Journal: bioRxiv

Article Title: BMP signaling orchestrates a transcriptional network to control the fate of mesenchymal stem cells (MSCs)

doi: 10.1101/104927

Figure Lengend Snippet: (A-D) LacZ staining (blue) of sagittal sections of mandibular molars from Gli1-LacZ mice at P0.5, P3.5, P7.5, and P21.5. Arrowheads indicate Gli1+ cells in the apical region of the mesenchyme. (E-H) Visualization of sagittal sections of mandibular molars from Gli1-CreER;tdTomato mice at P4.5 and P18.5 after induction at P3.5. The progeny of the Gli1 lineageappear red. Boxes in E and G are shown magnified in F and H, respectively. (I-L) Colony forming assay and osteogenic, chondrogenic, and adipogenic differentiation assays of cells from the Gli1+ region in the apical mesenchyme of molars from P5.5 Gli1-CreER;tdTomato mice induced at P3.5. Toluidine blue staining was used to visualize colony formation after culture for two weeks (I). Insert shows colonies are derived from Gli1+ cells (red). Alizarin Red (J), Alcian Blue (K), and Oil Red O (L) staining to detect osteogenic, chondrogenic, and adipogenic differentiation after three weeks. Scale bars, 100μm.

Article Snippet: For the differentiation assays, cells from the colonies were cultured until confluent and then induced in osteogenic, adipogenic or chondrogenic differentiation medium (05504, 05503, 05455, Stemcell Technologies, Vancouver, Canada) for 2-3 weeks according to the manufacturer’s instructions.

Techniques: Staining, Derivative Assay

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: Osteogenic differentiation promoted by chemical and mechanical stimulation. (A) Western blotting analysis shows that the expression levels of runt‐related transcription factor 2 (RUNX2) and osteopontin (OPN) increase in hASCs following chemical‐induced osteogenic differentiation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. (B) ALP staining assay results reveal a progressive increase in ALP activity during the osteogenic differentiation of hASCs. Scale bar = 200 μm. (C) Immunofluorescence staining confirms an increase in RUNX2 expression during the osteogenic differentiation of hASCs. Scale bar = 20 μm. (D) Western blotting analysis shows that the expression levels of RUNX2 and OPN increased in hASCs following mechanical stimulation. (E) ALP staining assay results reveal a progressive increase in ALP activity under mechanical stimulation. Scale bar = 200 μm. (F) Immunofluorescence staining confirms an increase in RUNX2 expression during the mechanical stimulation of hASCs. Scale bar = 20 μm.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: Western Blot, Expressing, Staining, Activity Assay, Immunofluorescence

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: Effect of PDLIM5 on osteogenic differentiation of stem cells. (A) The volcano plot shows the gene information between the positive and negative osteogenic‐related genes. (B) The heat map shows the 998 differentially expressed genes. The pink colour indicates upregulated genes, and the blue indicates downregulated genes. Group GM: Cells treated with growth medium. Group OS: Cells treated with osteogenic differentiation medium. p < 0.05, logFC > 1.2. (C) Western blotting analysis shows the expression levels of PDLIM5, α‐actinin 1 and lamin A in hASCs following chemical‐induced osteogenic differentiation. (D, E) Immunofluorescence staining confirms an increase in the expression of PDLIM5, α‐actinin1 and lamin A during osteogenic differentiation of hASCs. Scale bar = 20 μm. (F) Western blotting analysis shows the expression levels of PDLIM5, α‐actinin 1 and lamin A protein increased in hASCs following mechanical stimulation. (G–I) Immunofluorescence staining confirms an increase in the expression of PDLIM5, α‐actinin1 and lamin A during the mechanical stimulation of hASCs. Scale bar = 50 μm.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: Downregulation of PDLIM5 impairs osteogenic differentiation induced by chemical and mechanical stimulation. (A) Transfection efficiency of the negative control transfected (NC1 group) and PDLIM5 knockdown transfected (shPDLIM5 group) in hASCs. The transfection efficiency was evaluated based on the GFP fluorescence intensity. (B) ALP staining intensity is significantly reduced in shPDLIM5 group compared with the NC1 group when subjected to osteogenic differentiation. (C) Immunofluorescence staining confirms significantly reduced PDLIM5 expression levels in shPDLIM5 group compared with NC1 group. Scale bar = 20 μm. (D) Western blot analysis demonstrates the expression levels of OPN, RUNX2, PDLIM5, α‐actinin1 and lamin A in the NC1 group and shPDLIM5 group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. (E–G) Immunofluorescence staining confirms that lentivirus‐mediated PDLIM5 knockdown decreased the expression levels of RUNX2, α‐actinin1 and lamin A in hASCs following chemical‐induced osteogenic differentiation. Scale bar = 20 μm. (H) Western blotting analysis demonstrates the expression levels of OPN, RUNX2, PDLIM5, α‐actinin1 and lamin A in NC1 group and shPDLIM5 group under cyclic stretch loading.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: Transfection, Negative Control, Knockdown, Fluorescence, Staining, Immunofluorescence, Expressing, Western Blot

