没食子酸改性氧化石墨烯光热治疗微针的制备与评价 ^*

Preparation and evaluation of gallic acid-modified graphene oxide microneedles for photothermal therapy

摘要

目的 制备一种用于治疗老年黄褐斑的抑制黑色素分泌的光热治疗微针, 并对其性能及效果进行评价。方法 使用没食子酸 (GA) 改良氧化石墨烯 (GO) 结构, 制备 GAGO 纳米复合材料; 使用紫外—可见分光光度法和傅里叶变换红外光谱验证 GAGO 的合成情况; 使用马尔文粒度仪检测 GO、GAGO 在水溶液、PBS 和 RPMI 1640 培养基中的稳定性; 筛选壳聚糖、聚乙烯基吡咯烷酮 (PVP)、聚乙烯醇三种基质材料并确定微成型灌注方法。使用光学显微镜和扫描电子显微镜表征其形貌; 使用 Instronnstron 压缩—拉伸测试机测试微针的机械性能; 使用 SD 大鼠皮肤测试微针的皮肤插入性能; 通过 808 nm 近红外 (NIR) 激光器评估微针的光热转换性能; 使用 B16 黑色素瘤细胞中对 GAGO 光热治疗的体外抑制效果进行验证。结果 GA 成功负载于 GO, 且具有较好的稳定性和理化性质。成功构建了 GAGO-PVP 微针, 其形貌表征美观, 便于浇筑与脱模, 针尖完整无气泡, 通过机械性能测试, 失效力达到 3.66 N/针, 能刺穿皮肤且不引发炎症, 具备良好的光热转换性能。与对照组比较, GAGO 在 NIR 激发时, 能够显著提高 B16 黑色素瘤细胞活性, 降低酪氨酸酶活性并抑制黑色素分泌, 差异均具有统计学意义 (P<0.001)。结论 GAGO NIR 治疗能显著抑制 B16 黑色素瘤细胞活性和黑色素的分泌, GAGO-PVP 微针具有应用于黄褐斑治疗的潜力。

Abstract

Objective To prepare a photothermal therapeutic microneedle for the treatment of age-related chloasma that inhibits melanin secretion, and to evaluate its performance and efficacy. Methods GAGO nanocomposites were prepared by modifying the graphene oxide (GO) structure with gallic acid (GA); the synthesis of GAGO was verified using UV-Vis spectrophotometry and Fourier transform infrared spectroscopy; the stability of GO and GAGO in aqueous solution, PBS and RPMI 1640 medium was detected using a Malvern particle sizer; three matrix materials, chitosan, poly-vinyl pyrrolidone (PVP), and polyvinyl alcohol, were screened and the micromolding perfusion method was determined. Their topography was characterized using light microscopy and scanning electron microscopy; mechanical properties of microneedles were tested using an Instronnstron compression-tension tester; skin insertion properties of microneedles were tested using SD rat skin; photothermal conversion properties of microneedles were assessed by an 808 nm near-infrared (NIR) laser; and the in vitro inhibitory effect of GAGO photothermal therapy was verified using B16 melanoma cells. Results GA was successfully loaded on GO and had good stability and physicochemical properties. GAGO-PVP microneedles were successfully constructed, with beautiful appearance, easy pouring and demoulding, intact needle tips without bubbles, and through mechanical properties testing, the loss of efficacy reached 3.66 N/needle, which can penetrate the skin without causing inflammation, and has good photothermal conversion properties. Compared with the control group, GAGO could significantly increase the activity of B16 melanoma cells, decrease tyrosinase activity and inhibit melanin secretion during NIR challenge, and the differences were statistically significant (P<0.001). Conclusion GAGO NIR treatment significantly inhibited B16 melanoma cell viability and melanin secretion, and GAGO-PVP microneedles have the potential to be applied in melasma treatment.

关键词

氧化石墨烯 / 没食子酸 / 黄褐斑 / 微针 / 光热疗法

Key words

Graphene oxide / Gallic acid / Melasma / Microneedle / Photothermal therapy

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基金资助

^*中央本级重大增减支项目 “名贵中药资源可持续利用能力建设项目”(2060302)

国家自然科学基金项目(82304733)

中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-018)

吉林省科技发展计划项目(20230401102YY)