murine pre osteoblasts  (ATCC)

Journal: Bioactive Materials

Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

doi: 10.1016/j.bioactmat.2026.02.005

Figure Lengend Snippet: Curvature-controlled orientation of cytoskeletal stress fibers on concave-cylindrical surfaces. (a) Representative confocal microscopy images depicting F-actin (magenta) and nuclei (blue) of fibroblasts, mesenchymal stromal cells, osteoblasts, pre-osteoblasts and endothelial cells seeded on flat surfaces. (b) Brass mold used to fabricate the master GeoChip from which GeoChips for use in cell culture are manufactured via sugar candy molding . Photographs show the topographic surface of the brass mold and the candy mold (Scale bar: 2 mm). Scanning electron microscopy (SEM) verified the smoothness of the resulting curved surface (half-cylinder with Ø = 1000 μm, scale bar: 200 μm). (c) Representative confocal microscopy images of cells seeded on concave-cylindrical surfaces with Ø = 100 and 1000 μm. Yellow dashed lines indicate the half-cylinder boundaries. (d-i) Distribution of stress fiber orientation quantified from the F-actin signal of cells on substrates with increasing curvature (average with standard deviation). Cartesian plots include data for fibroblasts (blue), mesenchymal stromal cells (green), osteoblasts (purple), pre-osteoblasts (orange) and endothelial cells (red). The direction 0° - 180° represents the orientation along the cylindrical surface (minimum curvature) and the direction 90° represents the orientation perpendicular to the cylindrical surface (maximum curvature). The substrate curvature experienced in dependency of the orientation is indicated by the red dashed line and red scale. Random orientation is indicated by the black dashed line. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/cell type, 1 donor/cell type. Scale bars 100 μm (unless otherwise stated).

Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

Techniques: Confocal Microscopy, Cell Culture, Electron Microscopy, Standard Deviation, MANN-WHITNEY

Journal: Bioactive Materials

Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

doi: 10.1016/j.bioactmat.2026.02.005

Figure Lengend Snippet: Incidence of cell spanning on concave-cylindrical surfaces. (a) Lateral view of fibroblasts exposed to cylinders with increasing diameter (decreasing curvature), with spanning cells marked by yellow arrows. (b) Probability of spanning cells in relation to the half-cylinder diameter. (c-e, top to bottom) Representative 3D reconstructed images of fibroblasts, pre-osteoblasts and endothelial cells on concave-cylindrical surfaces with Ø = 100, 200 and 300 μm. Cells were reconstructed in Imaris using the F-actin (magenta, cell surface reconstruction) and nuclei (blue) signal as obtained by confocal microscopy. Half-cylinder contour is indicated by the yellow dashed line. Spanning cells are indicated by yellow arrows in subfigures c-e for clarity. Polar plots on the right depict the percentage of spanning cells and the corresponding angle of cell orientation for fibroblasts (blue), pre-osteoblasts (orange) and endothelial cells (red). The direction 0° - 180° represents the orientation along the cylindrical surface (minimum curvature) and the direction −90° - 90° represents the orientation perpendicular to the cylindrical surface (maximum curvature). (f) Confocal microscopy images of representative cell morphologies for fibroblasts, pre-osteoblasts and endothelial cells depicting F-actin (magenta), nuclei (blue) and focal adhesions via vinculin staining (green). Focal adhesions are indicated by green arrows (example shown on fibroblasts). (g) Cell length quantified as the major axis of an ellipse fitted around the cell. (h) Cell roundness with a value of 1 representing a perfect circle and value of 0 representing a straight line. (i) FSD calculated as the distance between FA clusters (see methods part for detailed description). (j) FA size distribution per cell plotted as the percentage of FAs that fall into the indicated size classes. (k) Representative force vector maps and (l) total cell force quantified via TFM. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/cell type. N ≥ 60 cells/cell type for FA and morphological analysis. 1 donor/cell type. Scale bar 50 μm.

Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

Techniques: Confocal Microscopy, Staining, Plasmid Preparation, MANN-WHITNEY

Journal: Bioactive Materials

Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

doi: 10.1016/j.bioactmat.2026.02.005

Figure Lengend Snippet: Cell spanning initiates channel closure and subsequent tissue remodeling. (a) Fabrication of full-cylindrical channels with Ø = 250 μm in PDMS substrates by direct molding from a micro-machined brass mold. (b) Degree of channel closure representing the distribution of cells within the channels at the selected points in time during live confocal imaging. A value of 0 indicates that cells are exclusively found at the wall of the channel and a value of 1 indicates cells have completely closed the channel and are homogeneously distributed. (c) Relative degree of alignment of the cell-network within the channels quantified as the maximum value of the orientation distribution for the individual cell types and time points normalized to the highest detected value of all conditions (see also Supplementary Data S2). Higher values indicate a higher degree of alignment along the channel axis. (d-f) Lateral and front view of the PDMS cylindrical channels obtained by live confocal imaging of fibroblasts (blue), pre-osteoblasts (orange) and endothelial cells (red) using CellTracker™ Green ( t = 4, 12, 24 and 48 h after seeding). Open arrows indicate cells spanning perpendicular to the channel axis. Full arrows indicate cells oriented along the direction of the channel axis after channel closure. Channel contour is highlighted by the yellow dashed lines. The surface of the forming tissue is marked by red dashed lines. White dashed lines indicate the z-volume that is shown in the corresponding lateral views. Statistical significance via Mann-Whitney test with Bonferroni correction, ∗p < 0.05. N = 3 cylindrical channels/cell type. 1 donor/cell type. Scale bars 100 μm.

Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

Techniques: Imaging, MANN-WHITNEY

Journal: Bioactive Materials

Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

doi: 10.1016/j.bioactmat.2026.02.005

Figure Lengend Snippet: Channel closure mechanism can be controlled by substrate curvature using scaffolds with well-defined geometries. (a, left) Schematic representation of the in vitro culture setup with collagen scaffold presenting channels of controlled diameter with Ø ≈ 600 μm, Ø ≈ 350 μm and Ø ≈ 150 μm. Monolayer seeding on one side of the biomaterial facilitates migration of cells from one end of the biomaterial. (a, right) SEM image of the microarchitecture (Scale bar 20 μm) and channels within the biomaterial (Scale bars 100 μm). SEM images correspond to the outermost surface of the scaffold. (b) Comparison of template diameter against resulting channel diameter after cross-linking and sterilization of the biomaterial. (c) Representative images of fibroblasts, pre-osteoblasts and endothelial cells within channels of distinct diameters 7 days after seeding. Cell cytoskeleton (F-actin) is depicted in magenta and nuclei in blue. Yellow arrows indicate the direction (arrow angle) and degree of alignment (vector length) for the corresponding region. Scale bar close-up images: 25 μm. (d, left) Degree of channel closure for the investigated channel diameters and cell types. (d, right) Relative degree of tissue alignment for the different channel diameters and cell types. Tissue alignment ranges from 0 (fully isotropic) to 1 (fully anisotropic, dashed line). Tissue across the channel and relative degree of is calculated in the central 50 % of each channel. Data displayed as average with standard deviation. N = 4 scaffolds/cell type. 1 donor/cell type. Scale bars 200 μm (unless otherwise stated).

Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

Techniques: In Vitro, Migration, Comparison, Plasmid Preparation, Standard Deviation

Journal: Scientific Reports

Article Title: Understanding of the different roles of Noggin in the Noggin-BMP-2 and Noggin-BMP-9 dimer complexes at the molecular level

doi: 10.1038/s41598-025-33735-8

Figure Lengend Snippet: Initial and final structures of Noggin-BMP-2 and Noggin-BMP-9 after 100 ns MD simulation. Identification of the binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 Cells by Noggin using Immunoprecipitation. ( A ) Initial structures of Noggin-BMP-2 and ( B ) Initial structures of Noggin-BMP-9 ( C ) Aligned structures of initial (White color) and final structures of Noggin-BMP-2 (Coral/Blue) ( D )Aligned structures of initial (White) and final structures of Noggin-BMP-9 (Green/Ultraviolet) ( E ) RMSD of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). ( F ) RMSF (Root Mean Squared Fluctuation) of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). RMSD and RMSF values ​​represent the mean and standard deviation of three independent simulations. ( G ) Western blot analysis using total cell lysates. ( H ) Immunoprecipitation (IP) of BMPR2 followed by immunoblotting (IB) to detect BMP-2 and BMP-9 binding.

Article Snippet: The mouse pre-osteoblastic cell line MC3T3-E1 was purchased from the American Type Culture Collection (ATCC, CRL-2593, Manassas, VA, USA).

Techniques: Binding Assay, Immunoprecipitation, Standard Deviation, Western Blot

Journal: Scientific Reports

Article Title: Understanding of the different roles of Noggin in the Noggin-BMP-2 and Noggin-BMP-9 dimer complexes at the molecular level

doi: 10.1038/s41598-025-33735-8

Figure Lengend Snippet: Identification of binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 cells by Noggin. ( A ) BMP-2–treated group, ( B ) BMP-2 + Noggin–treated group, ( C ) BMP-9–treated group, ( D ) BMP-9 + Noggin–treated group. DAPI (blue), anti-BMPR2 (red), and anti-BMP-2 or anti-BMP-9 (green). Merged images show co-localization (yellow). Scale bar: 10 μm; magnification: ×600. ( E ) Western blot of SMAD1/5/9 phosphorylation under the indicated treatments. Top, p-SMAD1/5/9; middle, total SMAD1/5/9; bottom, GAPDH. ( F) Alkaline phosphatase (ALP) activity measured 60 min after BMP addition using a p-nitrophenyl phosphate colorimetric assay (405 nm). Bars indicate mean ± SD ( n ≥ 3). p < 0.0001 vs. NT for all treated groups; asterisks denote significance relative to NT.

Article Snippet: The mouse pre-osteoblastic cell line MC3T3-E1 was purchased from the American Type Culture Collection (ATCC, CRL-2593, Manassas, VA, USA).

Techniques: Binding Assay, Western Blot, Phospho-proteomics, Activity Assay, Colorimetric Assay