Journal: Bioactive Materials

Article Title: Neonatal muscle-derived extracellular vesicles containing miR-542-3p rejuvenate aged skeletal muscle via a functional microneedle patch

doi: 10.1016/j.bioactmat.2026.06.011

Figure Lengend Snippet: NMEVs effectively attenuated palmitic acid-induced senescence in C2C12 cells. (A) Schematic diagram of cell culture and treatment. (B) qRT-PCR analysis of the expression of senescence markers p53, cdkn1a, and cdkn2a in each group (n = 3). (C-C‴) Western blotting for the expression of senescence markers p53, cdkn1a (p21), and cdkn2a (p16) in each group with relative quantification (n = 6). (D-D′) Immunofluorescence staining for the DNA damage marker γH2AX with quantitative analysis (Scale bar, 100 μm; n = 6 for each group). (E-E′) Immunofluorescence staining for p16 with quantitative analysis (Scale bar, 100 μm; n = 6). (F-F′) Immunofluorescence staining for p21 with quantitative analysis (Scale bar, 100 μm; n = 6). (G-G′) Flow cytometry analysis of relative reactive oxygen species (ROS) levels (n = 6). Data are presented as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001; ns, not significant. Rel. fold, relative fold; T.Ar, total area.

Article Snippet: The C2C12 cell line was procured from Procell Life Science & Technology Co., Ltd (Wuhan, China).

Techniques: Cell Culture, Quantitative RT-PCR, Expressing, Western Blot, Quantitative Proteomics, Immunofluorescence, Staining, Marker, Flow Cytometry

Journal: Bioactive Materials

Article Title: Neonatal muscle-derived extracellular vesicles containing miR-542-3p rejuvenate aged skeletal muscle via a functional microneedle patch

doi: 10.1016/j.bioactmat.2026.06.011

Figure Lengend Snippet: NMEVs alleviated palmitic acid-induced mitochondrial dysfunction and lipid deposition. (A) Relative ATP synthesis rates in each group (n = 6). (B) qRT-PCR analysis of MT-CO1 expression in each group (n = 3). (C) qRT-PCR analysis of MT-ND1 expression in each group (n = 3). (D) qRT-PCR analysis of MT-CO3 expression in each group (n = 3). (E) qRT-PCR analysis of D-loop expression in each group (n = 3). (F) Mitochondrial complex V activity in C2C12 cells of each group (n = 6). (G) Measurement of oxygen consumption rate (OCR) in C2C12 cells of each group (n = 4). (H-H′) Transmission electron microscopy (TEM) assessment of mitochondrial quantity with quantitative analysis (Scale bar, 500 nm; n = 3). (I-I′) Western blotting for PGC-1α expression in each group with relative quantification (n = 6). (J-J′) Immunofluorescence staining for SDHA with quantitative analysis (Scale bar, 100 μm; n = 6). (K-K′) Immunofluorescence staining for EdU with quantitative analysis (Scale bar, 100 μm; n = 6). (L-L′) Representative images of BODIPY staining in each group with quantitative analysis (Scale bar, 20 μm; magnified Scale bar, 5 μm; n = 6). (M-M′) Representative images of Oil Red O staining in each group with quantitative analysis (Scale bar, 20 μm; magnified scale bar, 5 μm; n = 6). Data are presented as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: The C2C12 cell line was procured from Procell Life Science & Technology Co., Ltd (Wuhan, China).