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: Bioinformatics analysis of PDLIM5 upstream protein and differential gene expression in osteogenic differentiation. (A) Protein–Protein Interaction network of serpin E2 interacting with integrin β3‐PDLIM5‐lamin A. (B) Western blot analysis shows the expression levels of serpin E2 and integrin β3 in NC1 group and shPDLIM5 group induced by chemical. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. (C) Western blotting analysis shows the expression levels of serpin E2 and integrin β3 in NC1 group and shPDLIM5 group under cyclic stretch loading. (D) UMAP clustering results show the top 30 principal components and the distribution of hASCs‐GM and hASCs‐OS samples. Uniform manifold approximation and projection (UMAP) plots show the origin of the hASCs‐GM cells ( n = 3687) and the hASCs‐OS cells ( n = 5511) and the 10 cell clusters (0–9). (E) The distribution of subpopulations of cells in the two samples (hASCs‐GM and hASCs‐OS) based on the integrated data from the UMAP clustering results. (F) UMAP clustering of the subgroup of cells based on the integrated data from the two samples. (G) Distribution of the number of cells in each subgroup based on the integrated data from the UMAP clustering results. (H) Heatmap shows the top five tag genes in different subgroups. Each row represents a gene; each column represents a single cell; colour depth represents the intensity of gene expression in the samples from the hASCs‐GM and hASCs‐OS groups.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: Gene Expression, Western Blot, Expressing

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: Effects of serpin E2 on PDLIM5 and integrin β3 during osteogenic differentiation. (A) Western blotting results show the expression levels of serpin E2 and integrin β3 in the hASCs following chemical‐induced osteogenic differentiation. (B, C) Immunofluorescence staining confirms an increase in serpin E2 and integrin β3 expression during osteogenic differentiation of hASCs. Scale bar = 20 μm. (D) Western blotting results show that the expression levels of serpin E2 and integrin β3 increased in hASCs following mechanical stimulation. (E, F) Immunofluorescence staining confirms an increase in the expression of serpin E2 and integrin β3 during the mechanical stimulation of hASCs. Scale bar = 50 μm. (G) Transfection efficiency of the negative control transfected (NC2 group) and serpin E2‐overexpressing transfected (OE‐Serpin E2 group) in hASCs. The transfection efficiency was evaluated based on the GFP fluorescence intensity. (H) Serpin E2‐overexpressing hASCs exhibit significantly higher ALP activity compared with the NC2 group when subjected to osteogenic differentiation. (I) Immunofluorescence analysis reveals that serpin E2 expression levels were significantly higher in the OE‐Serpin E2 group compared with the NC2 group following osteogenic differentiation. Scale bar = 20 μm. J. Western blotting results show the expression levels of serpin E2, integrin β3 and PDLIM5 in NC2 group and OE‐Serpin E2 group when subjected to osteogenic differentiation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. (K) Western blotting analysis shows the expression levels of serpin E2, integrin β3 and PDLIM5 in NC2 group and the OE‐Serpin E2 group under cyclic stretch loading. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Transfection, Negative Control, Fluorescence, Activity Assay

