Du
authors.affiliation: Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: H
authors.initials: H
authors.lastname: Yao
authors.affiliation: Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Clinical Immunology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Luo
authors.affiliation: Department of Clinical Immunology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: F
authors.initials: F
authors.lastname: Chen
authors.affiliation: Department of Periodontology and Oral Medicine, Qingdao Stomatological Hospital, Qingdao, China.
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: G
authors.initials: G
authors.lastname: Tang
mesh_terms: D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D000935:Antifungal Agents / Q000627:therapeutic use*; D016022:Case-Control Studies; D049109:Cell Proliferation; D016207:Cytokines / Q000378:metabolism*; D005260:Female; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism / Q000648:ultrastructure; D007633:Keratins / Q000378:metabolism; D017676:Lichen Planus, Oral / Q000473:pathology*; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D061251:Primary Cell Culture / Q000379:methods*; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism; D055815:Young Adult
chemical_list: D000935:Antifungal Agents; D016207:Cytokines; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D051996:Transcription Factor RelA; D014746:Vimentin; D007633:Keratins
keywords: NF-κB; TLR4; culture system; keratinocytes; oral lichen planus