Journal: Molecules

Article Title: Natural Polyphenol Corilagin Enhances Osteogenesis and Chondrogenesis Differentiation of Mesenchymal Stem Cells: Implications for Bone and Cartilage Regeneration

doi: 10.3390/molecules31010194

Figure Lengend Snippet: Representative cell morphology of BM-MSCs at the third passage as observed under a light microscope at 4× magnification.

Article Snippet: Primary Bone Marrow-Derived Mesenchymal stem cells from normal human (BM-MSCs, ATCC ® number PCS-500-012TM, Lot: 70017526) and human synovial sarcoma cell line (SW-982, HTB-93 TM , Lot: 63971143) were purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Light Microscopy

Journal: Molecules

Article Title: Natural Polyphenol Corilagin Enhances Osteogenesis and Chondrogenesis Differentiation of Mesenchymal Stem Cells: Implications for Bone and Cartilage Regeneration

doi: 10.3390/molecules31010194

Figure Lengend Snippet: ( A ) Alizarin red S staining revealed the presence of calcium deposits 21 days after the induction of osteogenic medium (OM) in the presence of CLG at varying concentrations (1 μg/mL, 5 μg/mL, and 10 μg/mL). The scale bar (⎯) represents 200 µm. ( B ) Pellet cultures of BM-MSCs were sectioned and stained with H&E, showing the morphology at 21 days after the induction of chondrogenic medium (CM) with CLG at varying concentrations (1 μg/mL, 5 μg/mL, and 10 μg/mL). The scale bar (⎯) represents 200 µm. ( C ) Pellet cultures of BM-MSCs were sectioned and stained with toluidine blue, showing glycosaminoglycans at 21 days after the induction of chondrogenic medium (CM) with CLG at varying concentrations (1 μg/mL, 5 μg/mL, and 10 μg/mL). The scale bar (⎯) represents 200 µm. ( D ) Pellet cultures of BM-MSCs were sectioned and stained with Alcian blue, showing proteoglycans at 21 days after the induction of chondrogenic medium (CM) with CLG at varying concentrations (1 μg/mL, 5 μg/mL, and 10 μg/mL). The scale bar (⎯) represents 200 µm.

Article Snippet: Primary Bone Marrow-Derived Mesenchymal stem cells from normal human (BM-MSCs, ATCC ® number PCS-500-012TM, Lot: 70017526) and human synovial sarcoma cell line (SW-982, HTB-93 TM , Lot: 63971143) were purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Staining

Journal: Molecules

Article Title: Natural Polyphenol Corilagin Enhances Osteogenesis and Chondrogenesis Differentiation of Mesenchymal Stem Cells: Implications for Bone and Cartilage Regeneration

doi: 10.3390/molecules31010194

Figure Lengend Snippet: ( A ) The effects of different doses of corilagin (1, 5 and 10 µg/mL) on osteogenic differentiation potentials (Runx2, OPN) of BM-MSCs on day 21. Gene expression of differentiation potentials was assessed using real-time PCR. The bar means SD. Data is shown as mean ± SD. Statistical significance was calculated using a one-way ANOVA test and significance is represented on graphs as * p -value < 0.05. ( B ) The effects of different doses of corilagin (1, 5 and 10 µg/mL) on chondrogenic differentiation potentials (Sox9, Aggrecan) of BM-MSCs on day 21. Gene expression of differentiation potentials was assessed using real-time PCR. The bar means SD. Data is shown as mean ± SD. Statistical significance was calculated using a one-way ANOVA test and significance is represented on graphs as * p -value < 0.05.

Article Snippet: Primary Bone Marrow-Derived Mesenchymal stem cells from normal human (BM-MSCs, ATCC ® number PCS-500-012TM, Lot: 70017526) and human synovial sarcoma cell line (SW-982, HTB-93 TM , Lot: 63971143) were purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Serum DLX6-AS1 and miR-141-3p as early diagnostic biomarkers and regulators of delayed fracture healing (DFH)

doi: 10.1186/s13018-025-06478-5

Figure Lengend Snippet: Interaction between DLX6-AS1 and miR-141-3p. A Predicted target sites. B Dual-luciferase reporter validation of the interaction. C Correlation analysis of serum DLX6-AS1 and miR-141-3p levels. D , E Expression of chondrogenic differentiation markers D , DLX6-AS1, and miR-141-3p E during hBMSCs chondrogenic differentiation. Data are presented as mean ± SD. Statistical analysis was performed using Student’s t-test or one-way ANOVA (n = 3). hBMSCs, Human bone marrow mesenchymal stem cells. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human bone marrow mesenchymal stem cells (hBMSCs) were obtained from ATCC (USA).