Techniques: Quantitative RT-PCR, Expressing, Activity Assay, Transmission Assay, Electron Microscopy, Western Blot, Quantitative Proteomics, Immunofluorescence, Staining

Journal: Bioactive Materials

Article Title: Neonatal muscle-derived extracellular vesicles containing miR-542-3p rejuvenate aged skeletal muscle via a functional microneedle patch

doi: 10.1016/j.bioactmat.2026.06.011

Figure Lengend Snippet: NMEVs reduced C2C12 senescence and lipid accumulation by enriching miR-542-3p to stabilize mitochondrial function. (A) PCA plot showing sample homogeneity of AMEVs and NMEVs (n = 3). (B) Heatmap showing the top 20 significantly upregulated and downregulated microRNAs. (C) qRT-PCR analysis of the expression of the top 12 significantly upregulated microRNAs (n = 3). (D) qRT-PCR analysis of miR-542-3p expression in C2C12 cells after transfection with NMEVs, mimic, or NMEVs + inhibitor (n = 3). (E-E′) Immunofluorescence staining for p16 with quantitative analysis (Scale bar, 50 μm; n = 6). (F-F′) Immunofluorescence staining for p21 with quantitative analysis (Scale bar, 50 μm; n = 6). (G-G′) Immunofluorescence staining for γH2AX with quantitative analysis (Scale bar, 50 μm; n = 6). (H-H′) Immunofluorescence staining for EdU with quantitative analysis (Scale bar, 50 μm; n = 6). (I-I′) Immunofluorescence staining for SDHA with quantitative analysis (Scale bar, 50 μm; n = 6). (J-J‴) Western blotting for p16, p21 and PGC-1α expression in each group with relative quantification (n = 6). (K) Relative ATP synthesis rates in each group (n = 6). (L) qRT-PCR analysis of MT-CO1 expression in each group (n = 3). (M) qRT-PCR analysis of MT-ND1 expression in each group (n = 3). (N) qRT-PCR analysis of MT-CO3 expression in each group (n = 3). (O) qRT-PCR analysis of D-loop expression in each group (n = 3). (P-P′) Representative images of BODIPY staining in each group with quantitative analysis (Scale bar, 20 μm; magnified Scale bar, 10 μm; n = 6). (Q-Q′) Representative images of Oil Red O staining in each group with quantitative analysis (Scale bar, 20 μm; magnified Scale bar, 10 μm; n = 6). Data are presented as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: The C2C12 cell line was procured from Procell Life Science & Technology Co., Ltd (Wuhan, China).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Immunofluorescence, Staining, Western Blot, Quantitative Proteomics

Journal: Bioactive Materials

Article Title: Neonatal muscle-derived extracellular vesicles containing miR-542-3p rejuvenate aged skeletal muscle via a functional microneedle patch

doi: 10.1016/j.bioactmat.2026.06.011

Figure Lengend Snippet: Asxl2 and Eef1a1 served as downstream target genes of miR-542-3p. (A) Prediction of downstream target genes of miR-542-3p using multiple target gene prediction software. (B) qRT-PCR analysis of Asxl2 expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (C) qRT-PCR analysis of Eef1a1 expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (D) qRT-PCR analysis of Lrrc59 expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (E) qRT-PCR analysis of Gabarap expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (F) qRT-PCR analysis of Ap3d1 expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (G) qRT-PCR analysis of Arhgap5 expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (H) qRT-PCR analysis of Kcmf1 expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (I) qRT-PCR analysis of Pten expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (J) qRT-PCR analysis of Ube2e1 expression in C2C12 cells after transfection with miR-542-3p mimic (n = 3). (K-K″) Western blotting for Asxl2 and Eef1a1 expression in C2C12 cells after transfection with miR-542-3p mimic, with relative quantification (n = 3). (L-L′) Dual-luciferase reporter assay verifying the direct targeting binding relationship between miR-542-3p and Asxl2 (n = 3). (M-M′) Dual-luciferase reporter assay verifying the direct targeting binding relationship between miR-542-3p and Eef1a1 (n = 3). (N-N″) Western blotting for Asxl2 and Eef1a1 expression in neonatal and aging muscle tissues (n = 3). Data are presented as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: The C2C12 cell line was procured from Procell Life Science & Technology Co., Ltd (Wuhan, China).