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: Overexpression of Serpin E2 affects the regulation of the signalling axis on the osteogenic differentiation of stem cells. (A) Western blotting analysis demonstrates the expression levels of OPN, RUNX2, serpin E2 and integrin β3 in NC2 group and OE‐Serpin E2 group during osteogenic differentiation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. (B, C) Immunofluorescence analysis confirms that lentivirus‐mediated overexpression of serpin E2 enhances the expression of RUNX2 and integrin β3 in hASCs during osteogenic differentiation. (D) Western blotting analysis reveals the expression levels of OPN, RUNX2, serpin E2 and integrin β3 in the NC2 group and OE‐Serpin E2 group under cyclic stretch loading. (E) Western blotting analysis demonstrates the expression levels of α‐actinin1 and lamin A in the NC2 group and OE‐Serpin E2 group during osteogenic differentiation. (F) Western blotting analysis demonstrates the expression levels of α‐actinin1 and lamin A in the NC2 group and OE‐Serpin E2 group under cyclic stretch loading. (G, H) Immunofluorescence analysis confirms that lentivirus‐mediated overexpression of serpin E2 enhances the expression of α‐actinin1 and lamin A in hASCs during osteogenic differentiation.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: Over Expression, Western Blot, Expressing, Immunofluorescence

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: Effect of integrin β3 deletion on SerpinE2/PDLIM5 axis‐mediated bone regeneration in vivo. (A) Western blotting results show that the expression levels of OPN, RUNX2, integrin β3, serpin E2, PDLIM5, α‐actinin1 and lamin A in mouse adipose‐derived stem cells (mASCs) following chemical‐induced osteogenic differentiation. B. Western blot analysis reveals the expression levels of OPN, RUNX2, integrin β3, serpin E2, PDLIM5, α‐actinin1 and lamin A in mASCs isolated from wild‐type (WT) and integrin β3 knockout (KO) mice. (C) H&E staining results indicate that the thickness and continuity of the trabecular bone are significantly reduced in integrin β3 KO mice compared to WT mice. Scale bar = 50 μm. (D) Representative 3D reconstruction, micro‐CT analysis of the entire distal femur in two different groups. Quantitative analysis of bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular number (Tb.N) and trabecular thickness (Tb.Th) of cancellous bone. (E) Immunohistochemical staining confirms that the expression of integrin β3, RUNX2, PDLIM5 and serpin E2 is significantly reduced in the bone tissues of integrin β3 KO mice compared with those from WT mice. Scale bar = 20 μm. (F) Tissue immunofluorescence staining analysis confirms the reduced expression of integrin β3, RUNX2, PDLIM5 and serpin E2 in the bone tissues of integrin β3 KO mice compared with those from WT mice. Scale bar = 20 μm. (G) Western blotting analysis demonstrates the expression levels of OPN, RUNX2, integrin β3, serpin E2, PDLIM5, α‐actinin 1 and lamin A in control and PDLIM5‐overexpressing hASCs under cyclic stretch loading. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. (H) Western blotting analysis demonstrates the expression levels of OPN, RUNX2, integrin β3, serpin E2, PDLIM5, α‐actinin 1 and lamin A in control and serpin E2‐overexpressing hASCs under cyclic stretch loading. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: In Vivo, Western Blot, Expressing, Derivative Assay, Isolation, Knock-Out, Staining, Micro-CT, Immunohistochemical staining, Immunofluorescence, Control

Journal: Cell Proliferation

Article Title: Mechanotransduction Mediated by PDLIM5 : The Critical Role of Serpin E2 /Integrin β3‐Cytoskeleton‐Nucleoskeleton Axis in Mechanical Osteogenic Programming

doi: 10.1111/cpr.70067

Figure Lengend Snippet: The schematic diagram of the regulatory mechanism illustrates that PDLIM5 mediates mechanical stimulation, thereby promoting osteogenic differentiation of human adipose‐derived stem cells (hASCs) via the serpin E2/integrin β3‐PDLIM5‐lamin A signalling axis.

Article Snippet: When the purified hASCs or mASCs reached 80% confluence, the GM was replaced with the osteogenic induction medium (OS) containing DMEM medium, 5% FBS, 1% penicillin/streptomycin, 100 nM dexamethasone (Sigma‐Aldrich), 37.5 mg/L ascorbic acid (Sigma‐Aldrich), 10 mM β‐glycerophosphate (Sigma‐Aldrich) and 10 nM VitD3 (Solarbio, Beijing, China).

