Title:Tumor Targeting with Apatinib-loaded Nanoparticles and
Sonodynamic Combined Therapy
Volume: 24
Issue: 5
Author(s): Xiao Han, Caifu Zhou, Xiaoling Luo*, Hongbing Pang, Chuangye Han, Libo Tang, Ziye Yang, Yingdan Nong and Chunmiao Lu
Affiliation:
- Department of Experimental Research, Guangxi Medical University Cancer Hospital, Nanning, China
Keywords:
ZIF-8, biomimetic nanocarrier, sonodynamic therapy, apatinib, targeted cancer therapy, Breast cancer, aCZM.
Abstract:
Introduction: This study implies the enhancement of apatinib killing effect in
4T1 tumor cells through constructing drug-loaded nanoparticles apatinib/Ce6@ZIF-
8@Membranes (aCZM) to enhance tumor therapeutic targeting and reduce toxic side
following sonodynamic therapy (SDT).
Methods: apatinib/Ce6@ZIF-8 (aCZ) were synthesized by in situ encapsulation, and
aCZM were constructed by encapsulating the nanoparticles with extracted breast cancer
4T1 cell membranes. aCZM were characterized and tested for the stability by electron
microscopy, and the membrane proteins on the nanoparticles’ surface were assessed
using SDS-PAGE gel electrophoresis. The cell viability of 4T1 cells following treatment
with aCZM was tested using cell counting kit-8 (CCK-8). The uptake of nanoparticles
was detected by laser confocal microscopy and flow cytometry, and the SDT-mediated
production of reactive oxygen species (ROS) was verified by singlet oxygen sensor
green (SOSG), electron spin resonance (ESR), and DCFH-DA fluorescent probes. The
CCK-8 assay and flow cytometry using Calcein/PI were used to assess the antitumoral
effect of aCZM nanoparticles under SDT. The biosafety of aCZM was further verified in
vitro and in vivo using the hemolysis assay, routine blood test and H&E staining of vital
organs in Balb/c mice.
Results: aCZM with an average particle size of about 210.26 nm were successfully
synthesized. The results of the SDS-PAGE gel electrophoresis experiment showed that
aCZM have a band similar to that of pure cell membrane proteins. The CCK-8 assay
demonstrated the absence of effects on cell viability at a low concentration range, and
the relative cell survival rate reached more than 95%. Laser confocal microscopy and
flow cytometry analysis showed that aCZM treated group has the strongest fluorescence
and the highest cellular uptake of nanoparticles. SOSG, ESR, and DCFH-DA fluorescent
probes all indicated that the aCZM + SDT treated group has the highest ROS production.
The CCK-8 assay also showed that when the ultrasound intensity was fixed at
0.5 W/cm2, the relative cell survival rates in the medium concentration group (10 μg/ml)
(5.54 ± 1.26%) and the high concentration group (20 μg/ml) (2.14 ± 1.63%) were significantly
lower than those in the low concentration group (5 μg/ml) (53.40 ± 4.25%).
Moreover, there was a concentration and intensity dependence associated with the cellkilling
effect. The mortality rate of the aCZM in the ultrasound group (44.95 ± 3.03%)
was significantly higher than that of the non-ultrasound (17.00 ± 2.26%) group and aCZ
+ SDT group (24.85 ± 3.08%) (P<0.0001). The live and dead cells’ staining (Calcein/PI)
also supported this result. Finally, in vitro hemolysis test at 4 and 24 hours showed that
the hemolysis rate of the highest concentration group was less than 1%. The blood
routine, biochemistry, and H&E staining results of major organs in Balb/c mice
undergoing nano-treatments showed no obvious functional abnormalities and tissue
damage in 30 days.
Conclusion: In this study, a multifunctional bionic drug delivery nanoparticles (aCZM)
system with good biosafety and compatibility in response to acoustic dynamics was
successfully constructed and characterized. This system enhanced apatinib killing effect
on tumor cells and reduced toxic side effects under SDT.
