Basement Membrane-Derived Peptide Nanocoatings Promote Junctional Epithelium-Like Keratinocyte Function
Objectives: Millions of percutaneous devices (such as dental implants and restorations) fail from infection yearly. Ballooning antibiotic usage fuels antimicrobial resistance and is ineffective. Preventative strategies overlook soft tissue adhesion to prevent infection at the tissue-implant interface. Teeth, long-lasting percutaneous organs, feature soft tissue attachment through adhesive structures, hemidesmosomes (HD), in the junctional epithelium basement membrane adjacent to teeth. Inspired by this, we hypothesized cell adhesion peptide (CAPs) nanocoatings could promote oral keratinocyte (OK) HD formation and recapitulate junctional epithelium functions.
Methods: Four CAPs curated from basement membrane biomolecules were used to synthesize nanocoatings and controls using click chemistry and were physicochemically characterized, including after simulated oral durability challenges. Immunofluorescent imaging monitored OK HD formation dynamics, downstream kinase activity, and expression of junctional epithelium markers. Fibroblast mono-culture and a fibroblast/OK co-culture were used to understand the cell-specific attachment a subset of nanocoatings exhibited. Integrin blocking antibodies were used to probe cell-CAP interactions. Finally, a cytokine array assessed the effect of nanocoatings on cytokine expression. Statistical significance was determined by appropriate tests, p<0.05.
Results: Two CAP nanocoatings markedly increased HD formation, associated kinase activity, and upregulated expression of junctional epithelium functional biomolecules. Integrin blocking demonstrated multiple integrin heterodimers can synergistically contribute to initial cell adherence, early proliferation, and HD signaling. This control over integrin engagement – namely integrin α6β4 – was further demonstrated by cell-specific proliferation of OKs compared to fibroblasts and their differential integrin expression profiles. These nanocoatings simultaneously elicited a pro-reparative OK cytokine profile compared to controls.
Conclusions: Basement membrane-derived CAP nanocoatings were developed to recapitulate hallmark features of the oral junctional epithelium toward reducing infection. These nanocoatings provide a framework to form regenerative microenvironmental niches, study cell-matrix interplay, and improve biomaterial-based therapies.