Techniques: Luciferase, Biomarker Discovery, Expressing

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Serum DLX6-AS1 and miR-141-3p as early diagnostic biomarkers and regulators of delayed fracture healing (DFH)

doi: 10.1186/s13018-025-06478-5

Figure Lengend Snippet: Effect of DLX6-AS1/miR-141-3p on hBMSCs functions. A qRT-PCR analysis of DLX6-AS1 and miR-141-3p after si-DLX6-AS1 treatment with or without miR-inhibition. B Proliferation level. C Apoptosis rate. D Expression of chondrogenic differentiation markers SOX9, COL2A1, and ACAN. Data are presented as mean ± SD. Statistical analysis was performed using Student’s t-test or one-way ANOVA (n = 3). hBMSCs, Human bone marrow mesenchymal stem cells. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human bone marrow mesenchymal stem cells (hBMSCs) were obtained from ATCC (USA).

Techniques: Quantitative RT-PCR, Inhibition, Expressing

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Serum DLX6-AS1 and miR-141-3p as early diagnostic biomarkers and regulators of delayed fracture healing (DFH)

doi: 10.1186/s13018-025-06478-5

Figure Lengend Snippet: VEGFA is a binding target of miR-141-3p. A Predicted binding sequence. B Dual-luciferase assay validation. C VEGFA expression levels in NFH (n = 96) vs DFH (n = 96) patients. D , E Correlation analyses between VEGFA and miR-141-3p or DLX6-AS1 (n = 96). Data are presented as mean ± SD. Statistical analysis was performed using Student’s t-test or one-way ANOVA. hBMSCs, Human bone marrow mesenchymal stem cells. NFH, normal fracture healing; DFH, delayed fracture healing. ** P < 0.01, *** P < 0.001

Article Snippet: Human bone marrow mesenchymal stem cells (hBMSCs) were obtained from ATCC (USA).

Techniques: Binding Assay, Sequencing, Luciferase, Biomarker Discovery, Expressing

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Serum DLX6-AS1 and miR-141-3p as early diagnostic biomarkers and regulators of delayed fracture healing (DFH)

doi: 10.1186/s13018-025-06478-5

Figure Lengend Snippet: Functional role of the miR-141-3p/VEGFA axis in hBMSCs. A miR-141-3p and VEGFA abundance after transfection. B Proliferation assessed. C Apoptosis analysis. D Expression of chondrogenic differentiation markers after indicated treatments. Data are presented as mean ± SD. Statistical analysis was performed using Student’s t-test or one-way ANOVA (n = 3). hBMSCs, Human bone marrow mesenchymal stem cells. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human bone marrow mesenchymal stem cells (hBMSCs) were obtained from ATCC (USA).

Techniques: Functional Assay, Transfection, Expressing

Journal: International Journal of Molecular Sciences

Article Title: BGN Secreted by Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression via Activation of TLR4-Mediated Erk and NF-κB Signaling Pathways

doi: 10.3390/ijms262412024

Figure Lengend Snippet: Biglycan (BGN), highly expressed in cancer-associated fibroblasts (CAFs), promotes the proliferation and migration of esophageal squamous cell carcinoma (ESCC) cells. ( A ) Euler diagram showing the overlap between genes meeting the signal intensity value criteria (CAF9 > 1000, TE-9 mono ≤ 100, and TE-9 co ≤ 100) and the expression ratio criteria (CAF9/MSC mono > 2 and TE-9 co/TE-9 mono < 2) in the cDNA microarray analysis ( GSE244020 and GSE274064 ). Eight overlapping genes were identified, with BGN showing the highest signal intensity value in CAF9. ( B ) Violin plot showing BGN expression across cell types based on single-cell RNA sequencing (scRNAseq) data from the GSE160269 dataset. ( C ) Uniform Manifold Approximation and Projection (UMAP) and BGN expression in fibroblasts based on scRNAseq data. Violin plot visualization shows significantly higher BGN expression in CAFs compared to normal fibroblasts. ( D – F ) The mRNA expression, protein expression, and secreted protein levels of BGN in mesenchymal stem cell (MSC) mono and CAFs were compared using quantitative reverse transcription polymerase chain reaction (qRT-PCR) ( D ) and Western blotting of cell lysates and supernatants ( E , F ), respectively. ( G ) The effects of recombinant human BGN (rhBGN) (100 ng/mL) on the proliferation of ESCC cells were evaluated using the MTS assay. ( H , I ) The effect of rhBGN (100 ng/mL) on ESCC cell migration ( H ) and the effect of BGN neutralizing antibody (220 ng/mL) on CAF-mediated ESCC cell migration ( I ) were assessed using the Transwell migration assay. In ( H ), ESCC cells were seeded in the upper chamber; in ( I ), ESCC cells were seeded in the upper chamber, while direct co-culture of ESCC cells and MSCs was performed in the lower chamber. The rhBGN ( H ) and BGN neutralizing antibody (220 ng/mL) or control immunoglobulin G (IgG; 220 ng/mL) ( I ) were added to the lower chamber. Migrated cells were counted in five representative microscopic fields after 48 h, and representative images are shown below the graphs. The data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( D , G – I ). * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bars: 100 μm ( H , I ). FRC, fibroblastic reticular cells.