Techniques: Software, Quantitative RT-PCR, Expressing, Transfection, Western Blot, Quantitative Proteomics, Luciferase, Reporter Assay, Binding Assay

Journal: Bioactive Materials

Article Title: Neonatal muscle-derived extracellular vesicles containing miR-542-3p rejuvenate aged skeletal muscle via a functional microneedle patch

doi: 10.1016/j.bioactmat.2026.06.011

Figure Lengend Snippet: miR-542-3p suppressed Eef1a1 to ameliorate PA-induced mitochondrial dysfunction and cellular senescence. (A-A′) Western blotting for Eef1a1 expression after PA induction, followed by transfection with miR-542-3p mimic and Eef1a1 overexpression plasmid (Eef1a1 OE ), with relative quantification (n = 3). (B) Schematic diagram illustrating Eef1a1 regulation of lipid storage via AMPK. (C-C′) Western blotting for AMPK and p-AMPK expression in neonatal and aging muscle tissues with relative quantification (n = 3). (D-D′) Western blotting for AMPK and p-AMPK expression after PA induction, followed by transfection with miR-542-3p mimic and Eef1a1 overexpression plasmid (Eef1a1 OE ), with relative quantification (n = 3). (E-E″) Representative images of p16 and p21 staining in each group with quantitative analysis (Scale bar, 50 μm; n = 6). (F) Relative ATP synthesis rates in each group (n = 6). (G) qRT-PCR analysis of MT-CO1 expression in each group (n = 3). (H) qRT-PCR analysis of MT-ND1 expression in each group (n = 3). (I) qRT-PCR analysis of MT-CO3 expression in each group (n = 3). (J) qRT-PCR analysis of D-loop expression in each group (n = 3). (K) Mitochondrial complex V activity in C2C12 cells of each group (n = 6). (L-L′) Representative images of SDHA staining in each group with quantitative analysis (Scale bar, 50 μm; n = 6). Data are presented as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: The C2C12 cell line was procured from Procell Life Science & Technology Co., Ltd (Wuhan, China).

Techniques: Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation, Quantitative Proteomics, Staining, Quantitative RT-PCR, Activity Assay

Journal: Bioactive Materials

Article Title: Bioengineered titanium implants functionalized with aptamer-valproic acid conjugates orchestrate macrophage programming and mesenchymal stem cell homing for improved osseointegration

doi: 10.1016/j.bioactmat.2026.05.055

Figure Lengend Snippet: A) Live/Dead staining of RAW 264.7 macrophages cultured on different material surfaces for 72 h, where dead cells were stained red and live cells were stained green. B) Cytoskeleton staining morphology of RAW 264.7 grown on different material surfaces for 72 h. C, D, F) Fluorescence images and flow cytometry analysis of intracellular ROS in RAW 264.7 cells with DCFH-DA probe. E) Proliferation of RAW 264.7 macrophages on different material surfaces assessed by CCK-8 assay. n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Created in BioRender.

Article Snippet: The murine macrophage cell line RAW 264.7 (Procell Life Science, China) was cultured in high-glucose DMEM (Gibco, USA) containing 10% FBS and employed for immunomodulation studies.

Techniques: Staining, Cell Culture, Fluorescence, Flow Cytometry, CCK-8 Assay

Journal: Bioactive Materials

Article Title: Bioengineered titanium implants functionalized with aptamer-valproic acid conjugates orchestrate macrophage programming and mesenchymal stem cell homing for improved osseointegration

doi: 10.1016/j.bioactmat.2026.05.055

Figure Lengend Snippet: A, F) IF staining of iNOS and CD206 in RAW 264.7 macrophages. B-E) Secretion of inflammation-related proteins in RAW 264.7 macrophages. G-J) Relative mRNA expression of inflammation-related genes in RAW 264.7 macrophages. n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The murine macrophage cell line RAW 264.7 (Procell Life Science, China) was cultured in high-glucose DMEM (Gibco, USA) containing 10% FBS and employed for immunomodulation studies.

