Frontiers in immunology · 2026
Evaluation of a multi-component mucoadhesive buccal patch: cytokine-associated inflammatory responses and tissue remodeling in experimental models.
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Kokkanti Rekha Rani, Viswanathan Sandhiya, Parida Nandita, Biswas Soumyajit, Kushwaha Gajraj Singh, Deheri Pratap Kumar, Panda Alok Kumar, Panda Abikshyeet, Bajoria Atul Anand, Patnaik Srinivas
Аннотация
INTRODUCTION: Dysregulated cytokine signaling is implicated in persistent inflammation and fibrosis in oral submucous fibrosis (OSMF) and has been reported in association with pathways observed in oral squamous cell carcinoma (OSCC). This study investigated cytokine associated inflammatory and fibrotic responses following treatment with a mucoadhesive buccal patch containing isotretinoin, bromelain, and limonene.
METHODS: The formulation was evaluated using lipopolysaccharide (LPS) induced acute inflammatory in vitro models and a carbon tetrachloride (CCl₄) induced in vivo oral fibrotic model. Gene expression analysis was performed using qPCR to assess IL-6, TNF-α, MMP-2, IL-2, and TGF-β expression. Protein expression of COX-2 and CK17 was also examined. Bromelain activity within the formulation was evaluated using collagen degradation assays under different concentrations and incubation periods. Histological analysis and functional assessment of mouth opening were conducted in vivo to evaluate fibrotic alterations.
RESULTS: LPS stimulation induced upregulation of IL-6, TNF-α, and MMP-2, while treatment with the formulation was associated with reduced expression of these inflammatory markers. Altered IL-2 and TGF-β expression suggested modulation of immune regulatory and fibrosis-associated pathways. The in vitro model represented an acute inflammatory epithelial response and did not fully recapitulate the fibrotic pathology of OSMF. Protein analysis demonstrated reduced COX-2 expression and alterations in CK17 levels, consistent with changes in inflammatory and epithelial responses. Bromelain retained enzymatic activity within the formulation and demonstrated concentration and time dependent collagen degradation. In vivo treatment showed reduced collagen-associated staining, improved tissue architecture, and changes in mouth opening, indicating attenuation of fibrotic progression.
DISCUSSION: These findings provide preliminary observations on cytokine-associated responses under inflammatory and fibrotic conditions relevant to OSMF. Further studies are required to define underlying mechanisms and evaluate translational applicability.
METHODS: The formulation was evaluated using lipopolysaccharide (LPS) induced acute inflammatory in vitro models and a carbon tetrachloride (CCl₄) induced in vivo oral fibrotic model. Gene expression analysis was performed using qPCR to assess IL-6, TNF-α, MMP-2, IL-2, and TGF-β expression. Protein expression of COX-2 and CK17 was also examined. Bromelain activity within the formulation was evaluated using collagen degradation assays under different concentrations and incubation periods. Histological analysis and functional assessment of mouth opening were conducted in vivo to evaluate fibrotic alterations.
RESULTS: LPS stimulation induced upregulation of IL-6, TNF-α, and MMP-2, while treatment with the formulation was associated with reduced expression of these inflammatory markers. Altered IL-2 and TGF-β expression suggested modulation of immune regulatory and fibrosis-associated pathways. The in vitro model represented an acute inflammatory epithelial response and did not fully recapitulate the fibrotic pathology of OSMF. Protein analysis demonstrated reduced COX-2 expression and alterations in CK17 levels, consistent with changes in inflammatory and epithelial responses. Bromelain retained enzymatic activity within the formulation and demonstrated concentration and time dependent collagen degradation. In vivo treatment showed reduced collagen-associated staining, improved tissue architecture, and changes in mouth opening, indicating attenuation of fibrotic progression.
DISCUSSION: These findings provide preliminary observations on cytokine-associated responses under inflammatory and fibrotic conditions relevant to OSMF. Further studies are required to define underlying mechanisms and evaluate translational applicability.