Effects of Photodynamic Therapy Using 5 -Aminolevulinic Acid (ALA) Loaded Acrylic Nanoparticles (ANPs) on HaCaT Cells

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-05-31 DOI:10.2147/ccid.s458525

Kang Ge, Yilu Zhao, Xiao Liu, Ruzhi Zhang

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引用次数: 0

Abstract

Objective: ALA-PDT (5-aminolevulinic acid photodynamic therapy) is a central modality in the treatment of skin diseases. Increasing the bioavailability of ALA remains a critical issue. With this in mind, our study explores a novel route of ALA delivery by loading acrylic nanoparticles (ANPs).
Methods: ALA-ANPs were synthesized by emulsion polymerisation and characterised by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The effects of ALA-ANPs on HaCaT cell line were evaluated, including characteristics, morphological changes, protoporphyrin IX (PpIX) fluorescence kinetics, reactive oxygen species (ROS) levels, mitochondrial membrane potential and ki67 expression in these cells.
Results: The ANPs had uniform sizes, smooth surfaces and excellent light transmittance, with diameters of 150– 200 nm. In contrast, the ALA - ANPs had uneven surfaces and poor light transmittance, with diameters of 220– 250 nm. During 12 hours of co-incubation of HaCaT cells with ALA, the intracellular accumulation of PpIX increased over time. Notably, after 6 hours of incubation, PpIX levels induced by 1.81 mg/mL ALA-ANPs exceeded those induced by 1.0 mM ALA (p < 0.01). CCK-8 results showed a positive correlation between PDT-induced inhibition of HaCaT cell proliferation and ALA concentration when ALA concentration remained below 2.0 mM. Compared to the 1.0 mM ALA group, the 1.81 mg/mL ALA-ANPs group showed decreased mitochondrial membrane potential, ki67 immunofluorescence intensity and cell proliferation. In contrast, ROS levels were significantly increased in the 1.81 mg/mL ALA-ANPs group (p < 0.01).
Conclusion: Loading ANPs provide improved stability and potency for ALA. The ALA-ANPs-PDT approach has superior inhibitory effects on HaCaT proliferation in vitro.

Keywords: 5-aminophenolvalic acid, ALA, acrylate nanoporous, photodynamic therapy

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使用 5 -氨基乙酰丙酸 (ALA) 负载丙烯酸纳米颗粒 (ANPs) 进行光动力疗法对 HaCaT 细胞的影响

目的：ALA-PDT（5-氨基乙酰丙酸光动力疗法）是治疗皮肤病的核心方法。提高 ALA 的生物利用率仍是一个关键问题。有鉴于此，我们的研究探索了一种通过负载丙烯酸纳米颗粒（ANPs）输送 ALA 的新途径：通过乳液聚合合成了 ALA-ANPs，并用扫描电子显微镜（SEM）、透射电子显微镜（TEM）和纳米粒子跟踪分析（NTA）对其进行了表征。评估了 ALA-ANPs 对 HaCaT 细胞系的影响，包括这些细胞的特征、形态变化、原卟啉 IX（PpIX）荧光动力学、活性氧（ROS）水平、线粒体膜电位和 ki67 表达：ANPs大小均匀，表面光滑，透光性极佳，直径为150-200纳米。相比之下，ALA - ANP 表面不平滑，透光性差，直径为 220- 250 nm。在 HaCaT 细胞与 ALA 共同培养 12 小时期间，PpIX 在细胞内的积累随着时间的推移而增加。值得注意的是，培养 6 小时后，1.81 mg/mL ALA-ANPs 诱导的 PpIX 水平超过了 1.0 mM ALA 诱导的水平（p < 0.01）。CCK-8 结果显示，当 ALA 浓度低于 2.0 mM 时，PDT 诱导的 HaCaT 细胞增殖抑制与 ALA 浓度呈正相关。与 1.0 mM ALA 组相比，1.81 mg/mL ALA-ANPs 组的线粒体膜电位、ki67 免疫荧光强度和细胞增殖均有所下降。相比之下，1.81 mg/mL ALA-ANPs 组的 ROS 水平明显升高（p < 0.01）：结论：负载 ANPs 提高了 ALA 的稳定性和效力。ALA-ANPs-PDT方法在体外对HaCaT增殖有较好的抑制作用：5-氨基苯戊酸 ALA 丙烯酸酯纳米孔光动力疗法

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.