Journal: Journal of Nanobiotechnology

Article Title: Uterine metabolic disorder induced by silica nanoparticles: biodistribution and bioactivity revealed by labeling with FITC

doi: 10.1186/s12951-021-00810-x

Figure Lengend Snippet: Impact of SiNP internalization on the viability and biological function of the HTR8 cells. a Images showing the biodistribution of FITC-SiNP internalized by the HTR8 cells (×10 magnification). The nuclei were stained with Hoechst in blue, SiNP was conjugated with FITC in green, and the cytoskeleton was stained with Tubulin in red. b Images showing the suppressed proliferation of the HTR8 cells, where the HTR8 cells were treated with SiNP without FITC conjugation, the nuclei were stained with DAPI in blue, the HTR8 cells in the proliferation stage were stained with EdU in green (×10 magnification). c CCK8 assay showing the attenuated cell viability of the SiNP-exposed HTR8 cells. d Quantitative analysis of the proliferation of the HTR8 cells. e Confocal laser scanning images showing the apoptotic HTR8 cells, where the apoptotic cells were labeled with white dots (×10 magnification). f Quantitative analysis of apoptotic HTR8 cells. g Images showing the wounds scratched in the HTR8 cells. The attenuated wound healing ability was represented by the reduced area of the wound (×10 magnification). h Quantitative analysis of wound healing ability of the HTR8 cells. i Optical microscopy images showing tubes formed by the HTR8 cells (×4 magnification). j Quantitative analysis of the total tube length formed by the HTR8 cells. The differences were compared using one-way ANOVA followed by SNK post-hoc test. * P < 0.05

Article Snippet: According to the detection result of CCK8 assay, the median lethal dose (LD 50 ) of SiNP on trophoblast was determined as 100 µg/mL and was used to treat the HTR8 cells for 2 h. After rinsing the cells with PBS, the cytoskeleton was stained with Docetaxel for 30 min, and the nuclei were stained with Hoechst solution for 30 min. Then, the internalization of FITC- SiNP by the HTR8 cells was observed using ImageXpress Micro Confocal system (Molecular Devices, California, USA), and three absorbance spectra of 364 nm, 490 nm, and 652 nm were applied to visualize the nuclei, SiNPs, and cell skeleton.

Techniques: Staining, Conjugation Assay, CCK-8 Assay, Labeling, Microscopy

Journal: Journal of Nanobiotechnology

Article Title: Uterine metabolic disorder induced by silica nanoparticles: biodistribution and bioactivity revealed by labeling with FITC

doi: 10.1186/s12951-021-00810-x

Figure Lengend Snippet: High throughput sequencing and bioinformatics analysis of HTR8 cells with different interventions. a Boxplot showing the gene expression of HTR8 cells. Four samples of the HTR8 cells were exposed to SiNP at 100 µg/mL for 24 hours, and the other four samples were exposed to SiNP-free cell medium and were used as controls. Ctrl, control group; SiNPs, SiNP exposure group. b Density plot showing the gene expression of each sample. c Pearson correlation analysis between HTR8 cells with different interventions. The coefficients between different samples in the same group (red) were much higher than that in different groups (blue). d Principal component analysis of HTR8 cells with different interventions. All samples were clustered into two groups in accordance with their actual treatments. e Volcano plot showing the expression of genes in the HTR8 cells. Green, downregulation; red, upregulation; black, not significant. f Heatmap showing the dysregulated genes in SiNP-exposed HTR8 cells. The clustering analysis was conduced among different samples or dysregulated genes. Blue, downregulation; red, upregulation. g Topological network of the functional annotation and enrichment analysis based on the dysregulated genes. Different functional modules were labeled with different colors

Article Snippet: According to the detection result of CCK8 assay, the median lethal dose (LD 50 ) of SiNP on trophoblast was determined as 100 µg/mL and was used to treat the HTR8 cells for 2 h. After rinsing the cells with PBS, the cytoskeleton was stained with Docetaxel for 30 min, and the nuclei were stained with Hoechst solution for 30 min. Then, the internalization of FITC- SiNP by the HTR8 cells was observed using ImageXpress Micro Confocal system (Molecular Devices, California, USA), and three absorbance spectra of 364 nm, 490 nm, and 652 nm were applied to visualize the nuclei, SiNPs, and cell skeleton.

