The Impact of Adipose-derived Stem Cells (ASCs) on micro-RNAs of Keratinocytes in Oral Lichen Planus (OLP) patients using a 3D culture system: a Pilot Study

Background: Oral Lichen Planus (OLP) is a potentially malignant disorder with a neoplastic transformation risk of 0.07%-5.8%, lacking a definitive therapy. Adipose-derived stem cells (ASCs) exhibit immunomodulatory, pro-angiogenic, and oncosuppressive properties, potentially mediated by microRNAs (miRNAs), which may influence OLP progression. Aim: This pilot study investigates the oncosuppressive effects of allogeneic ASCs on OLP by analyzing miRNA expression and cellular behavior in keratinocytes using 2D/3D culture systems (organoids). Methods: Biopsies from 10 OLP patients and 10 healthy controls will be collected. One portion of the biopsy will be fixed for histopathological analysis, while a 1 mm fresh sample of the same biopsy, will be used for keratinocyte isolation and 2D/3D culture. Keratinocytes will be co-cultured with allogeneic ASCs from healthy donors or with ASC secretome to assess changes in proliferation, migration, vitality, and miRNA expression (miR-375, miR-372, miR- 10b) via qRT-PCR and Western Blot. A 3D microfluidic model mimicking oral mucosa will validate 2D findings, evaluating ASC effects on fibrosis, extracellular matrix, and neoplastic transformation markers. Results: ASCs’ secretome significantly reduce oncogenic miRNA expression and inhibit proliferation and migration in OLP keratinocytes, potentially via paracrine mechanisms, with the organoid model confirming these effects in a near-in vivo setting. Conclusion: This study aims to elucidate the therapeutic potential of ASCs in OLP and paving the way for cell-free therapeutic approaches, showing that cell therapy turns off chronic inflammation of the OLP at the molecular level, lowering the risk of carcinomatous evolution.