Techniques: Derivative Assay

Journal: Development (Cambridge, England)

Article Title: BMP signaling orchestrates a transcriptional network to control the fate of mesenchymal stem cells in mice

doi: 10.1242/dev.150136

Figure Lengend Snippet: Gli1 is an in vivo marker for MSCs in the developing molar apical mesenchyme. (A-D) X-gal staining (blue) of sagittal sections of mandibular molars from Gli1-LacZ mice at P0.5, P3.5, P7.5 and P21.5. Arrowheads indicate Gli1+ cells in the apical region of the mesenchyme. (E-H) Visualization of sagittal sections of mandibular molars from Gli1-CreER;tdTomato mice at P4.5 and P18.5 after induction at P3.5. The progeny of the Gli1 lineage appear red. Boxes in E and G are shown at higher magnification in F and H, respectively. (I-L) Schematic drawing of Gli1+ cells (dark blue, at the base of the developing root) and their progeny (light blue) contribute to root mesenchyme growth during root initiation (I,J), elongation (K) and eruption (L) stages. (M-P) Colony-forming assay and osteogenic, chondrogenic and adipogenic differentiation assays of cells from the Gli1+ region in the apical mesenchyme of molars from P5.5 Gli1-CreER;tdTomato mice induced at P3.5. Toluidine Blue staining was used to visualize colony formation after culture for 2 weeks (M). Inset shows that colonies are derived from Gli1+ cells (red). Alizarin Red (N), Alcian Blue (O) and Oil Red O (P) staining to detect osteogenic, chondrogenic and adipogenic differentiation after 3 weeks. Scale bars: 100 μm.

Article Snippet: For the differentiation assays, cells from the colonies were cultured until confluent and then induced in osteogenic, adipogenic or chondrogenic differentiation medium (05504, 05503 and 05455, Stemcell Technologies) for 2-3 weeks according to the manufacturer's instructions.

Techniques: In Vivo, Marker, Staining, Derivative Assay

Journal: Frontiers in Medicine

Article Title: Dual-functional sulfonated PEEK implants via graphene oxide–mediated BMP-2 gene delivery: enhanced osteogenic and antibacterial performance in vitro

doi: 10.3389/fmed.2026.1763692

Figure Lengend Snippet: Osteogenic properties of SPEEK-(GO-PEG/pBMP-2). (A) ALP staining of rBMSCs cultured on different substrates. (B) Quantitative analysis of ALP activity. (C) Alizarin Red S staining of mineralized nodules. (D) Semi-quantitative analysis of mineral deposition. (E–H) qRT-PCR analysis of osteogenesis-related genes: (E) COL-1, (F) ALP, (G) Runx2, and (H) OPN. (ns, p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: Cells were cultured in osteogenic induction medium (Cat. No. PD-008, Procell, Wuhan, China) for 7 and 14 days, respectively.

Techniques: Staining, Cell Culture, Activity Assay, Quantitative RT-PCR

Journal: Macromolecular Bioscience

Article Title: NIR‐Responsive ZIF‐8 Metal‐Organic Framework Nanohybrids with Photothermal, Antimicrobial, and Osteoinductive Properties to Prevent Implant Infection

doi: 10.1002/mabi.202400594

Figure Lengend Snippet: Photothermal antibacterial activity of Au@ZIF‐8 nanohybrid. A) Illustration depicting the antibacterial activation mechanism and osteogenesis stimulation process on Au@ZIF‐8‐coated titanium substrate under NIR irradiation. NIR irradiation triggers the photothermal conversion of the Au@ZIF‐8 nanohybrid, causing localized heating and ZIF‐8 degradation. This process enhances antibacterial effects by disrupting bacterial cell membranes. Simultaneously, the released zinc ions from the degraded ZIF‐8 promote osteogenic differentiation of surrounding cells, supporting bone tissue regeneration and mineralization on the titanium substrate. B) The optical images compare the Titanium implant before and after coating with Au@ZIF‐8, demonstrating a noticeable visual difference indicative of successful nanoparticle deposition. C) Thermal images of the Au@ZIF‐8‐coated Titanium implant under 5 min of NIR irradiation reveal a significant temperature increase, confirming the photothermal properties of the Au@ZIF‐8 coating. D) Temperature changes after 5 min of NIR irradiation and subsequent 5 min of cooling demonstrate that the Au@ZIF‐8 coated Titanium implant retained heat more effectively, suggesting their potential for enhanced therapeutic activity. E) and F) SEM images of Au@ZIF‐8‐coated titanium implants, without and with PDA coating respectively. G) and H) display the corresponding EDX analyses graph exhibiting distinctive Zn (coming from Au@ZIF‐8) and Ti peaks (coming from titanium implant). I) Photographs of MRSA bacterial colonies treated with a PBS blank, ZIF‐8, and varying concentrations of Au@ZIF‐8 nanohybrids were analyzed using the plate count method, both under NIR irradiation for 30 min and without light exposure in the dark. Au@ZIF‐8 nanohybrids demonstrated increased antibacterial effectiveness with higher concentrations. J) The relative bacterial viabilities were measured following treatment with a PBS blank, ZIF‐8, and Au@ZIF‐8, both under 30 min of NIR irradiation and in the dark. The PBS control in the dark was used as a reference for 100% cell viability. A value of **** p < 0.0001 indicated a highly significant difference ( n = 3).

