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: Bone & Joint Research

Article Title: Naringin targets TGF-β1-mediated angiogenesis to enhance the osteogenic effect of induced membrane

doi: 10.1302/2046-3758.155.BJR-2025-0412.R1

Figure Lengend Snippet: The effect of naringin on angiogenic-osteogenic and transforming growth factor β (TGF-β)/SMAD pathway-related factors of endothelial progenitor cells (EPCs). a) The concentrations of platelet derived growth factor BB (PDGF-BB), vascular endothelial growth factor (VEGF), and slit guidance ligand 3 (SLIT3) in the supernatant of EPCs were detected by enzyme-linked immunosorbent assay. b) Alizarin red staining (ARS) was performed to detect mineralized nodules in osteoblasts. Scale bar: 50 μm. c) The protein level of p-SMAD2 and p-SMAD3 in EPCs was detected by western blot. d) was the immunofluorescence result of p-SMAD2. Scale bar: 100 μm. N = 5/group. Each value was presented as the mean (SD). ***p < 0.001 vs the control group; ##p < 0.01, ###p < 0.001 vs the small interfering RNA targeting transforming growth factor-β1 (si-TGF-β1) group.

Article Snippet: Rat osteoblasts (Procell, China) were seeded into cell culture plates.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Immunofluorescence, Control, Small Interfering RNA

Journal: Genes & Diseases

Article Title: Targeting MDK alleviates bone loss via dual regulation of osteogenic differentiation and inflammatory cytokine expression

doi: 10.1016/j.gendis.2025.101931

Figure Lengend Snippet: Recombinant MDK protein inhibits osteogenic differentiation in vitro in a dose-dependent manner. (A) Cell viability after treating MC3T3-E1 cells with recombinant MDK protein after 48 h, assessed using the CCK-8 assay. Inter-group comparisons were analyzed by one-way ANOVA. (B) Western blotting analysis of ALP, RUNX2, OSX, and OCN expression levels following MDK treatment (7 days). (C–F) Reverse transcription PCR analysis of mRNA expression levels of Alpl , Runx2 , Sp7 , and Bglap in MC3T3-E1 cells following MDK treatment (7 days). β-actin served as the internal control. Inter-group comparisons were analyzed by one-way ANOVA. (G, H) ALP staining and activity assays were performed after inducing MC3T3-E1 cells with recombinant MDK protein (0–600 ng/mL) for 14 days. Inter-group comparisons were analyzed by one-way ANOVA. (I, J) ARS staining and quantitative analysis were conducted after inducing MC3T3-E1 cells with recombinant MDK protein (0–600 ng/mL) for 21 days. Inter-group comparisons were analyzed by one-way ANOVA. Scale bar, 100 μm ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; “ns” indicates non-significant differences.

Article Snippet: Procell Life Science & Technology Co., Ltd. (Wuhan, China) supplied the pre-osteoblastic cell line MC3T3-E1 Subclone 14.

Techniques: Recombinant, In Vitro, CCK-8 Assay, Western Blot, Expressing, Reverse Transcription, Control, Staining, Activity Assay

Journal: Genes & Diseases

Article Title: Targeting MDK alleviates bone loss via dual regulation of osteogenic differentiation and inflammatory cytokine expression

doi: 10.1016/j.gendis.2025.101931

Figure Lengend Snippet: MDK suppresses osteoblast differentiation via the PI3K/AKT signaling pathway. (A, B) Western blot detection of the effect of recombinant MDK protein on the protein expression of molecules in the PI3K/AKT signaling pathway during the differentiation of MC3T3-E1 to osteoblasts (7 days). Inter-group comparisons were analyzed by a two-tailed unpaired Student's t -test (for normally distributed data with equal variance). (C, D) ALP, RUNX2, and OSX expression levels were detected by Western blotting. MC3T3-E1 cells were pretreated with 30 μM LY294002. Osteogenic differentiation was induced for 7 days. Inter-group comparisons were analyzed by one-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; “ns” indicates non-significant differences.

Article Snippet: Procell Life Science & Technology Co., Ltd. (Wuhan, China) supplied the pre-osteoblastic cell line MC3T3-E1 Subclone 14.

