Journal: Frontiers in Pharmacology

Article Title: Matrix type influences embedded patient-derived osteosarcoma organoid invasion and response to treatment

doi: 10.3389/fphar.2026.1831729

Figure Lengend Snippet: Workflow for patient-derived osteosarcoma (PDOS) organoid generation, embedding in photocrosslinkable ECMs, and analysis. (a) Method for generation of PDOS organoids. (b) On day 3 of culture, PDOS organoids were embedded in photocrosslinkable matrix precursors and crosslinked using visible light to prepare OS-matrix microenvironments. (c) Method for determination of organoid core and invasion area using brightfield images and ImageJ analysis. (d) Matrix-based response to doxorubicin (DOX) was used to evaluate the effect of matrix type and stiffness on OS response to chemotherapy.

Article Snippet: The viability of matrix-free organoids on day 7 and 14 of culture, and embedded organoids at the endpoint of DOX treatment (day 4 embedding), was determined using a fluorescein diacetate (FDA) (Thermo Fisher)/propidium iodide (PI) (Thermo Fisher) cell viability assay.

Techniques: Derivative Assay

Journal: Frontiers in Pharmacology

Article Title: Matrix type influences embedded patient-derived osteosarcoma organoid invasion and response to treatment

doi: 10.3389/fphar.2026.1831729

Figure Lengend Snippet: Doxorubicin (DOX) treatment of matrix-free and embedded PDOS organoids. (a) Fluorescein diacetate (FDA)-based staining of live PDOS organoids treated with mid-concentration DOX (0.1 µM) was conducted at endpoint of treatment (day 4 post-embedding). (b) Metabolic activity of matrix-free and embedded organoids determined on day 7 of culture (day 4 post-embedding). The relative metabolic activity of each sample was determined through normalization to respective group vehicle controls. Heatmap values represent mean. N = 1. n = 6. (c) IC 50 values for PDOS cultures was determined via [inhibitor] vs. response variable slope (four parameters) non-linear regression.

Article Snippet: The viability of matrix-free organoids on day 7 and 14 of culture, and embedded organoids at the endpoint of DOX treatment (day 4 embedding), was determined using a fluorescein diacetate (FDA) (Thermo Fisher)/propidium iodide (PI) (Thermo Fisher) cell viability assay.

Techniques: Staining, Concentration Assay, Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Matrix type influences embedded patient-derived osteosarcoma organoid invasion and response to treatment

doi: 10.3389/fphar.2026.1831729

Figure Lengend Snippet: Morphological response of PDOS organoids to treatment with increasing DOX concentration. (a) Brightfield microscopy of PDOS organoids treated with DOX demonstrates that core and invasive areas were reduced in all groups with increasing DOX concentration. (b) Quantification of spheroid core areas revealed that core size was slightly decreased with increasing DOX concentration for most matrix types. N = 1. n = 6. Error bars = STDEV. (c) Total organoid area quantification shows that groups with high invasion, such as soft LungMA and soft GelMA, undergo a decrease in total cell area with increasing DOX concentration. N = 1. n = 6. Error bars = STDEV. (d) Quantification of the percentage of core and invasive cell areas in relation to total cell coverage acts as an indicator for drug-based invasion inhibition. N = 1. n = 6. Error bars = STDEV.

Article Snippet: The viability of matrix-free organoids on day 7 and 14 of culture, and embedded organoids at the endpoint of DOX treatment (day 4 embedding), was determined using a fluorescein diacetate (FDA) (Thermo Fisher)/propidium iodide (PI) (Thermo Fisher) cell viability assay.

Techniques: Concentration Assay, Microscopy, Inhibition

Journal: STAR Protocols

Article Title: Protocol for preparing pH-sensitive Mg-DOX liposomes by remote loading for treatment of acidic tumors

doi: 10.1016/j.xpro.2026.104560

Figure Lengend Snippet: Preparation of MgAc 2 gradient liposomes and visual observation of the liposomes remote loaded with DOX (A) Platform illustrating the buffer exchange of MgAc 2 liposomes with 0.9% NaCl using a PD10 desalting column. (B) Photograph showing the color of MgAc 2 gradient liposomes, DOX·HCl and NaHCO 3 mixture stored at 25 °C for 5 min. (C) Photograph showing the color of the MgAc 2 gradient liposomes, DOX·HCl and NaHCO 3 mixture heated at 60 °C for 10 min.

Article Snippet: Doxorubicin Hydrochloride (DOX·HCl), USP grade , Shanghai yuanye Bio-Technology Co., Ltd. , CAS: 25316-40-9.

Techniques: Liposomes, Buffer Exchange

Journal: STAR Protocols

Article Title: Protocol for preparing pH-sensitive Mg-DOX liposomes by remote loading for treatment of acidic tumors

doi: 10.1016/j.xpro.2026.104560

Figure Lengend Snippet: Characterization of Mg-DOX liposomes for particle size and Zeta potential, phospholipid concentration, and DOX concentration (A) Size distribution of the Mg-DOX liposomes measured by DLS method. (B) Cryo-TEM micrograph of the Mg-DOX liposomes. Scale bar: 100 nm. (C) Standard curve of the Stewart’s assay of DSPC concentration. (D) Standard curve of the spectrophotometric determination of DOX·HCl concentration with the microplate reader.

