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. 2020 Mar 13;12(3):260. doi: 10.3390/pharmaceutics12030260

Figure 3.

Figure 3

(A) Confocal microscopy images of human mesenchymal stem cells (hMSCs) after 30, 90 and 240 min of incubation at 37 °C with different concentration of HA/CS-RhB NPs (50–100 μg/20,000 cells). Red fluorescent denotes HA/CS NPs; nuclear blue fluorescence of DNA with Hoechst33258 dye; positive expression for cluster of differentiation-44 (CD44) on hMSCs membrane was detected by using anti-CD44 primary antibody and FITC labeled secondary antibody (green fluorescence), scale bar = 20 µm. (B) Detail of the endocytic vesicles formation: confocal image of hMSCs after 90 min incubation with HA/CS-RhB NPs (50 µg/20,000 hMSCs), a yellow line, crossing a vesicle, was drawn and the analysis of the red and green fluorescence intensities across this line was reported in graph I and II, respectively (scale bar = 10 µm). (C) Detail of the endocytic vesicles formation: confocal image of hMSCs after 90 min incubation with HA/CS-RhB NPs (100 µg/20,000 hMSCs), a yellow line, crossing a vesicle, was drawn and the analysis of the red and green fluorescence intensities across this line was reported in graph I and II, respectively (scale bar = 10 µm). (D) Effect of treatment time on red fluorescent intensity of the internalized HA/CS-RhB NPs (50–100 μg/20,000 cells). Results are presented as mean ± SD.**** p< 0.001. (E) Red fluorescent intensities of HA/CS-RhB NPs and CS-RhB/TPP NPs (100 μg/20,000 hMSCs) localized in the cell cytoplasm after 30, 90, 240 min of incubation. Results are presented as mean ± SD. ** p value < 0.01.