Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Colorimetric staining and quantification of ALP activity in hBM-MSCs cultured for seven days on collagen type I-coated (a) and fibronectin-coated (b) substrates. Results are expressed as mean ± SD (n=3). Data were analyzed assuming a Gaussian (normal) distribution and equal variances across groups; statistical differences were assessed using one-way ANOVA. Only p-values <0.1 are shown. Scale bar represents 200 µm.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Staining, Activity Assay, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Fluorescence images of hBM-MSCs cultured on β-PVDF films coated with either collagen type I (a) or fibronectin (b) and corresponding morphological features. Nuclei are stained in blue, vinculin in green and F-actin in red. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown. Scale bars represent 200 µm.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Fluorescence, Cell Culture, Staining

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: MYPT1 phosphorylation in hBM-MSCs cultured on collagen type I-coated (a) and fibronectin-coated (b) β-PVDF films of varying surface potential. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Phospho-proteomics, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Fluorescence images of hBM-MSCs cultured on β-PVDF films coated with either collagen type I (a) or fibronectin (b) immunostained against YAP and counterstained with Hoechst 33342, and the corresponding quantifications (right panels). Scale bars represent 200 µm. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Fluorescence, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Volcano plots with the −log 10 (p-value) plotted against their respective log 2 (fold change) of genes differentially expressed in hBM-MSCs, Venn diagrams and histogram plots showing genes that are down- or upregulated (p<0.05) in hBM-MSCs cultured on the indicated surfaces for 24 hours (a) or four days (b). Circle area in a and b is proportional to the number of genes. GSEA of the cells cultured on the different surfaces for 24 hours (c) and 4 days (d).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: (a) Immunostaining of hBM-MSCs cultured on the different β-PVDF surfaces with glutaraldehyde-crosslinked collagen type I coating and corresponding morphologic analysis (b). MYTP1 phosphorylation (c) and YAP translocation (d and e) in hBM-MSCs cultured on the substrates. Only p-values <0.1 are shown. Scale bars represent 200 µm. Results are expressed as mean ± SD (n=3).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Immunostaining, Cell Culture, Phospho-proteomics, Translocation Assay

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Immunofluorescent images (a) and corresponding morphologic analysis (b) of hBM-MSCs treated with the ROCK inhibitor Y-27632 for 24 hours. YAP immunolocalization (c) and translocation (d) in treated cells. Only p-values <0.1 are shown. Scale bars represent 300 µm. Results are expressed as mean ± SD (n=4).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Translocation Assay

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Expression of neural and neurotrophic markers in nucleus pulposus cells isolated from degenerated intervertebral disc.

doi: 10.1002/jor.22098

Figure Lengend Snippet: Figure 2. Mesenchymal Profile of Human NPs-IVD: (A) Immunophenotypic profile, by citofluorimetric analysis, of nucleus pulposus from 14 human degenerated intervertebral discs. (B) NPs-IVD show mesenchymal properties under chondrogenic, adipogenic, and osteogenic differentiation.

Article Snippet: Human Mesenchymal Stem Cell Functional Identification Kit (R&D Systems, Minneapolis, MN) was used to induce adipogenic, chondrogenic, and osteogenic differentiation.

Techniques:

Journal: Molecular Therapy. Nucleic Acids

Article Title: Ratio of miRNA-29 to miRNA-199 expression coordinates mesenchymal stem cell repair of bleomycin-induced pulmonary injury

doi: 10.1016/j.omtn.2025.102461

Figure Lengend Snippet: Bleomycin-induced lung injury, which is reduced after adipose-derived mesenchymal stem cell instillation (A) Representative μCT transverse and coronal lung sections acquired from aged (22-month-old) male C57BL/6 mice at baseline (left) and 7 days following intratracheal bleomycin (BLM, 2.0 U/kg) administration (right) demonstrating increased lung density and loss of airspaces. (B) Saline treatment did not result in evidence of lung injury on μCT scan at baseline (left) or 7 days post-instillation (right). Histological sections of lung tissue collected at day 21 post-BLM were stained with Masson’s trichrome as described in . (C and D) Representative photomicrographs (20× and 40× magnifications) of lung sections from saline-treated control mice (C) and BLM-treated mice (D). (E) Infusion of adipose-derived mesenchymal stem cells (ASCs) 12 days post-BLM instillation resulted in reduced severity of pulmonary fibrosis (PF). (F) Degree of PF on histological sections was measured by semi-quantitative Ashcroft score as described in . BLM-induced lung injury resulted in increased Ashcroft score compared to saline controls. Infusion with ASCs 12 days post-BLM injury resulted in decreased Ashcroft score. (G) Intratracheal BLM instillation increased lung collagen content as measured by hydroxyproline assays as described in . Mice treated with ASCs on day 12 post-BLM had decreased lung collagen content compared to BLM-only controls. Each data point represents an individual biological replicate (mouse); n = 6–10 mice/group. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. (H) Ratio of pAKT to AKT protein expression in lung tissue of mice was quantified by western blot analysis at day 21 post-BLM sacrifice. Aged C57BL/6 mice treated with intratracheal BLM demonstrated increased pAKT/AKT protein expression compared to saline-treated controls. Lungs from mice treated with intravenous infusion of ASCs 12 days post-BLM-induced injury demonstrated decreased expression of pAKT/AKT compared to BLM-only group. Inset shows a representative western blot and β-actin loading control. Data are graphed as individual biological replicates ( n = 6–8 mice/group); ∗ p < 0.05.

