Co-Transfection With Anti-Fibrotic microRNAs for Treating Oral Submucous Fibrosis.

Area of Science:

• Biomedical research
• Molecular biology
• Oral pathology

Background:

• Oral submucous fibrosis (OSF) involves excessive collagen deposition, linked to betel nut use.
• Arecoline in betel nut activates the TGF-β/Smads pathway, promoting fibrosis.
• MicroRNA (miR) dysregulation contributes to OSF, making miR modulation a potential therapy.

Purpose of the Study:

• To investigate the therapeutic potential of co-transfecting multiple anti-fibrotic miRs for OSF.
• To compare the efficacy of combined miR transfection versus single miR transfection in an OSF model.

Main Methods:

• Human oral fibroblasts were stimulated with arecoline.
• 16 miRs were individually transfected, with miR-29a-3p, miR-196a-3p, and miR-509-5p selected for co-transfection based on their ability to downregulate TGFB1 and ACTA2.
• Co-transfection efficacy was assessed via mRNA and protein expression analysis, and functional assays like wound closure and collagen gel contraction.

Main Results:

• Co-transfection significantly downregulated key fibrotic genes (COL1A1, COL3A1, COL5A1, MMP1, MMP7, TIMP1, TIMP2).
• Combined miR transfection showed superior downregulation of COL1A1 and MMP1 compared to single miRs.
• The treatment suppressed the TGF-β/Smads pathway, reduced extracellular matrix production, and inhibited fibroblast migration and contraction.

Conclusions:

• Co-transfection of selected anti-fibrotic miRs offers a promising therapeutic strategy for oral submucous fibrosis.
• This multi-miR approach demonstrates enhanced efficacy in combating fibrosis compared to single miR treatments.