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: Biology

Article Title: Grape Pomace Polyphenolic Extract Promotes Osteogenic Differentiation in Human Mesenchymal Stem Cells Through Activation of RUNX2 and NRF2 Transcription Factors: A Potential Natural Strategy for Osteoporosis Prevention

doi: 10.3390/biology15090719

Figure Lengend Snippet: Effects of GPE on osteoblast differentiation of human MSCs. BMSCs and AdMSCs were cultured in basal medium (CTR) or osteogenic differentiation medium (OM) with or without GPE (1–10 µg GAE/mL) for up to 21 days, and matrix mineralization was analyzed by Alizarin Red S staining. Representative light microscopic images of Alizarin Red S staining in BMSCs ( A ) and AdMSCs ( B ) and quantitative analysis ( C ). Each experiment was performed in triplicate. All data are presented as mean ± standard deviation (SD). # p < 0.01 versus undifferentiated control cells (CTR); * p < 0.05 versus osteogenic differentiated cells (OM).

Article Snippet: Human bone marrow-derived mesenchymal stem cells (BMSCs) from healthy donors (ATCC-PCS-500-012) were purchased from ATCC (Milan, Italy) as well as mesenchymal stem cell basal medium (ATCC PCS500030) and mesenchymal stem cell growth kit for BMSC (ATCC PCS500041).

Techniques: Cell Culture, Staining, Standard Deviation, Control

Journal: Biology

Article Title: Grape Pomace Polyphenolic Extract Promotes Osteogenic Differentiation in Human Mesenchymal Stem Cells Through Activation of RUNX2 and NRF2 Transcription Factors: A Potential Natural Strategy for Osteoporosis Prevention

doi: 10.3390/biology15090719

Figure Lengend Snippet: Effects of GPE on the adipogenic differentiation of human MSCs. BMSCs and AdMSCs were cultured in basal medium (CTR) or adipogenic differentiation medium (AM) with or without GPE (1–10 µg GAE/mL) for up to 21 days, and lipid droplet formation was analyzed by Oil Red O (ORO) staining. Representative light microscopic images of ORO staining in BMSCs ( A ) and AdMSCs ( B ) and quantitative analysis ( C ). Each experiment was performed in triplicate. All data are presented as mean ± standard deviation (SD). # p < 0.01 versus undifferentiated control cells (CTR); * p < 0.05 and ** p < 0.01 versus adipogenic differentiated cells (AM).

Article Snippet: Human bone marrow-derived mesenchymal stem cells (BMSCs) from healthy donors (ATCC-PCS-500-012) were purchased from ATCC (Milan, Italy) as well as mesenchymal stem cell basal medium (ATCC PCS500030) and mesenchymal stem cell growth kit for BMSC (ATCC PCS500041).

Techniques: Cell Culture, Staining, Standard Deviation, Control

Journal: bioRxiv

Article Title: TAR syndrome causal gene RBM8A is critical for embryonic bone development and proper Hedgehog signaling

doi: 10.64898/2026.04.21.718480

Figure Lengend Snippet: A. Immunofluorescence staining of cultured human bone marrow-derived mesenchymal stem cells showing CD90 (green) and RBM8A (red). Merged image (right) demonstrates RBM8A localization in CD90-positive cells. Scale bar = 50 µm. B. Whole-mount immunofluorescence staining of E10.5 mouse embryo section showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is broadly expressed, with high expression in the limb bud. In the neural crest, RBM8A expression is decreased in SOX9-positive regions. Scale bar = 200 µm. C. Whole-mount immunofluorescence staining of E14.5 mouse forelimb autopod sections showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is broadly distributed throughout the autopod, including SOX9-positive cartilage condensation. Scale bar = 200 µm. D. Whole-mount immunofluorescence staining of E14.5 mouse forelimb zeugopod longitudinal tissue section showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is detected in and around SOX9-positive skeletal condensations, including the central region of the developing cartilage element. Scale bar = 200 µm.

Article Snippet: Human bone marrow-derived MSCs were obtained from ATCC (catalog #: PCS-500-012) and cultured using Bone Marrow-Mesenchymal Stem Cell Basal Medium (ATCC, catalog #: PCS-500-030) with 7% FBS and growth factor supplements (15 ng/ml IGF-1, 125 ng/ml FGF2).

Techniques: Immunofluorescence, Staining, Cell Culture, Derivative Assay, Expressing