Techniques: Staining, Expressing

Journal: Bioinorganic Chemistry and Applications

Article Title: Identifying Drug‐Induced Metabolic Signatures in Comparative Untargeted NMR Metabolomics Using Difference Spectra

doi: 10.1155/bca/9636071

Figure Lengend Snippet: Principal component analysis (PCA) of the NMR profiles. (A) PCA score plots derived from Bspectra of treated cell lysates (left, proportion of variance: PC1 (0.528), PC2 (0.276)) and respective growth media (right, proportion of variance: PC1 (0.985), PC2 (0.0068)). (B) PCA score plots of average Bdifference_spectra of cell lysates (left, proportion of variance: PC1 (0.786), PC2 (0.088)) and growth media (right, proportion of variance: PC1 (0.991), PC2 (0.0048)). (C) PCA score plot of concatenated average Bdifference_spectra (cell lysates + growth media); proportion of variance: PC1 (0.866), PC2 (0.0923). Symbols indicate cell lines: A2780 (circles) and SKOV‐3 (diamonds). Colors indicate treatment: AF (blue); AuTM (red); CN1 (green); and CN2 (yellow).

Article Snippet: The human ovarian cancer cell line SKOV‐3 (SK‐OV‐3, RRID: CVCL_0532) was obtained from CLS Cell Lines Service GmbH (Germany), Lot n. 300342‐1420SF.

Techniques: Analysis, Derivative Assay, Nuclear Magnetic Resonance

Journal: Bioinorganic Chemistry and Applications

Article Title: Identifying Drug‐Induced Metabolic Signatures in Comparative Untargeted NMR Metabolomics Using Difference Spectra

doi: 10.1155/bca/9636071

Figure Lengend Snippet: Cluster cohesion analysis of PCA derived from NMR profiles. PCA score plots derived from (A) Bspectra of treated cell lysates; (B) Bspectra of treated growth media; (C) concatenated average Bdifference_spectra (cell lysates + growth media). In each score plot, the cluster centroids are shown as black‐contoured large dots; the respective average silhouette width ( s ) and p values from the permutation tests on centroid distances ( p ) are also reported.

Article Snippet: The human ovarian cancer cell line SKOV‐3 (SK‐OV‐3, RRID: CVCL_0532) was obtained from CLS Cell Lines Service GmbH (Germany), Lot n. 300342‐1420SF.

Techniques: Analysis, Derivative Assay

Journal: Bioinorganic Chemistry and Applications

Article Title: Identifying Drug‐Induced Metabolic Signatures in Comparative Untargeted NMR Metabolomics Using Difference Spectra

doi: 10.1155/bca/9636071

Figure Lengend Snippet: Comparative analysis of metabolic dysregulation induced by the four selected gold(I) compounds and AF‐analogs. (A) Bar plots representing the dysregulation indices (average ± sd) for AF, AuTM, CN1, and CN2 in A2780 (left) and SKOV‐3 (right) cells. (C) Bar plots of the dysregulation indices (average ± sd) for all compounds in A2780 cells. Colors indicate treatments: AF (blue); AuTM (red); CN1 (green); CN2 (yellow); and AF‐analogs (AFCl, AFI, (Ph 3 P)AuI) shown as shades of blue.

Article Snippet: The human ovarian cancer cell line SKOV‐3 (SK‐OV‐3, RRID: CVCL_0532) was obtained from CLS Cell Lines Service GmbH (Germany), Lot n. 300342‐1420SF.

Techniques: Analysis

Journal: Bioinorganic Chemistry and Applications

Article Title: Identifying Drug‐Induced Metabolic Signatures in Comparative Untargeted NMR Metabolomics Using Difference Spectra

doi: 10.1155/bca/9636071

Figure Lengend Snippet: Dot plots of the pathway analysis results for the different treatments in A2780 and SKOV‐3 cells. Each enriched pathway is represented as a dot with color intensity and size reflecting the weighted average of the absolute values of the log2(FC) for the detected metabolites within each pathway, using metabolite importance as a weight. For each treatment and cell line, the pathway with the highest FC value was set to 100, and all other pathways were scaled proportionally.

