Journal: Research

Article Title: A pH-Responsive Polysaccharide Hydrogel Modulates Oxygen Delivery and Immunometabolic Remodeling for Diabetic Bone Regeneration

doi: 10.34133/research.1428

Figure Lengend Snippet: GOZ6 improves mitochondrial function in BMSCs and promotes osteogenic differentiation. (A) Representative fluorescence images of JC-1 in BMSCs under different hydrogel treatments. Scale bar, 200 μm. (B) JC-1 flow cytometric analysis of BMSCs in different groups. (C) Representative fluorescence images of ROS staining in BMSCs under different hydrogel treatments. (D) Representative images of alkaline phosphatase (ALP) staining and Alizarin Red S (ARS) staining. Scale bar, 100 μm. (E) Quantitative analysis of ALP activity and mineralized nodule formation. (F) Immunofluorescent staining of BMSCs cultured for 3 d: green (COL1), red (phalloidin), and blue (DAPI). Scale bar, 200 μm. (G) Quantitative analysis of COL1 fluorescence intensity. (H to J) RT-qPCR analysis of osteogenesis-related gene expression ( Alpl, Runx2, and Col1a1 ) in BMSCs. Data are presented as the mean ± SD ( n = 5). Compared with the GO group: * P < 0.05, ** P < 0.01, *** P < 0.001, ns: not significant ( P > 0.05). Compared with the GOZ6 group: # P < 0.05, ## P < 0.01, ### P < 0.001, ns: not significant ( P > 0.05).

Article Snippet: Mitochondrial membrane potential was evaluated using a JC-1 fluorescent probe (Beyotime, Haimen, China).

Techniques: Fluorescence, Analysis, Staining, Activity Assay, Cell Culture, Quantitative RT-PCR, Gene Expression

Journal: Advanced Science

Article Title: Carrier‐Free Berberine/Nitidine Chloride Self‐Assembled Nanoparticles Induce Ferroptosis to Overcome Bortezomib Resistance in Multiple Myeloma

doi: 10.1002/advs.77174

Figure Lengend Snippet: Berberine (BBR)/nitidine chloride (NC) self‐assembled nanoparticles (BBR/NC‐SAPs) display potent and selective anti‐myeloma activity by inhibiting proliferation, clonogenicity, DNA synthesis, and inducing mitochondrial dysfunction. (A) Multiple myeloma cells (sensitive: 8226, KMS‐11; bortezomib‐resistant: 8226‐BTZR, KMS‐11‐BTZR) and normal B cells (GM12878) were treated for 48 h with a concentration gradient (0, 2, 4, 6, 8, and 10 µg/mL) of free BBR and NC combination (1:1). (B) Multiple myeloma cells (sensitive: 8226, KMS‐11; bortezomib‐resistant: 8226‐BTZR, KMS‐11‐BTZR) and normal B cells (GM12878) were treated for 48 h with a concentration gradient (0, 2, 4, 6, 8, and 10 µg/mL) of BBR/NC‐SAPs. (C) Cells were treated with the indicated concentrations (0, 2, 4, 6, 8, and 10 µg/mL) of BBR/NC‐SAPs. Colony formation assay demonstrating the potent, dose‐dependent inhibition of clonogenic survival. (D) Cells were treated with the indicated concentrations (0, 2, 4, 6, 8, and 10 µg/mL) of BBR/NC‐SAPs. Analysis of DNA synthesis by EdU incorporation assay utilizing flow cytometry. The percentage of EdU‐positive (green) cells decreased in a dose‐dependent manner upon treatment. (E) Assessment of mitochondrial membrane potential by JC‐1 staining employing flow cytometry. Treatment with BBR/NC‐SAPs induced a dose‐dependent loss of mitochondrial membrane potential. Data are presented as the mean ± SD from three independent experiments. Statistical significance was determined by a one‐way ANOVA followed by Dunnett's multiple comparison test versus the control group. * p < 0.05, ** p < 0.01.

Article Snippet: The mitochondrial membrane potential (ΔΨm) was assessed utilizing the fluorescent probe JC‐1 (MCE, USA) according to the manufacturer's protocol.

