Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Antitumor effect of organometallic compounds in osteosarcoma cell lines. Cell viability (WST-1 assay) in 2D cultures measured after the treatment of 143B Par (A), OS833 Par (B), 143B CisPt-R (C), and OS833 CisPt-R (D) with increasing concentrations of free or encapsulated drugs for 72 h. The effect of equivalent amounts of empty nanoparticles in each cell line is shown. IC 50 values for each treatment are indicated. Error bars represent the standard deviation of three independent experiments. Asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon) (**** p < 0.0001; two-way ANOVA Tukey's test). (E and F) Analysis of DNA damage (formation of −H2AX foci) in 143B CisPt-R cells treated with the IC 75 of CisPt (8.4 µM), Ru3 (0.9 µM), LIP-Ru3 (0.8 µM), or the equivalent amount of empty nanoparticles (LIP-Empty) for 24 hours. Representative images of immunofluorescence staining (scale bar = 25 µm) (E) and quantification of the formation of gH2AX foci in >100 cells (F) are shown. Asterisks indicate statistically significant differences (** p < 0.01; *** p < 0.001; One-way ANOVA Tukey's test). (G) Cell cycle distribution of 143B CisPt-R cells treated with the IC 50 of CisPt (5 µM), Ru3 (0.5 µM), LIP-Ru3 (0.5 µM), or the equivalent amount of empty nanoparticles (LIP-Empty) for 48 hours. The normalized percentage of cells in each cell cycle phase (mean ± standard deviation of three independent experiments) is shown. Asterisks indicate statistically significant differences in G 0 /G 1 phase (blue) between LIP-empty control and Ru3-formulations and in the G 2 /M phase (pink) between LIP-empty and CisPt (**** p < 0.0001; two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: WST-1 Assay, Standard Deviation, Immunofluorescence, Staining, Control

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Cytotoxic effect of organometallic compounds in 3D parental spheroids. Spheroid viability (CTG 3D assay) was measured after the treatment of 143B Par (A and B) and OS833 Par (C and D) spheroids with increasing concentrations of free and encapsulated drugs for 96 h. The effect of equivalent amounts of empty nanoparticles is shown. Representative images of 143B Par (A) and OS833 Par (C) spheroids treated with increasing concentrations of each compound are shown (scale bars = 250 µm). Likewise, cell survival curves and IC50 values (µM) for each condition in 143B Par (B) and OS833 Par (D) spheroids are presented as the mean ± the standard deviation of at least eight biological replicates. Asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon) (**** p < 0.0001; two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: Standard Deviation

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Cytotoxic effect of organometallic compounds in 3D cisplatin-resistant spheroids. Spheroid viability (CTG 3D assay) was measured after the treatment of 143B CisPt-R (A and B) and OS833 CisPt-R (C and D) spheroids with increasing concentrations of free and encapsulated drugs for 96 h. The effect of equivalent amounts of empty nanoparticles is shown. Representative images of 143B CisPt-R (A) and OS833 CisPt-R (C) spheroids treated with increasing concentrations of each compound are shown (scale bars = 250 µm). Cell survival curves and IC50 values (µM) for each condition in 143B CisPt-R (B) and OS833 CisPt-R (D) are presented as the mean ± standard deviation of at least eight biological replicates. Asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon) (* p < 0.05; ** p < 0.01; **** p < 0.0001; two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: Standard Deviation

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Effect of free and nano-encapsulated Ru3 in CSC-enriched 3D osteosarcoma cell cultures. 143B Par cells were plated at low density in tumorsphere medium and left to form tumorspheres for 9 days before being treated for 96 h with increasing concentrations of Ru3, LIP-Ru3, and cisplatin. Treatments with the same amount of empty nanoparticles (LIP-Empty) were also included. Representative images (scale bars = 250 μm) (A), quantification of the number of tumorspheres remaining after the treatment (B), and cell viability (WST-1 assay) (C) are presented. Data is presented as the mean ± the standard deviation of independent replicates. In panel B, black asterisks indicate statistically significant differences between controls and the indicated conditions in each treatment. In panel C, asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon); (** p < 0.01; *** p < 0.001; one-way (panel B) or two-way (panel C) ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: WST-1 Assay, Standard Deviation

