初步构建与评估递送柚皮素的RGD外泌体

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Abstract Objective： To prepare biosynthetic nanocarriers to deliver naringenin (Ng) and to investigate its in vitro tumor-suppressive effect on breast cancer. Methods： 293F cells secreting Arg-Gly-Asp-prostaglandin F2 receptor negative regulatorgreen fluorescent protein exosomes (RGD-P-G-Exo) were constructed by transfection using Arg-Gly-Asp-prostaglandin F2 receptor negative regulator-green fluorescent protein (RGD-P-G-Exo) plasmid. The nanomedicine (RGD-P-G-Exo@Ng) was prepared by collecting RGD-P-G-Exo in the supernatant by ultracentrifugation, adding Ng, and inducing the RGD-P-G-Exo to encapsulate the Ng by sonication in a water bath. 293F cells transfected with RGD-P-G were characterized using fluorescence microscopy. The morphology, particle size, markers and green fluorescent protein (GFP) expression of RGD-P-G-Exo@Ng were characterized by transmission electron microscopy, nanoparticle tracking analysis technique and protein immunoblotting. The encapsulation rate and drug loading capacity were assessed by UV spectrophotometry. Phagocytic uptake of the nanodrugs was assessed by cellular uptake assay. The killing ability of Exo@Ng and RGD-P-G-Exo@Ng on MDA-MB-231 cells was detected by CCK-8 assay. Results： Electron microscopy revealed a cuplike structure of RGD-P-G-Exo, with a particle size of 147.0 nm according to nanoparticle tracking analysis. Immunoblotting confirmed the surface expression of exosome markers (CD9, CD63) and GFP on RGD-P-G-Exo, validating the successful construction of RGD-P-G-Exo. The encapsulation efficiency of RGD-P-G-Exo@Ng was (28.1±0.6)%, and the drug loading content was (6.0±0.1)%. At high concentrations, Exo and RGD-P-G-Exo had no significant effect on the viability of MDA-MB-231 cells with good biocompatibility. In the cellular uptake assay, the RGD-P-G-Exo@Ng group had a stronger cellular uptake effect compared with the Exo@Ng group (P<0.05). In the pharmacodynamic assay, both Exo@Ng and RGD-P-G-Exo@Ng groups exhibited concentration-dependent cytotoxicity in MDA-MB-231 cells, in which the RGD-P-G-Exo@Ng group possessed a stronger cell-killing ability compared with the same concentration of Exo@Ng group (P<0.05). Conclusion： In this research, a nanoparticle for targeted delivery of Ng was initially synthesized, providing a new tool for the development of bio-nanotargeted tumor drug delivery systems.

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Received: 27 November 2023

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	FENG Qi1
	LIU Hongfeng2

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FENG Qi1,LIU Hongfeng2. Preliminary construction and evaluation of RGD exosomes for delivering naringin[J]. Journal of Jiangsu University(Medicine Edition), 2024, 34(05): 407-413.

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http://zzs.ujs.edu.cn/xbyxb/EN/ OR http://zzs.ujs.edu.cn/xbyxb/EN/Y2024/V34/I05/407

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