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郝颖, 徐润壕, 陈茂, 等. 共载雷帕霉素和光克洛的纳米药物复合可溶解微针治疗鲜红斑痣的实验研究[J]. koko体育app 学报(医学版), 2024, 55(2): 433-440. DOI:
引用本文: 郝颖, 徐润壕, 陈茂, 等. 共载雷帕霉素和光克洛的纳米药物复合可溶解微针治疗鲜红斑痣的实验研究[J]. koko体育app 学报(医学版), 2024, 55(2): 433-440. DOI:
HAO Ying, XU Runhao, CHEN Mao, et al. Rapamycin and HPPH Co-Loaded Nanodrug Delivered via Dissolvable Microneedles to Treat Port-Wine Stains[J]. Journal of Sichuan University (Medical Sciences), 2024, 55(2): 433-440. DOI:
Citation: ෴ HAO Ying, XU Runhao, CHEN Mao, et al. Rapamycin and HPPH Co-Loaded Nanodrug Delivered via Dissolvable Microneedles to Treat Port-Wine Stains[J]. Journal of Sichuan University (Medical Sciences), 2024, 55(2): 433-440. DOI:

共载雷帕霉素和光克洛的纳米药物复合可溶解微针治疗鲜红斑痣的实验研究

Rapamycin and HPPH Co-Loaded Nanodrug Delivered via Dissolvable Microneedles to Treat Port-Wine Stains

  • 摘要:
    目的 鲜红斑痣是一种先天性毛细血管畸形的皮肤疾病,本研究结合鲜红斑痣的生物学特性和微针经皮传输药物的优势,拟构建共载抗血管生成药物雷帕霉素(rapamycin, RPM)和光敏剂光克洛(photochlor, HPPH)的纳米药物复合可溶解微针(RPM-HPPH NP@HA MN)实现抗血管生成治疗与光动力协同治疗鲜红斑痣。
    方法 首先通过乳化溶剂挥发法制备装载RPM和HPPH的纳米药物(RPM and HPPH co-loaded nanoparticles, RPM-HPPH NP),在660 nm激光照射下考察其活性氧(reactive oxygen species, ROS)产生能力,以小鼠血管内皮瘤细胞系EOMA为研究对象,通过荧光显微镜和流式细胞仪检测细胞摄取行为,MTT实验法检测在有、无660 nm激光照射下,RPM-HPPH NP对EOMA的细胞毒性作用(游离RPM为对照)。之后通过模板法将纳米药物与透明质酸(hyaluronic acid, HA)可溶解微针体系复合得到RPM-HPPH NP@HA MN,通过扫描电镜和电子万能试验机考察其形貌特征和力学性能,通过台盼蓝染色和HE染色实验评价RPM-HPPH NP@HA MN对裸鼠皮肤的刺入能力。
    结果 RPM-HPPH NP粒径150 nm,在激光的照射下可产生大量的ROS。在细胞水平,RPM-HPPH NP可被EOMA摄取,且呈时间依赖性。无论是否光照,纳米药物RPM-HPPH NP的细胞毒性强于游离药物RPM;在光照条件下,RPM-HPPH NP展现出更强的细胞毒性作用,差异有统计学意义(P<0.05)。RPM-HPPH NP@HA MN针尖高度是600 µm,单针力学性能为0.75048 N。台盼蓝染色和HE染色实验表明微针按压可使皮肤表面产生孔道并穿透角质层。
    结论 RPM-HPPH NP@HA MN可将RPM-HPPH NP经皮传输至病灶组织,实现抗血管生成治疗与光动力协同治疗鲜红斑痣,为纳米药物复合微针体系的构建及鲜红斑痣的治疗研究提供新策略。
     
    Abstract:
    Objective Port-wine stains are a kind of dermatological disease of congenital capillary malformation. Based on the biological characteristics of port-wine stains and the advantages of microneedle transdermal administration, we intend to construct a nanodrug co-loaded with rapamycin (RPM), an anti-angiogenesis drug, and photochlor (HPPH), a photosensitizer, and integrate the nanodrug with dissolvable microneedles (MN) to achieve anti-angiogenesis and photodynamic combination therapy for port-wine stains.
    Methods  First, RPM and HPPH co-loaded nanoparticles (RPM-HPPH NP) were prepared by the emulsification solvent-volatilization method, and its ability to generate reactive oxygen species (ROS) was investigated under 660 nm laser irradiation. Mouse hemangioendothelioma endothelial cells (EOMA) were used as the subjects of the study. The cellular uptake behaviors were examined by fluorescence microscopy and flow cytometry. The cytotoxicity effects of RPM-HPPH NP with or without 660 nm laser irradiation on EOMA cells were examined by MTT assays (with free RPM serving as the control). Then, hyaluronic acid (HA) dissolvable microneedles loaded with RPM-HPPH NP (RPM-HPPH NP@HA MN) were obtained by compounding the nanodrug with HA dissolvable microneedle system through the molding method. The morphological characteristics and mechanical properties of RPM-HPPH NP@HA MN were investigated by scanning electron microscope and electronic universal testing machine. The penetration ability of RPM-HPPH NP@HA MN on the skin of nude mice was evaluated by trypan blue staining and H&E staining experiment.
    Results The RPM-HPPH NP prepared in the study had a particle size of 150 nm and generated large amounts of ROS under laser irradiation. At the cellular level, RPM-HPPH NP was taken up by EOMA cells in a time-dependent manner. The cytotoxicity of RPM-HPPH NP was higher than that of free RPM with or without laser irradiation. Under laser irradiation, RPM-HPPH NP exhibited stronger cytotoxic effects and the difference was statistically significant (P<0.05). The height of the needle tip of RPM-HPPH NP@HA MN was 600 µm and the mechanical property of a single needle was 0.75048 N. Trypan blue staining and HE staining showed that pressing on the microneedles could produce pores on the skin surface and penetration of the stratum corneum.
    Conclusion RPM-HPPH NP@HA MN can deliver RPM-HPPH NP percutaneously to the lesion tissue and realize the synergistic treatment of port-wine stains with anti-angiogenic therapy and photodynamic therapy, providing a new strategy for the construction of nanodrug-loaded microneedle delivery system and the clinical treatment of port-wine stains.
     
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