Article Snippet: Human bone marrow-derived MSCs (PCS-500-012, ATCC, Manassas, VA, USA) were cultured in low-glucose Dulbecco’s Modified Eagle Medium (DMEM; FUJIFILM Wako Pure Chemical Corporation) containing 10% FBS and 1% antibiotics.

Techniques: Migration, Expressing, Microarray, RNA Sequencing, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Recombinant, MTS Assay, Transwell Migration Assay, Co-Culture Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: BGN Secreted by Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression via Activation of TLR4-Mediated Erk and NF-κB Signaling Pathways

doi: 10.3390/ijms262412024

Figure Lengend Snippet: Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein (FAP) and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained unchanged. TLR4 expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).

Article Snippet: Human bone marrow-derived MSCs (PCS-500-012, ATCC, Manassas, VA, USA) were cultured in low-glucose Dulbecco’s Modified Eagle Medium (DMEM; FUJIFILM Wako Pure Chemical Corporation) containing 10% FBS and 1% antibiotics.

Techniques: Migration, Activation Assay, Expressing, RNA Sequencing, MTS Assay, Transwell Migration Assay, Recombinant, Western Blot, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: BGN Secreted by Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression via Activation of TLR4-Mediated Erk and NF-κB Signaling Pathways

doi: 10.3390/ijms262412024

Figure Lengend Snippet: A schematic illustration of the role of BGN in the tumor–stromal interactions among ESCC cells, CAFs, and macrophages. BGN secreted from MSCs that have undergone CAF transition upon direct contact with ESCC cells promotes ESCC cell proliferation and migration through the TLR4–Erk/NF-κB signaling pathways. In addition, BGN enhances the proliferation and migration of MSCs and induces their differentiation into CAFs. Furthermore, BGN promotes the proliferation and migration of macrophages and drives their polarization toward the M2 phenotype through the TLR4–NF-κB signaling pathways.

Article Snippet: Human bone marrow-derived MSCs (PCS-500-012, ATCC, Manassas, VA, USA) were cultured in low-glucose Dulbecco’s Modified Eagle Medium (DMEM; FUJIFILM Wako Pure Chemical Corporation) containing 10% FBS and 1% antibiotics.

Techniques: Migration, Protein-Protein interactions

Journal: Journal of Materials Science. Materials in Medicine

Article Title: Copper tannic acid coordination nanosheet as a potent in-situ antibiotic sustained-release carrier for chronic osteomyelitis

doi: 10.1007/s10856-025-06979-z

Figure Lengend Snippet: The effect of Van@CuTA on BMSCs and HUVECs. A CCK-8 assay demonstrated no toxicity of Van@CuTA toward BMSCs. B Representative images of alizarin red staining, revealing increased calcium deposition in BMSCs treated with Van@CuTA. C , D Representative images and statistical analysis of HUVECs migration assay demonstrating a significant increase in migrated cells following Van@CuTA treatment. E – G Tube formation assay images and corresponding quantifications of total tube length and node number per view indicated that Van@CuTA significantly promoted HUVECs’ tube formation. (* indicated p < 0.05)

Article Snippet: Primary bone marrow mesenchymal stem cells (BMSCs) and Human Umbilical Vein Endothelial Cells (HUVECs) were obtained from ATCC.

Techniques: CCK-8 Assay, Staining, Migration, Tube Formation Assay