Article Snippet: The human ovarian cancer cell line SKOV‐3 (SK‐OV‐3, RRID: CVCL_0532) was obtained from CLS Cell Lines Service GmbH (Germany), Lot n. 300342‐1420SF.

Techniques: Analysis

Journal: Bioinorganic Chemistry and Applications

Article Title: Identifying Drug‐Induced Metabolic Signatures in Comparative Untargeted NMR Metabolomics Using Difference Spectra

doi: 10.1155/bca/9636071

Figure Lengend Snippet: Integrated pathway and metabolite analysis of AF‐induced metabolic changes. (A) Sankey diagram showing the five most dysregulated pathways shared between A2780 and SKOV‐3 cells upon AF treatment, together with the associated metabolites. (B) Box plots of relative metabolite concentrations in the intracellular compartment (cell lysates, L). Intracellular levels of glutathione, glutamate, and glycine are shown for control (blue) and AF‐treated (cyan) A2780 and SKOV‐3 cells.

Article Snippet: The human ovarian cancer cell line SKOV‐3 (SK‐OV‐3, RRID: CVCL_0532) was obtained from CLS Cell Lines Service GmbH (Germany), Lot n. 300342‐1420SF.

Techniques: Analysis, Control

Journal: eLife

Article Title: Modulation of human dorsal root ganglion neuron excitability by Nav1.7 inhibition

doi: 10.7554/eLife.111490

Figure Lengend Snippet: Recordings were made on a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line with human Nav1.7 channels expressed in HEK293 cells using 10-hole recording wells. Sodium channel current was evoked by a step from –80 mV to +5 mV delivered every 3 s. Each concentration of AM-2099 was applied in 16 wells, and data from each well were accepted if the leak-corrected current at the holding voltage of –80 mV was less than 0.5 nA and the peak sodium current in control was >10 nA. Symbols show mean ± SD for wells meeting these criteria with current measured after 16 min of exposure to compound. ( A ) 12 wells for control solution containing 0.1% DMSO (0 nM), 8 for 1 nM, 8 for 3 nM, 8 for 10 nM, 8 for 30 nM, 8 for 100 nM, 12 for 300 nM, and 15 for 1000 nM. ( B ) 23 wells for control drug-free solution containing 0.1% DMSO, 12 for 1 nM, 13 for 3 nM, 15 for 10 nM, 9 for 30 nM, 11 for 100 nM, 11 for 300 nM, and 15 for 1000 nM. Fitted curves: Imax/(1+[Drug]/Kd), where Imax was fixed at the average value in the DMSO control wells (concentration of 0 nM) and the fit included weighting of data points by SD. Fit to data in (A) ignored the values from the column corresponding to 3 nM AM-2099 as it was obvious that there was an error in preparing this dilution. Internal solution (mM):140 CsF, 10 NaCl, 1 EGTA, 10 HEPES, 25 sucrose pH 7.3 w CsOH. External Solution (mM):145 NaCl, 4 KCl, 2 CaCl 2 , 1 MgCl 2 , 10 HEPES, 10 glucose, pH 7.4 w NaOH. All solutions contained 1 mg/mL Pluronic-F68. Figure 1—source data 1. Numerical data contained in .

Article Snippet: The concentration-dependence for inhibition of cloned human Nav1.7 channels by AM-2099 was determined using a Qube 384 automated patch-clamp instrument (Sophion Bioscience) using a stable cell line previously generated in the laboratory ( ) with human Nav1.7 channels (GenBank Accession Number X82835.1 ) expressed in a HEK293 cell line purchased from American Type Culture Collection (CAT# ATCC CRL-1573, ATCC, Manassas, VA).

Techniques: Automated, Patch Clamp, Stable Transfection, Concentration Assay, Control