载药近红外荧光颗粒示踪的干细胞球对放射性皮肤损伤修复作用
MSC spheroids tracking with ICG-drug-co-loaded NIR polymeric nanoparticles for the repair of radiation-induced skin injury
目的：研究载药纳米荧光颗粒对人脐带间充质干细胞球(MSCs)的近红外荧光成像和放射性皮肤损伤的修复作用。方法：通过复乳法制备担载近红外荧光素ICG和药物AMD3100的聚乙二醇聚乙丙交酯纳米颗粒(ICG-A-NPs)，将其与MSCs在成球条件下共培养，制备载药荧光颗粒示踪的间充质干细胞球(ICG-A-NP@MSC spheroids)，通过动态光散射、扫描电子显微术、透射电子显微术、紫外吸收和荧光光谱表征荧光颗粒的物化性质；采用细胞毒性、凋亡实验评价荧光颗粒的生物相容性；采用激光共聚焦显微术观察颗粒对干细胞的荧光标记能力；将示踪干细胞球与受照人皮肤成纤维细胞WS1共培养，通过克隆形成、凋亡实验评价示踪干细胞球对WS1细胞的修复效果。结果：所制聚合物荧光颗粒的粒径约为120-300 nm，浓度高至400 μg/mL时，也不影响干细胞的增殖和凋亡；内吞荧光颗粒后细胞骨架结构无显著变化，证明纳米颗粒生物相容性良好,且小动物活体成像仪中细胞数高于0.5×10<sup>6</sup>时可与背景信号明显区分。与对照组相比，载药荧光颗粒示踪的间充质干细胞球可提高受照WS1细胞的克隆形成能力，减少细胞凋亡率，促进放射性皮肤损伤修复。结论：示踪干细胞球具有良好的近红外荧光成像能力，可减轻放射性皮肤损伤的发生发展，有望成为一种示踪损伤救治过程的活细胞药物。
Objective: To investigate mesenchymal stem cell spheroids labeled with near-infrared polymeric nanoparticles for the repair ofradiation-induced skin injury. Methods: ICG-A-NPswere prepared by using monomethoxy polyethylene glycol-poly (lactic-co-glycolic acid) (mPEG-PLGA), indocyanine green (ICG) and Plerixafor (AMD3100) through the double emulsion method(w/o/w). Subsequently,mesenchymal stem cell spheroids (ICG-A-NP@MSC spheroids) was harversted by incubation of MSCs with various concentration of ICG-A-NPsunder spheroidizing conditions. Dynamic light scattering, scanning electron microscopy, transmission electron microscopy, and fluorescence spectroscopy were adopted to characterize the surface morphology, particle size and fluorescent spectra of nanoparticles. Moreover, the cytotoxicity of ICG-A-NPs was evaluated bycell apoptosis assaies.The fluorescent images of ICG-A-NP@MSC spheroids were observe by confocal laser scanning microscopy. And then, the ICG-A-NP@MSC spheroids were co-cultured with X-ray irradiated human skin fibroblast cells (WS1), and the apoptosis and colony formation of WS1 cells were evaluated correspondingly. Results: The ICG-A-NPs with round shapeare 120-300nm approximately. Even when the concentration of ICG-A-NPs is up to 400μg/mL, no distinguishable cytotoxicity and apoptosis of MSCs were observed.The cytoskeleton of MSC spheroids were clearly illustrated by the encapsulated ICG-A-NPs in fluorescence microscopic field. The fluorescent signals of ICG-A-NP@MSC spheroids with the cell number higher than 0.5×10<sup>6</sup> can be clearly distinguished from the backgroundin IVIS.Compared with the control group,ICG-A-NP@MSC spheroids can improve the clone formation ability of irradiated WS1 cells, reduce the cell apoptosis rate, resulting inthe promotion of the repair of radiation-induced damage.Conclusion: ICG-A-NP@MSC spheroidsexhibitedvisible near-infrared fluorescence imaging. Meanwhile, the spheroids can reduce the skincell apoptosis and ameliorate the colony formation capability, which is likely to benefit the imaging-guided repair of radiation-induced skin injury.
张舒羽、张琦、张慧琳
医药卫生理论基础医学生物科学研究方法、生物科学研究技术
纳米颗粒间充质干细胞细胞球荧光成像放射性皮肤损伤
Nanoparticlesmesenchymal stem cellsspheroidsfluorescent imagingradiation-induced skin injury
张舒羽,张琦,张慧琳.载药近红外荧光颗粒示踪的干细胞球对放射性皮肤损伤修复作用[EB/OL].(2024-05-20)[2025-08-11].http://www.paper.edu.cn/releasepaper/content/202405-115.点此复制
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