Article Snippet: MC3T3 cells were seeded onto ZIF‐8 and Au@ZIF‐8 nanohybrids encapsulated within a GelMA hydrogel and cultured in an osteogenic induction medium for 21 days.

Techniques: Activity Assay, Activation Assay, Irradiation, Control

Journal: Macromolecular Bioscience

Article Title: NIR‐Responsive ZIF‐8 Metal‐Organic Framework Nanohybrids with Photothermal, Antimicrobial, and Osteoinductive Properties to Prevent Implant Infection

doi: 10.1002/mabi.202400594

Figure Lengend Snippet: Osteogenic differentiation and In vitro cytocompatibility of Au@ZIF‐8 nanoparticles in hydrogels. A) Schematic visualization of the method by which the cells were exposed to the Au@ZIF‐8 hydrogels. B) Bright field and fluorescence live/dead cell images of pre‐osteoblast cells, showing excellent cell biocompatibility. C) Cell viability of pre‐osteoblast cells evaluated by MTS cell proliferation assay, showing no adverse effect on cells when Au@ZIF‐8 in the nanohybrid hydrogels ( n = 5). D) Alizarin Red staining was used to compare the Ctrl, Au@ZIF‐8 (‐), and Au@ZIF‐8 (+), revealing increased mineral deposition in the Au@ZIF‐8‐treated groups. E) The corresponding ARS optical density value at 405 nm analysis revealed a significant increase in calcium deposition in the Au@ZIF‐8‐treated groups. F) RT‐qPCR analysis revealed that the presence of ZIF‐8 significantly upregulated osteogenic‐related genes, including OCN, OPN, Runx2, and BMP2, underscoring its effectiveness in promoting bone regeneration. A value of * p > 0.05 indicated no significant difference, a value of ** p < 0.01 indicated a significant difference, *** p < 0.001 **** p < 0.0001 indicated a highly significant difference. ( n = 3).

Article Snippet: MC3T3 cells were seeded onto ZIF‐8 and Au@ZIF‐8 nanohybrids encapsulated within a GelMA hydrogel and cultured in an osteogenic induction medium for 21 days.

Techniques: In Vitro, Fluorescence, Proliferation Assay, Staining, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Extracellular Vesicles Derived from Human Umbilical Cord Mesenchymal Stem Cells Alleviated the Inflammatory Response in Mice Infected with the Influenza Virus A (H1N1)

doi: 10.3390/ijms26188839

Figure Lengend Snippet: Characterization of human umbilical cord mesenchymal stem cells (hUCMSCs) and extracellular vesicles derived from hUCMSCs (hUCMSC-EVs). ( A ) hUCMSCs passed to the 3rd generation grew adherently to the wall and were in good condition, with the morphology of elongated pike/pike shape and the overall distribution of vortex (Scale bar = 200 μm). ( B ) Oil red O staining of hUCMSCs after lipogenic induction, visible with obvious red lipid droplets (Scale bar = 100 μm). ( C ) Alizarin red staining of hUCMSCs after osteogenic induction, visible with obvious red calcium nodules (Scale bar = 100 μm). ( D ) Identification of hUCMSCs surface markers by flow cytometry. ( E ) ZETAVIEW particle potential titration and particle size analyzer for hUCMSC-EVs particle size detection. ( F ) Transmission electron microscopy to observe hUCMSC-EVs morphology (scale bar = 200 nm), the red arrow indicates the hUCMSC-EVs. ( G ) Western blotting to identify the expression of hUCMSC-EVs marker protein.

Article Snippet: When the cell density reached 80–90%, the cells were cultured in adipogenic differentiation induction medium (Dakewe, Beijing, China, #6114531) and osteogenic differentiation induction medium (Dakewe, Beijing, China, #6114541) for 18–21 days.

Techniques: Derivative Assay, Staining, Flow Cytometry, Titration, Transmission Assay, Electron Microscopy, Western Blot, Expressing, Marker