Abstract: Objective  To construct a dual-targeting nano-delivery system (ANG-2-CMLNPs) based on an epigallocatechin gallate (EGCG)-Zn supramolecular nanocarrier coated with a fusion membrane comprising angiopep-2-modified liposomes and glioma cell membranes for efficient loading and brain delivery of si-Survivin. Methods  Dynamic light scattering, transmission electron microscopy(TEM), agarose gel electrophoresis, fluorescence spectrophotometry, and flow cytometry were used to systematically characterize the particle size, Zeta potential, morphology, encapsulation efficiency, stability, and cellular targeting, uptake behavior of the nano-system. Models of U87 MG glioma cells and HCMEC/D3 endothelial cells were used to evaluate the system's blood-brain barrier（BBB） penetration capability, tumor cell targeting efficiency, and in vitro anti-proliferative activity. Results  ANG-2-CMLNPs with an average particle size of approximately (89.33±13.25) nm and a Zeta potential of(-11.17±0.70) mV were prepared. The optimal encapsulation efficiency［(98.98±1.01)%］ was attained at an EZ NP/siRNA mass ratio of 125∶1.  TEM confirmed a spherical shape with an intact membrane structure. Flow cytometry results indicated significant uptake of ANG-2-CMLNPs by HCMEC/D3 cells, demonstrating potential for BBB penetration; its uptake efficiency in U87 MG cells was significantly higher than in the control groups (P<0.001). When the mass ratio of liposomes to cell membrane was 1∶5, the system achieved optimal uptake balance. MTT results showed that ANG-2-CMLNPs inhibited U87 MG cell proliferation in a concentration-dependent manner, with cell viability decreasing to 28.4% at an siRNA concentration of 100 nmol/L. Conclusion  This study constructed a nano-drug delivery system with BBB-crossing and tumor-targeting capabilities, exhibiting good stability, high encapsulation efficiency, and significant targeting ability. The optimized membrane fusion ratio (1∶5) provided a key parameter for the synergistic realization of dual-targeting functions, offering a new strategy for RNAi therapy of glioma.

Key words: glioblastoma, angiopoietin-2, lipid fusion membrane, small interfering RNA, EGCG metal nanoparticles, biomimetic nanosystem