Speaker
Description
"Introduction: Chronic ulcerative oral mucosal inflammatory diseases, including oral lichen planus and recurrent aphthous stomatitis, are painful and highly prevalent, yet lack effective clinical management. These conditions are usually treated with either topically applied or systemically-delivered corticosteroids depending on severity with varying degrees of success. In recent years systemic biologic therapies, including monoclonal antibodies that block the activity of proinflammatory cytokines, have increasingly been used to treat similar immune-mediated inflammatory conditions, such as rheumatoid arthritis and psoriasis. The ability to deliver these therapies locally to the oral epithelium could radically alter treatment options for oromucosal inflammatory diseases, where cytokines, in particular tumour-necrosis factor-a, are a major driver of pathogenesis.
We previously developed a dual-layer electrospun mucoadhesive patch with high patient acceptability and a long in vivo residence time (Colley, HE et al., Biomaterials. 178:134-146 (2018)). We have incorporated small molecules drugs such as corticosteroids and anaesthetics into these patches and have demonstrated their release and activity (Clitherow KH, et al., Mol Pharm 16:3948-56 (2019)). Recently, we showed that small proteins and peptides can also be incorporated whilst retaining their biological activity (Edmans JG. et al., Mat Sci Eng 112:110917 (2020)). Here, we investigate the inclusion and delivery of therapeutic anti-TNF-α F(ab) antibody fragments for the treatment of oral inflammatory disease.
Methodology and results: Biotinylated F(ab) fragments was incorporated into an electrospun mucoadhesive membrane and were found to be retained within the fibres in aggregates when visualised by confocal microscopy. These F(ab) were rapidly eluted from the patch without loss of antigen binding activity (97 ± 5% released within 3h). Neutralising anti-TNF-α F(ab) fragments were generated from whole IgG by papain cleavage, as confirmed by SDS-PAGE, then incorporated into patches. Antibody-containing patches were found to have TNF-α neutralising activity, as shown by the suppression of TNF-α-mediated CXCL8 release from oral keratinocyte grown as monolayer cultures. Patches applied to lipopolysaccharide stimulated immune-competent oral mucosal ulcer equivalents that contained primary macrophages, led to a statistically significant reduction in the levels of biologically active TNF-α, suggesting successful delivery of a therapeutically relevant dose. Moreover, inhibition of TNF-α by patch-released anti-TNF-α F(ab) fragments also resulted in a significant decrease in the levels of T-cell chemokines produced, indicating that patch-delivered neutralising antibody therapy impacts on downstream immunological events such as leukocyte recruitment that will further prevent pathogenesis.
Conclusion: Electrospun oromucosal patches can deliver active biologics such as therapeutic antibodies topically to diseased oral mucosal sites in vitro. These patches have the potential to change the way these debilitating oral diseases are treated in the future."
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