Department of Dermatology and Wound Healing Centre, Copenhagen University Hospital, Bispebjerg and Frederiksberg, Copenhagen, 2400, Denmark.
Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, 1870, Denmark.
Lasers Surg Med. 2021 Jan;53(1):154-161. doi: 10.1002/lsm.23322. Epub 2020 Sep 30.
PD-L1 is a tumor ligand that binds to the PD-1 receptor on immune cells, thereby inhibiting the antitumor immune response. The antibody nivolumab is a PD-1 inhibitor, Food and Drug Administration approved for systemic treatment of several aggressive cancer types. Topically applied nivolumab may hold potential as a future strategy to treat keratinocyte cancer, but its molecular properties preclude unassisted topical uptake. The aim of this study was to investigate uptake and biodistribution of topically delivered nivolumab, assisted by two physical enhancement techniques with different delivery kinetics; ablative fractional laser (AFL) and electronically controlled pneumatic injection (EPI).
STUDY DESIGN/MATERIALS AND METHODS: In vitro porcine skin was exposed to CO AFL (20 mJ/mb, 5% density), followed by passive diffusion of nivolumab in a Franz cell (1 mg/ml, 18 hours, n = 6) or treated with EPI (4 bar) for immediate delivery of nivolumab (1 mg/ml, 10 minutes, n = 6). The resulting nivolumab skin concentrations were quantified by enzyme-linked immunosorbent assay (ELISA) at three skin depths (100, 500, and 1500 µm), comparing the uptake from assisted delivery with intact skin. Biodistribution of nivolumab in the skin for all interventions was visualized by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) and fluorescence microscopy.
Delivery of nivolumab by AFL-assisted passive diffusion and immediate EPI both resulted in significantly enhanced uptake of nivolumab in all skin depths compared with intact skin (P < 0.05). With AFL, nivolumab concentrations reached 86.3 µg/cm (100 µm), 105.8 µg/cm (500 µm), and 19.3 µg/cm (1500 µm), corresponding to 2-10% of the applied concentration, with the highest deposition in the mid dermis. Immediate EPI delivered 429.4 µg/cm (100 µm), 584.9 µg/cm (500 µm), and 295.9 µg/cm (1500 µm) into the skin, corresponding to 29-58% of the applied nivolumab concentration. From qualitative visualization of the biodistribution, it appeared that nivolumab distributed in a horizontal and continuous homogenous band in the upper and mid dermis through AFL-exposed skin, whereas EPI-delivery showed a deep focal deposition extending into the deep dermis.
AFL-assisted passive diffusion and immediate EPI-assisted delivery show the potential to deliver therapeutic antibodies locally. Future in vivo and pharmacokinetic studies would reveal the full potential for topical antibody delivery by energy-based devices. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.
PD-L1 是一种肿瘤配体，可与免疫细胞上的 PD-1 受体结合，从而抑制抗肿瘤免疫反应。抗体nivolumab 是一种 PD-1 抑制剂，已获美国食品药品监督管理局批准用于治疗几种侵袭性癌症类型的全身治疗。局部应用 nivolumab 可能是治疗角质形成细胞癌的一种潜在策略，但它的分子特性排除了非辅助性局部吸收。本研究旨在研究两种具有不同传递动力学的物理增强技术辅助下局部给予 nivolumab 的摄取和体内分布；消融性微点阵激光（ablative fractional laser，AFL）和电子控制气动注射（ electronically controlled pneumatic injection，EPI）。
研究设计/材料和方法：体外猪皮暴露于 CO AFL（20 mJ/mb，5%密度），然后在 Franz 细胞中进行 nivolumab 的被动扩散（1 mg/ml，18 小时，n = 6）或用 EPI（4 bar）进行即时给药（1 mg/ml，10 分钟，n = 6）。通过酶联免疫吸附试验（enzyme-linked immunosorbent assay，ELISA）在三个皮肤深度（100、500 和 1500 µm）处定量测定经皮给药后的 nivolumab 皮肤浓度，比较辅助给药与完整皮肤的摄取情况。采用激光烧蚀-电感耦合等离子体质谱（laser ablation-inductively coupled plasma-mass spectrometry，LA-ICP-MS）和荧光显微镜对所有干预措施的 nivolumab 在皮肤中的体内分布进行可视化。
AFL 辅助被动扩散和即刻 EPI 辅助给药均导致所有皮肤深度的 nivolumab 摄取明显增加，与完整皮肤相比（P < 0.05）。使用 AFL，nivolumab 浓度达到 86.3 µg/cm（100 µm）、105.8 µg/cm（500 µm）和 19.3 µg/cm（1500 µm），分别为应用浓度的 2-10%，沉积量最高的部位在中真皮。即刻 EPI 将 429.4 µg/cm（100 µm）、584.9 µg/cm（500 µm）和 295.9 µg/cm（1500 µm）输送到皮肤中，分别为应用 nivolumab 浓度的 29-58%。从体内分布的定性可视化来看，AFL 暴露的皮肤中，nivolumab 似乎呈水平连续均匀分布在真皮的浅层和中层，而 EPI 给药显示出向深层真皮延伸的深焦点沉积。
AFL 辅助被动扩散和即刻 EPI 辅助给药显示出局部递送治疗性抗体的潜力。未来的体内和药代动力学研究将揭示基于能量的设备局部抗体给药的全部潜力。激光外科学杂志。© 2020 威利父子公司
