超声、微球和叶酸修饰壳聚糖纳米粒介导的基因传递研究

Title:Research of Gene Delivery Mediated by Ultrasound, Microbubble and Folate-modified Chitosan Nanoparticles

Volume: 18 Issue: 6

关键词: 壳聚糖，超声波，基因治疗，叶酸，靶向，纳米颗粒。

摘要: Objective: To study transfection efficiency of folate-modified chitosan (FACS) nanoparticles as a non-viral vector delivering pEGFP-C3plasmid (FA-CS/P) to 293T cells with or without the combination of ultrasound and microbubble.

Method: pEGFP-C3 was used as a reporter gene and FA-CS nanoparticles, which were prepared by complex coagulation method, were used as biological carriers. Transfection efficiency to 293T cells mediated by FA-CS/P nanoparticles, ultrasound (US) and microbubble (MB) was assessed by fluorescence microscopy and flow cytometry.

Result: FA-CS/P nanoparticles have a particle size of 355.1 nm and zeta potential of 10.4 mV. Significant green fluorescence could be observed in CS/P group, FA-CS/P group, US+MB/P group, US+FA-CS/P group, Liposome 2000 (L) group under an inverted fluorescence microscope, while for US+MB+FA-CS/P group, only scattered fluorescence was observed. Result of flow cytometry showed that the transfection rate of US+MB+FA-CS/P group was (2.0±0.2)%, which was significantly lower than other groups (P<0.05). CCK-8 experiments showed that cell vitality of US+MB+FA-CS/P was (64.1±4.6)%, which was also lower than other groups (P<0.05).

Conclusion: In this study, FA-CS was successfully synthesized. FA-CS could combine with pEGFP-C3 effectively forming nanoparticles with nanoparticle size, well dispersion, high encapsulation efficiency and no significant toxicity to cells. The application of ultrasound increased the transfection rate of FA-CS/P. However, while being exposed to ultrasound and microbubble, the transfection rate of FA-CS/P decreased obviously, may indicate that there was no synergistic effect for gene transfection by the combination of ultrasound, folate modified chitosan and microbubbles.

Cite this article as:

Research of Gene Delivery Mediated by Ultrasound, Microbubble and Folate-modified Chitosan Nanoparticles, Current Molecular Medicine 2018; 18 (6) . https://dx.doi.org/10.2174/1566524018666181109121509