School of Clinical Dentistry, 19 Claremont Crescent, University of Sheffield, Sheffield S10 2TA, UK; Department of Chemistry, Brook Hill, University of Sheffield, Sheffield S3 7HF, UK.
School of Clinical Dentistry, 19 Claremont Crescent, University of Sheffield, Sheffield S10 2TA, UK.
J Control Release. 2022 Oct;350:146-157. doi: 10.1016/j.jconrel.2022.08.016. Epub 2022 Aug 18.
Chronic ulcerative oral mucosal inflammatory diseases, including oral lichen planus and recurrent aphthous stomatitis, are painful and highly prevalent, yet lack effective clinical management. In recent years, systemic biologic therapies, including monoclonal antibodies that block the activity of cytokines, have been increasingly used to treat a range of immune-mediated inflammatory conditions such as rheumatoid arthritis and psoriasis. The ability to deliver similar therapeutic agents locally to the oral epithelium could radically alter treatment options for oral mucosal inflammatory diseases, where pro-inflammatory cytokines, in particular tumour-necrosis factor-α (TNFα), are major drivers of pathogenesis. To address this, an electrospun dual-layer mucoadhesive patch comprising medical-grade polymers was investigated for the delivery of F(ab) biologics to the oral mucosa. A fluorescent-labelled F(ab) was incorporated into mucoadhesive membranes using electrospinning with 97% v/v ethanol as a solvent. The F(ab) was detected within the fibres in aggregates when visualised by confocal microscopy. Biotinylated F(ab) was rapidly eluted from the patch (97 ± 5% released within 3 h) without loss of antigen-binding activity. Patches applied to oral epithelium models successfully delivered the F(ab), with fluorescent F(ab) observed within the tissue and 5.1 ± 1.5% cumulative transepithelial permeation reached after 9 h. Neutralising anti-TNFα F(ab) fragments were generated from whole IgG by papain cleavage, as confirmed by SDS-PAGE, then incorporated into patches. F(ab)-containing patches had TNFα neutralising activity, as shown by the suppression of TNFα-mediated CXCL8 release from oral keratinocytes cultured as monolayers. Patches were applied to lipopolysaccharide-stimulated immune-competent oral mucosal ulcer equivalents that contained primary macrophages. Anti-TNFα patch treatment led to reduced levels of active TNFα along with a reduction in the levels of disease-implicated T-cell chemokines (CCL3, CCL5, and CXCL10) to baseline concentrations. This is the first report of an effective device for the delivery of antibody-based biologics to the oral mucosa, enabling the future development of new therapeutic strategies to treat painful conditions.
慢性溃疡性口腔黏膜炎症性疾病，包括口腔扁平苔藓和复发性阿弗他口炎，疼痛剧烈且普遍存在，但缺乏有效的临床治疗方法。近年来，包括阻断细胞因子活性的单克隆抗体在内的全身性生物疗法已越来越多地用于治疗类风湿关节炎和银屑病等一系列免疫介导的炎症性疾病。能够将类似的治疗剂局部递送至口腔上皮，可能会彻底改变口腔黏膜炎症性疾病的治疗选择，其中促炎细胞因子，特别是肿瘤坏死因子-α（TNFα），是发病机制的主要驱动因素。为了解决这个问题，研究了一种由医用级聚合物组成的双层静电纺丝粘膜粘附贴片，用于将 F(ab)生物制剂递送至口腔黏膜。通过静电纺丝将荧光标记的 F(ab)用 97%v/v 乙醇作为溶剂掺入粘膜粘附膜中。共聚焦显微镜观察显示，纤维内的 F(ab)呈聚集状态。生物素化的 F(ab)从贴片中迅速洗脱（3 小时内释放 97±5%），且抗原结合活性无损失。贴片应用于口腔上皮模型，成功递递了 F(ab)，组织内观察到荧光 F(ab)，9 小时后达到 5.1±1.5%的累积跨上皮渗透。通过木瓜蛋白酶切割从全 IgG 生成中和抗 TNFα F(ab)片段，SDS-PAGE 确认，然后将其掺入贴片。含 F(ab)的贴片具有 TNFα 中和活性，这表现为抑制 TNFα 介导的单层培养的口腔角质形成细胞释放 CXCL8。贴片应用于含有原代巨噬细胞的脂多糖刺激的免疫活性口腔黏膜溃疡等效物。抗 TNFα 贴剂治疗可降低活性 TNFα 的水平，并使与疾病相关的 T 细胞趋化因子（CCL3、CCL5 和 CXCL10）的水平降低至基线浓度。这是将基于抗体的生物制剂递送至口腔黏膜的有效装置的首次报道，为治疗疼痛性疾病的新治疗策略的发展奠定了基础。