Techniques: Activity Assay, DNA Synthesis, Concentration Assay, Colony Assay, Inhibition, Flow Cytometry, Membrane, Staining, Comparison, Control

Journal: Advanced Science

Article Title: Carrier‐Free Berberine/Nitidine Chloride Self‐Assembled Nanoparticles Induce Ferroptosis to Overcome Bortezomib Resistance in Multiple Myeloma

doi: 10.1002/advs.77174

Figure Lengend Snippet: NR2F2 is essential for berberine (BBR)/nitidine chloride (NC) self‐assembled nanoparticles (BBR/NC‐SAPs)‐induced ferroptosis in multiple myeloma (MM) cells. (A) Cytotoxicity assays showing that NR2F2‐knockout cells locked the survival threshold and abolished the dosage‐dependent cytotoxicity of BBR/NC‐SAPs as compared to the control. (B) Colony formation assays revealing that NR2F2 knockout preserves clonogenic potential after BBR/NC‐SAPs treatment. (C) Analysis of DNA synthesis by EdU incorporation assay utilizing flow cytometry, demonstrating that NR2F2 knockout preserves cell proliferation potential after BBR/NC‐SAPs treatment. (D) Assessment of mitochondrial membrane potential (ΔΨm) via JC‐1 or similar dye, denoting that NR2F2 deletion prevents membrane depolarization triggered by BBR/NC‐SAPs. (E) Quantification of intracellular Fe 2+ showing blocked iron accumulation in NR2F2‐deficient cells upon BBR/NC‐SAPs treatment. (F) Lipid ROS levels measured by flow cytometry demonstrating that BBR/NC‐SAPs fail to elevate lipid peroxidation in NR2F2‐knockout cells. Ferroptosis‐related protein levels measured by western blot showing that BBR/NC‐SAPs fail to elevate lipid peroxidation in NR2F2‐knockout cells (G, 8226 and 8226‐BTZR; H, KMS‐11 and KMS‐11‐BTZR).

Article Snippet: The mitochondrial membrane potential (ΔΨm) was assessed utilizing the fluorescent probe JC‐1 (MCE, USA) according to the manufacturer's protocol.

Techniques: Knock-Out, Control, DNA Synthesis, Flow Cytometry, Membrane, Western Blot

Journal: Bioactive Materials

Article Title: Injectable microgels carrying engineered biomimetic nanoparticles for osteoarthritis therapy via dual-targeted senescent chondrocyte clearance and endogenous repair promotion

doi: 10.1016/j.bioactmat.2025.11.038

Figure Lengend Snippet: Preparation and characterization of ANCM@SHM. (a) Schematic diagram of NCM preparation. (b) Representative TEM images of native CM and NCM. Bar: 50 nm. (c–d) Zeta potential (n = 3), particle size (n = 3), and PDI (n = 3) of native CM and NCM measured by DLS. (e) Representative TEM images of Lipo, A-lipo, and ANCM. Bar: 50 nm. (f–g) Zeta potential (n = 3), particle size (n = 3), and PDI (n = 3) of Lipo, A-lipo, and ANCM measured by DLS. (h) Representative confocal fluorescence micrographs showing red fluorescently labeled A-lipo and green fluorescently labeled NCM fusing into gold fluorescent ANCM. Bar: 10 μm. (i) Quantification of NKG2D on the surface of CM, NCM, and ANCM nanoparticles by flow cytometry. (j–k) Encapsulation efficiency (EE) (n = 3) and loading efficiency (LE) (n = 3) of ABT263. (l) Representative SEM image of ANCM@SHM. Bar: 100 μm. (m) Representative fluorescence image of PKH67-ANCM@SHM captured by confocal microscope. Bar: 100 μm.

Article Snippet: Subsequently, cells were incubated in the dark at room temperature for 20 min. Mitochondrial membrane potential was assessed employing the JC-1 fluorescent probe (Beyotime, China) following manufacturer's protocols.

Techniques: Zeta Potential Analyzer, Fluorescence, Labeling, Flow Cytometry, Encapsulation, Microscopy