Article Snippet: For mesenchymal differentiation potential, the Mouse Mesenchymal Stem Cell Functional Identification Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Derivative Assay, Saline, Staining, Control, Expressing, Western Blot

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 1. MSCs synergized with A1AT to modulate inflammation in Raw 264.7 macrophages. Cells were stimulated with 100 ng/mL LPS plus 10 ng/mL IFNγ and treated with 0.5 mg/mL A1AT or MSCs (MSC/MΦ = 1/10) or their combination. Dexamethasone (Dex, 1 µg/mL) was used as a benchmark. Pro-inflammatory mouse cytokine IL6 (A), TNFα (B), and anti-inflammatory mouse cytokine IL10 (C) were measured via ELISA. The IL6/IL10 (D) and TNFα/IL10 ratio (E) was also shown. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 2. MSCs synergized with A1AT to modulate inflammation in primary human PBMCs. Cells were stimulated with 100 ng/mL LPS + anti-CD3/CD28 antibodies (positive) and treated with 0.5 mg/mL A1AT or MSCs (MSC/PBMC = 1/10) or their combination for 24 hs. Dexamethasone (Dex, 1 µg/mL) was used as a benchmark. PBMCs without activation and treatment were used as a negative control. Pro-inflammatory human cytokine IL6 (A), TNFα (B), and anti-inflammatory human cytokine IL10 (C) were measured via ELISA. The IL6/IL10 (D) and TNFα/IL10 ratio (E) was also shown. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Activation Assay, Negative Control, Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 3. MSCs and A1AT combination treatment reduced neutrophil ROS and NETs production. HL-60 cells derived neutrophils were stimulated with 100 nM PMA and treated with 0.5 mg/mL A1AT or MSCs (MSC/neutrophil = 1/10) or their combination for 4 hs. Reactive oxygen species (ROS) (A-B) and neutrophil extracellular traps (NETs) production (C-D) were analyzed. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Derivative Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 4. MSC and A1AT combination treatment enhanced phagocytosis in THP-1 derived macrophages (A-D) and HL-60 cells derived neutrophils (E-H). Macrophages were stimulated with 100 ng/mL LPS plus 10 ng/mL IFNγ for 24 hs. Neutrophils were stimulated with 100 nM PMA for 4 hs. Cells were treated with 0.5 mg/mL A1AT or MSCs

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Derivative Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 5. MSCs synergized with A1AT to improve survival rate and reduce lung injury in mice. (A) Illustration of the model. (B) The survival rate and (C) body weight development. N = 6. (D) H&E staining and (E) lung injury scores. The lung injury scores were calculated based on the five criteria shown in (D). *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Staining

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 6. MSCs and A1AT synergized in reducing total protein (A) and pro-inflammatory cytokines while increasing anti-inflammatory cytokine IL10 (B-F) in BALF. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques:

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 8. MSCs synergized with A1AT to reduce total cell (A-B), macrophage (C), and neutrophil number (D) in BALF. The M1/M2 macrophage ratio was reduced by all treatments (E). *:p < 0.05, **:p < 0.01, ***:p < 0.001. (F-G) MSCs synergized with A1AT to reduce cell death as identified via TUNEL staining.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: TUNEL Assay, Staining

Journal: Scientific reports

Article Title: Small extracellular vesicles convey the stress-induced adaptive responses of melanoma cells.

doi: 10.1038/s41598-019-51778-6

Figure Lengend Snippet: Figure 1. Schematic illustration of the experimental workflow in six steps. B16F1 mouse melanoma cell- derived sEVs were isolated and characterised by Western Blot (WB), atomic force microscopy (AFM) and dynamic light scattering (DLS). Then B16F1 cultures were treated in five different ways, and 72 h supernatants were harvested for sEV isolation. Vesicle samples were then analysed by nanoparticle tracking analysis (NTA) to determine the number of released sEVs, sequencing and LC-MS/MS to describe their miRNome and proteome. Ingenuity Pathway Analysis (IPA) was used to analyse data and predict the functional differences between sEV groups. This in silico predictions were tested in vitro on mesenchymal stem cell (MSC) and melanoma cell cultures and MSC-B16F1 3D co-cultures as well using Ki-67-specific immunocytochemistry, Cell-Clock cell cycle assay, wound healing assay, and 3D hanging drop technology. Abbreviation: n.ctrl-negative control. Figure was created with BioRender.com.

Article Snippet: Purity of MSC cultures was checked by flow cytometry using the Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel (R&D Systems) according to the manufacturer’s instructions and a FACSCalibur instrument coupled with CellQuest Pro 6.0 (BD Biosciences).

Techniques: Derivative Assay, Isolation, Western Blot, Microscopy, Sequencing, Liquid Chromatography with Mass Spectroscopy, Functional Assay, In Silico, In Vitro, Immunocytochemistry, Cell Cycle Assay, Wound Healing Assay, Negative Control