光敏剂包封加剧光动力疗法后细胞反应，聚合物胶束作为生物调节剂自主发挥作用
Encapsulation of photosensitizer worsen cell responses after photodynamic therapy protocol and polymer micelles act as biomodulators on their own
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发表日期:2024 Sep 30
作者:
Rachel Brival, Nathan Ghafari, Anne-Françoise Mingotaud, Isabelle Fourquaux, Véronique Gilard, Fabrice Collin, Patricia Vicendo, Stéphane Balayssac, Laure Gibot
DOI:
10.1016/j.ijpharm.2024.124589
摘要
光动力疗法（PDT）是一种临床应用的光化学治疗方式，广泛用于皮肤科、眼科和肿瘤学。以Pheo a为模型光敏剂，无论是游离态还是包封于聚乙烯醇-ε-壳聚糖（PEO-PCL）聚合物胶束中。嵌段共聚物胶束是水溶性生物相容性纳米容器，具有极大潜力用于递送疏水性药物。实验中也测试了空的PEO-PCL胶束。目标是进行体外研究，分析游离和包封Pheo a对人类结直肠肿瘤HCT-116细胞的反应，包括细胞结构、质膜交换线、线粒体功能和代谢紊乱。在校准的PDT方案中，包封增强了Pheo a的渗透（流式细胞术、共聚焦显微镜）和细胞死亡（Prestoblue检测），引起细胞形态（SEM）和细胞骨架（共聚焦）的大规模变化，线粒体功能障碍和完整性丧失（TEM），质膜上的离子快速大量流动（ICP-OES、离子色谱），以及代谢变化，包括氨基酸和胆碱衍生物水平的升高（1H NMR）。详细的研究揭示了包封Pheo-PDT的多方面效应，强调在理解治疗效果时应考虑光敏剂及其递送系统的重要性。研究还提出空纳米载体的更广泛影响问题，并鼓励更全面探索其生物学效应。
Abstract
Photodynamic therapy (PDT) is a photochemical therapeutic modality used clinically for dermatological, ophthalmological and oncological applications. Pheo a was used as a model photosensitizer, either in its free form or encapsulated within poly(ethylene oxide)-block-poly(ε-caprolactone) (PEO-PCL) polymer micelles. Block copolymer micelles are water-soluble biocompatible nanocontainers with great potential for delivering hydrophobic drugs. Empty PEO-PCL micelles were also tested throughout the experiments. The goal was to conduct an in vitro investigation into human colorectal tumor HCT-116 cellular responses induced by free and encapsulated Pheo a in terms of cell architecture, plasma membrane exchanges, mitochondrial function, and metabolic disturbances. In a calibrated PDT protocol, encapsulation enhanced Pheo a penetration (flow cytometry, confocal microscopy) and cell death (Prestoblue assay), causing massive changes to cell morphology (SEM) and cytoskeleton organization (confocal), mitochondrial dysfunction and loss of integrity (TEM), rapid and massive ion fluxes across the plasma membrane (ICP-OES, ion chromatography), and metabolic alterations, including increased levels of amino acids and choline derivatives (1H NMR). The detailed investigation provides insights into the multifaceted effects of encapsulated Pheo-PDT, emphasizing the importance of considering both the photosensitizer and its delivery system in understanding therapeutic outcomes. The study also raises questions as to the broader impact of empty nanovectors per se, and encourages a more comprehensive exploration of their biological effects.