Techniques: Next-Generation Sequencing, Expressing, Functional Assay, Labeling

Journal: Journal of Nanobiotechnology

Article Title: Uterine metabolic disorder induced by silica nanoparticles: biodistribution and bioactivity revealed by labeling with FITC

doi: 10.1186/s12951-021-00810-x

Figure Lengend Snippet: Pathogenesis of SiNP-induced HTR8 cell apoptosis. The HTR8 cells were exposed to SiNP at 25, 50, 100, 200 µg/mL for 24 hours, and the cells exposed to SiNP-free medium were used as controls. a Expression of Caspase-3, ACOT1, SCD1, and CPT1A detected by Western blot in the HTR8 cells. b – e Quantitative analysis of Caspase-3, ACOT1, SCD1, and CPT1A by normalizing to GAPDH, where the expression of Caspase-3, ACOT1 and CPT1A was negatively correlated with SiNP, but SCD1 showed positive correlationship. f Immunofluorescence images showing the apoptotic and necrotic HTR8 cells induced by SiNP exposure (×10 magnification). The apoptotic HTR8 cells were stained in green with Annexin V-FITC, and the necrotic HTR8 cells were stained in red with propidium iodide. g Quantitative analysis of the apoptotic and necrotic HTR8 cells in different groups. h Schematic illustration of the pathogenesis of SiNP-induced HTR8 cell apoptosis. The differences between different treatment groups were compared using one-way ANOVA followed by SNK post-hoc test. * P < 0.05

Article Snippet: According to the detection result of CCK8 assay, the median lethal dose (LD 50 ) of SiNP on trophoblast was determined as 100 µg/mL and was used to treat the HTR8 cells for 2 h. After rinsing the cells with PBS, the cytoskeleton was stained with Docetaxel for 30 min, and the nuclei were stained with Hoechst solution for 30 min. Then, the internalization of FITC- SiNP by the HTR8 cells was observed using ImageXpress Micro Confocal system (Molecular Devices, California, USA), and three absorbance spectra of 364 nm, 490 nm, and 652 nm were applied to visualize the nuclei, SiNPs, and cell skeleton.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Journal of Nanobiotechnology

Article Title: Uterine metabolic disorder induced by silica nanoparticles: biodistribution and bioactivity revealed by labeling with FITC

doi: 10.1186/s12951-021-00810-x

Figure Lengend Snippet: Uterine expression of genes associated with HTR8 cell apoptosis in vitro. a Immunohistochemical images showing the expression of SCD1, CPT1A, ACOT2, and Caspase-3 in vivo (×20 magnification). The positive area was stained in brown. b – e Quantitative analysis and comparison of ACOT2, Caspase-3, CPT1A, and SCD1 between different observation time points. Comparing with the uterine expression of specific genes at 0 h, the expression of CPT1A, ACOT2, and Caspase-3 increased at 2, 4, 8, 16, 24 h time points, whereas SCD1 decreased. The differences were compared using one-way ANOVA followed by SNK post-hoc test. * P < 0.05

Article Snippet: According to the detection result of CCK8 assay, the median lethal dose (LD 50 ) of SiNP on trophoblast was determined as 100 µg/mL and was used to treat the HTR8 cells for 2 h. After rinsing the cells with PBS, the cytoskeleton was stained with Docetaxel for 30 min, and the nuclei were stained with Hoechst solution for 30 min. Then, the internalization of FITC- SiNP by the HTR8 cells was observed using ImageXpress Micro Confocal system (Molecular Devices, California, USA), and three absorbance spectra of 364 nm, 490 nm, and 652 nm were applied to visualize the nuclei, SiNPs, and cell skeleton.

Techniques: Expressing, In Vitro, Immunohistochemical staining, In Vivo, Staining