Fig. 5: Deep penetration of CatNbots into the tumor.
From: A self-directed Trojanbot-enzymatic nanobot in neutrobot for active target therapy of glioblastoma
a Schematic diagram of the chemotaxis of CatNbots toward tumor tissue and the internalization by tumor cells. b Schematic of chemotactic motion of CatNbots along the concentration gradient of the H2O2. c Fluorescent images of the different chambers. d The corresponding fluorescence quantification values at different time points (n = 3). e In vitro penetration results in 3D tumor spheroids. The bright-field images and Z-stack CLSM images were taken from the top to the equatorial plane in 160 µm thickness of 3D cellular spheroids after treatment with GeNPs, EMV@GeNPs, and CatNbots for 4 h. f The corresponding fluorescent spectra and g the mean fluorescence intensity of the GL261 tumor spheroids treated with different samples (n = 3) (exact P value: GeNP vs. CatNbot P = 8.16414E−5). h Tumor spheroid morphology and TUNEL apoptosis assay of GL261 3D spheroids treated with various groups for 3 days. Spheroid slices, 160 µm thick, were subjected to the TUNEL assay, with positive cells identified using CLSM imaging. i Volume statistics of tumor spheroids before and after treatment across different experimental groups. j Cytotoxicity assessment of GeNPs, EMV@GeNPs, and CatNbots on 3D tumor spheroids (n = 3). k Cellular uptake CLSM images for different samples (Green: labeled with Cou6, Red: cell membranes stained with Dil, Blue: cell nuclei stained with Hoechst 33342). l The corresponding fluorescence semi-quantitative analysis (n = 3) (exact P values: GeNP vs. CatNbot P = 3.77E−7, EMV@GeNP vs. CatNbot P = 1.2024E−6). m Cytotoxicity of GeNPs, EMV@GeNPs, and CatNbots on GL261 cell lines in a 2D culture (n = 3). n Mechanism diagram of CatNbots regulating the TME. o Typical CLSM images of intracellular H2O2 (labeled with the H2O2 fluorescent probe, ROSGreenTM H2O2 Probe) in cancer cells after CatNbots treatment for 12 h (n = 3). p Relative H2O2 level in cancer cells after treatment with different samples for 12 h (n = 3) (exact P-value: 3 h Control vs. CatNbot P = 1.51132E−5, 3 h GeNP vs. CatNbot P = 3.32681E−5, 9 h Control vs. CatNbot P = 3.9146E−6, 9 h GeNP vs. CatNbot P = 2.219E−6, 9 h EMV@GeNP vs. CatNbot P = 3.617E−7, 12 h Control vs. CatNbot P = 3.18E−8, 12 h GeNP vs. CatNbot P = 2.9E−8, 12 h EMV@GeNP vs. CatNbot P = 1.72E−8). Data are presented as mean ± SD of three independent experiments (d, g, j, l, m, p). P-values are determined by one-way ANOVA and Tukey multiple comparisons tests in (g, l, p). Source data are provided as a Source Data file.
