human primary bone marrow cd34 cells  (ATCC)

Journal: Polymers

Article Title: Osteoinductive and Biocompatibility Assessment of a 3D-Printed Polymeric–Hydroxyapatite Composite Interference Screw

doi: 10.3390/polym18101239

Figure Lengend Snippet: Scanning electron microscopy characterization of mesenchymal stem cell attachment and proliferation on three biomaterial compositions. Top panels ( A – C ): Biomaterial surfaces prior to cell seeding, showing the characteristic surface morphology of Group 1 (PCL + HA, 60/40) ( A ), Group 2 (PCL + PLGA, 50/50) ( B ), and Group 3 (PCL + PLGA + HA, 30/30/40) ( C ) respectively. Bottom panels ( D – F ): Mesenchymal stem cells after 21 days of culture in osteogenic differentiation medium, demonstrating robust cell attachment, spreading, and surface colonization on Groups 1 ( D ), 2 ( E ), and 3 ( F ), respectively. Panel ( G ): Higher magnification view (×3500) illustrating characteristic elongated morphology of healthy, proliferative mesenchymal stem cells with extensive filopodia and lamellipodia formation extending across the Group 3 biomaterial surface, indicating strong cell–substrate interactions and active synthetic activity.

Article Snippet: Human bone marrow-derived mesenchymal stem cells were obtained from PromoCell (Heidelberg, Germany) and cultured in Mesenchymal Stem Cell Growth Medium MSC2 according to the manufacturer’s instructions.

Techniques: Electron Microscopy, Cell Attachment Assay, Activity Assay

Journal: Polymers

Article Title: Osteoinductive and Biocompatibility Assessment of a 3D-Printed Polymeric–Hydroxyapatite Composite Interference Screw

doi: 10.3390/polym18101239

Figure Lengend Snippet: Alamar Blue assay evaluating mesenchymal stem cell metabolic activity on three 3D-printed biomaterial compositions over time. ( A ) Line graph showing percentage of cell metabolic activity measured at days 3, 7, 14, and 21 on PCL + HA (60/40), PCL + PLGA (50/50), and PCL + PLGA + HA (30/30/40) substrates. Days 3, 7, 14, and 21 refer to the duration of osteogenic differentiation culture, defined as Day 0 of osteogenic induction initiated after 3 days of proliferation medium pre-culture. ( B ) Corresponding bar graph representation of metabolic activity for each composition at individual time points (D3, D7, D14, D21). Data are presented as mean ± SEM (n = 3 biological replicates per group per timepoint). Statistical annotations indicate comparisons between material groups as follows: * comparison between PCL + PLGA + HA and PCL + HA; # comparison between PCL + PLGA + HA and PCL + PLGA in A. Normality of residuals was assessed using the Shapiro–Wilk test, and homogeneity of variances was assessed using Levene’s test; assumptions were met for all groups (Shapiro p > 0.05; Levene p > 0.05). Statistical differences were evaluated by two-way ANOVA, followed by Tukey multiple comparisons with Bonferroni correction.#: PCL + PLGA + HA vs. PCL + PLGA (# p < 0.05, ## p < 0.01, (n = 3 per group)) *: PCL + PLGA + HA vs. PCL + HA ( p < 0.05 (*), p < 0.01 (**), p < 0.001 (***)).

Article Snippet: Human bone marrow-derived mesenchymal stem cells were obtained from PromoCell (Heidelberg, Germany) and cultured in Mesenchymal Stem Cell Growth Medium MSC2 according to the manufacturer’s instructions.

Techniques: Alamar Blue Assay, Activity Assay, Comparison

Journal: Polymers

Article Title: Osteoinductive and Biocompatibility Assessment of a 3D-Printed Polymeric–Hydroxyapatite Composite Interference Screw

doi: 10.3390/polym18101239

Figure Lengend Snippet: Relative gene expression of osteogenic markers in MSCs cultured on Implant A and Implant B 3D-printed PCL + PLGA + HA (30/30/40) composite scaffolds at days 14 and 21 of osteogenic differentiation. Days 14 and 21 refer to the duration of osteogenic differentiation culture, initiated after 3 days of proliferation medium pre-culture. Implant A: surface-coated PCL + PLGA + HA (30/30/40); Implant B: uncoated PCL + PLGA + HA (30/30/40) control. ( A ) Alkaline phosphatase (ALP), ( B ) Runt-related transcription factor 2 (RUNX2), and ( C ) Bone gamma-carboxyglutamate protein (BGLAP). Gene expression was normalized to GAPDH using the 2 −ΔΔCt method. Data are presented as mean ± SD (n = 4 biological replicates, each analyzed with three technical replicates). Normality of residuals was assessed using the Shapiro–Wilk test, and homogeneity of variances was assessed using Levene’s test; assumptions were met for all genes (Shapiro p > 0.05; Levene p > 0.05). Statistical differences were evaluated gene-by-gene using a two-way ANOVA (Time_Point: Day 14 vs. Day 21; Condition: Implant A vs. Implant B) including the interaction term (Time_Point × Condition), followed by Tukey/emmeans post hoc multiple comparisons with Bonferroni correction. Significance is indicated as follows: p < 0.05 (*) and p < 0.001 (***).

Article Snippet: Human bone marrow-derived mesenchymal stem cells were obtained from PromoCell (Heidelberg, Germany) and cultured in Mesenchymal Stem Cell Growth Medium MSC2 according to the manufacturer’s instructions.

Techniques: Gene Expression, Cell Culture, 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 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: 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