Techniques: Western Blot, Recombinant, Expressing, Two Tailed Test

Journal: Genes & Diseases

Article Title: Targeting MDK alleviates bone loss via dual regulation of osteogenic differentiation and inflammatory cytokine expression

doi: 10.1016/j.gendis.2025.101931

Figure Lengend Snippet: Recombinant MDK protein triggers the activation of inflammatory cytokines through the NF-κB signaling pathway. (A, B) IL-6, TNFα, and IL-1β expression levels were detected using Western blotting. MC3T3-E1 cells were treated with recombinant MDK protein (600 ng/mL). Osteogenic differentiation was induced for 7 days. Inter-group comparisons were analyzed by a two-tailed unpaired Student's t -test (for normally distributed data with equal variance). (C, D) Western blotting analysis of NF-κB signaling pathway molecules in MC3T3-E1 cells treated with recombinant MDK protein for 7 days during osteoblastic differentiation. Inter-group comparisons were analyzed by two-tailed unpaired Student's t -test (for normally distributed data with equal variance). (E, F) IL-6 and IL-1β expression levels were detected using Western blotting. MC3T3-E1 cells were pretreated with 10 μM BAY 11–7082. Osteogenic differentiation was induced for 7 days. Inter-group comparisons were analyzed by one-way ANOVA. ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: Procell Life Science & Technology Co., Ltd. (Wuhan, China) supplied the pre-osteoblastic cell line MC3T3-E1 Subclone 14.

Techniques: Recombinant, Activation Assay, Expressing, Western Blot, Two Tailed Test

Journal: ACS Omega

Article Title: Biphasic Bone Implants through Hybrid Extrusion Printing of Thermoplastic Poly(lactic- co -glycolic) acid and Strontium-Modified Calcium Phosphate Bone Cement

doi: 10.1021/acsomega.5c12496

Figure Lengend Snippet: Monophasic SrCPC and PLLA–PGA scaffolds and biphasic PLLA–PGA/SrCPC scaffolds seeded with SAOS-2. Printed scaffolds used for cell culture experiments; scale bars = 2 mm (a). Overview fluorescence microscopic images of scaffolds seeded with 5 × 10 5 cells/scaffold after 1 and 14 days of cultivation (b); adherent cells were stained with DAPI (cell nuclei; blue) and phalloidin (actin cytoskeletons; green), scale bars = 500 μm, C = cement (SrCPC), P = polymer (PLLA–PGA). Cell attachment and density on SrCPC and PLLA–PGA strands in monophasic and biphasic scaffolds seeded with 5 × 10 5 cells/scaffold (c) or 5 × 10 4 cells/scaffold (d). Detailed fluorescence microscopic images of scaffolds after 1 and 14 days of cultivation. Cells were stained with DAPI (cell nuclei; blue) and phalloidin (actin cytoskeletons; green). Scale bars = 100 μm.

Article Snippet: The osteoblast-like cell line SaOS-2 (ATCC 243; DSMZ, Braunschweig, Germany) was used to study the cell response to biphasic scaffolds in a direct contact culture.

Techniques: Cell Culture, Fluorescence, Staining, Polymer, Cell Attachment Assay

Journal: ACS Omega

Article Title: Biphasic Bone Implants through Hybrid Extrusion Printing of Thermoplastic Poly(lactic- co -glycolic) acid and Strontium-Modified Calcium Phosphate Bone Cement

doi: 10.1021/acsomega.5c12496

Figure Lengend Snippet: Cell growth and ALP activity of SAOS-2 cells cultured on monophasic SrCPC and PLLA–PGA scaffolds in comparison with biphasic PLLA–PGA/SrCPC scaffolds. Cell number was correlated with the cytosolic LDH activity measured after cell lysis (a), and the ALP activity as an indicator of osteogenic differentiation is shown as absolute activity per scaffold (b) and as specific activity in relation to the cell number (c) ( n = 3, mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The osteoblast-like cell line SaOS-2 (ATCC 243; DSMZ, Braunschweig, Germany) was used to study the cell response to biphasic scaffolds in a direct contact culture.

Techniques: Activity Assay, Cell Culture, Comparison, Lysis