Article Snippet: Doxorubicin Hydrochloride (DOX·HCl), USP grade , Shanghai yuanye Bio-Technology Co., Ltd. , CAS: 25316-40-9.

Techniques: Liposomes, Zeta Potential Analyzer, Concentration Assay

Journal: International Journal of Pharmaceutics: X

Article Title: GPC3 and CD133-targeted peptide dual modification enhances the therapeutic effect of doxorubicin carried by OMVs on hepatocellular carcinoma

doi: 10.1016/j.ijpx.2026.100510

Figure Lengend Snippet: Doxorubicin loading, release profile, and in vitro antitumor efficacy of dual-targeted OMVs. (A) Schematic illustration of the preparation of (GPC3 + CD133)ᵀ-OMVs@DOX. (B) Loading efficiency (LE) and drug content (DC) of (GPC3 + CD133)ᵀ-OMVs@DOX at different initial DOX concentrations. (C) In vitro DOX release profiles from (GPC3 + CD133)ᵀ-OMVs@DOX in PBS at pH 7.4 and pH 5.0. (D) Hydrodynamic diameter of (GPC3 + CD133)ᵀ-OMVs after DOX loading, as measured by DLS. (E) Cellular uptake of free DOX (red) and PKH67-labeled (green) (GPC3 + CD133)ᵀ-OMVs@DOX by Huh-7 cells after 6 h incubation. Scale bar: 100 μm. (F) Cell viability of Huh-7 cells treated with various concentrations of free DOX or (GPC3 + CD133)ᵀ-OMVs@DOX for 24 h, as determined by CCK-8 assay. (G) Cell viability of Huh-7 cells treated with different formulations at a concentration of 18.62 μM (IC50 of free DOX) for 24 h. Data are presented as mean ± SD ( n = 6). Statistical significance is denoted as follows: *p < 0.05, **p < 0.01, ***p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: DOX ( S17092 , Shanghai Yuanye, China) was loaded into OMVs using a sonication method.

Techniques: In Vitro, Labeling, Incubation, CCK-8 Assay, Concentration Assay

Journal: International Journal of Pharmaceutics: X

Article Title: GPC3 and CD133-targeted peptide dual modification enhances the therapeutic effect of doxorubicin carried by OMVs on hepatocellular carcinoma

doi: 10.1016/j.ijpx.2026.100510

Figure Lengend Snippet: In vivo antitumor efficacy and biodistribution of SsnB-pretreated and dual-targeted OMVs@DOX in a murine hepatoma model. (A) Schematic illustration of the experimental timeline and treatment groups ( n = 5). Mice received SsnB pretreatment followed by intravenous injection of: 1) PBS; 2) Blank OMVs; 3) Free DOX; 4) GPC3ᵀ-OMVs@DOX; 5) CD133ᵀ-OMVs@DOX; 6) (GPC3 + CD133)ᵀ-OMVs@DOX. (B) In vivo fluorescence imaging showing the biodistribution of Cy7.7 -labeled OMVs in tumor-bearing mice post-injection. (C) Ex vivo fluorescence imaging of dissected major organs and tumors at 24 h post-injection. # # #p < 0.001, # # p < 0.01, and # p < 0.05 vs. Oil/DMSO+SpC-OMVs group. (D) Tumor volume growth curves of mice during the treatment period. (E) Body weight curves of mice during the treatment period. (F) Representative photographs and weights of excised tumors at the endpoint of the study. (G) Immunohistochemical staining of Ki-67 in tumor tissues, showing proliferation index across treatment groups. Scale bar: 50 μm. Data are presented as mean ± SD (n = 5).Statistical significance is denoted as follows: #, compared to the control group (Oil/DMSO + SpC-OMVs); *, compared between treatment groups (#/* p < 0.05, ##/** p < 0.01, ###/*** p < 0.001,).

Article Snippet: DOX ( S17092 , Shanghai Yuanye, China) was loaded into OMVs using a sonication method.

Techniques: In Vivo, Injection, Fluorescence, Imaging, Labeling, Ex Vivo, Immunohistochemical staining, Staining, Control

Journal: International Journal of Pharmaceutics: X

Article Title: GPC3 and CD133-targeted peptide dual modification enhances the therapeutic effect of doxorubicin carried by OMVs on hepatocellular carcinoma

doi: 10.1016/j.ijpx.2026.100510

Figure Lengend Snippet: Biosafety evaluation of SsnB-pretreated and dual-targeted OMVs@DOX in tumor-bearing mice. (A) Serum levels of ALT and AST as markers of liver function. (B) Serum levels of creatinine (Cr) and Urea as markers of kidney function. (C) Representative hematoxylin and eosin (H&E)-stained sections of major organs (liver, spleen, kidneys, and heart) collected at the end of the treatment period. Scale bars: 100 μm. The H-E staining of the organs from other groups could be seen in Fig. S4. Arrows indicate atrophied myocardial cells. Data are presented as mean ± SD (n = 5). Statistical significance is denoted as follows: *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: DOX ( S17092 , Shanghai Yuanye, China) was loaded into OMVs using a sonication method.

Techniques: Staining