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: In vivo antitumor effect of free and nanoencapsulated metallic compounds. Established 143B CisPt-R xenografts were randomly assigned to six different groups ( n = 5 per group) and treated i.v. with vehicle (PBS, control), empty liposomes (LIP-Empty), free Ru3 (4 mg/kg, Ru3), ruthenium-loaded liposomes (4 mg/kg, LIP-Ru3), RAPTA-C (4 mg/kg), and cisplatin (4 mg/kg) once a week for 3 weeks (cumulative dose of 12 mg/kg). (A) Curves representing the mean relative tumor volume of 143B CisPt-R xenografts during the treatments. Drug efficacy expressed as the percentage of TGI is indicated. (B) Average tumor weight (left panel) and images of the tumors (right panel) at the end of the experiment (day 20 after the start of the treatment). (C) Change in the body weights of mice during the treatments. Error bars represent the SEM and asterisks indicate statistically significant differences between groups (* p < 0.05; **** p < 0.0001, two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: In Vivo, Control, Liposomes

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: Identification of active compounds and target prediction in YHD. (A) Venn diagram of the target of YHD and the target of osteosarcoma. (B – D) Gene Ontology (GO) enrichment analysis results. (E, F) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis results. (G) The component-target-pathway-disease network implicated in the mechanism of YHD in osteosarcoma treatment. The triangles represent osteosarcoma, the diamonds represent pathways, the circles represent key genes, and the squares represent the active ingredients of YHD. (H) Heatmap of molecular docking score. A binding energy heatmap with a bluer color indicates a more stable binding. (I) Molecular docking visualization between the active components of YHD and key targets.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Binding Assay

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD selectively inhibits osteosarcoma (OS) cells without affecting the viability or apoptosis of normal human cells. (A, B) CCK8 assay detected the effect of YHD on the viability of OS cells at 24 h and 48 h. (C, D) Colony formation assay detected the effect of YHD on the colony-forming ability of OS cells. (E – G) Flow cytometry was used to detect the effect of YHD on the cell cycle of OS cells. (H – J) Western blot analysis detected the effect of YHD on the levels of proliferation-related proteins in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Colony Assay, Flow Cytometry, Western Blot, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD can inhibit the migration and invasion of osteosarcoma (OS) cells, and promote their apoptosis. (A, B) Scratch healing assay showed that YHD inhibited the migration of OS cells. (C, D) Transwell assay showed that YHD inhibited the invasion of OS cells. (E – G) Western blot analysis detected the effect of YHD on the levels of proteins related to migration and invasion in OS cells. (H–K) Flow cytometry was used to detect the effect of YHD on apoptosis in OS cells. (L – N) Western blot analysis detected the effect of YHD on the levels of apoptosis-related proteins in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Migration, Transwell Assay, Western Blot, Flow Cytometry, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD induces osteosarcoma (OS) cell death by increasing ROS levels. (A) The effect of YHD on ROS levels in OS cells was detected using the DCFH probe method. (B) After N-acetylcysteine (NAC) treatment, the effect of YHD on ROS levels in OS cells was detected using the DCFH probe method. (C) After NAC treatment, CCK8 assay detected the effect of YHD on the viability of OS cells at 24 h and 48 h. (D, E) After NAC treatment, scratch healing assay showed that YHD inhibited the migration of OS cells. (F, G) After NAC treatment, Transwell assay showed that YHD inhibited the invasion of OS cells. (H, I) After NAC treatment, flow cytometry was used to detect the effect of YHD on the apoptosis of OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD can induce mitochondrial dysfunction in osteosarcoma (OS) cells. (A) Real-time quantitative PCR was used to measure mitochondrial DNA (mtDNA) levels. (B, C) Western blot analysis detected the effect of YHD on the expression levels of proteins related to mitochondrial biogenesis in OS cells. (D, E) JC-1 staining detected the effect of YHD on the mitochondrial membrane potential in OS cells. (F, G) MitoSOX staining detected the effect of YHD on mitochondrial ROS levels in OS cells. (H) Seahorse XFe24 analyzer measured the effect of YHD on the oxygen consumption rate (OCR) in OS cells. (I) The effect of YHD on ATP content in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, Membrane, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD exerts anti-tumor effects on osteosarcoma (OS) cells through the PI3K/AKT and p38 signaling pathways. (A) Principal component analysis revealed a clear distinction in gene expression profiles between the control and YHD groups. (B) Volcano plot identified 3495 differentially expressed genes in the YHD group. (C) Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. (D – G) Gene Set Enrichment Analysis (GSEA) of control and YHD groups. (H, I) Western blot analysis detected the effect of YHD on proteins related to the PI3K/AKT and MAPK pathways in OS cells. (J, K) After the addition of a PI3K activator and a P38 inhibitor, scratch healing assay showed that YHD inhibited the migration of OS cells. (L, M) After the addition of a PI3K activator and a P38 inhibitor, JC-1 staining detected the effect of YHD on the mitochondrial membrane potential in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Protein-Protein interactions, Gene Expression, Control, Western Blot, Migration, Staining, Membrane, Standard Deviation