Journal: Journal of Orthopaedic Translation

Article Title: JD-312 – A novel small molecule that facilitates cartilage repair and alleviates osteoarthritis progression

doi: 10.1016/j.jot.2023.11.007

Figure Lengend Snippet: Identification of JD-312 as compound that promotes cartilage differentiation A) Schematic of drug screening strategy. B) JD-312 increases chondrocyte-specific gene expression in hUCMSCs (JD-312, 10 μM. KGN, 10 μM). Cells were incubated for 7 days to detect mRNA expression levels via RT-qPCR (n = 3). Type II Collagen, Col2a1 . Aggrecan, Acan . C) The protein levels of Col2 and ACAN in hUCMSCs were analysed by western blotting. D) Effect of JD-312 on cell viability of hUCMSCs detected by CCK8 assay (n = 3).

Article Snippet: We used a micromass culture of hUCMSCs to validate the effectiveness of JD-312 in inducing chondrogenic differentiation.

Techniques: Drug discovery, Gene Expression, Incubation, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay

Journal: Journal of Orthopaedic Translation

Article Title: JD-312 – A novel small molecule that facilitates cartilage repair and alleviates osteoarthritis progression

doi: 10.1016/j.jot.2023.11.007

Figure Lengend Snippet: JD-312 promotes the chondrogenic differentiation of hUCMSCs in vitro . A) Alcian blue staining showing the effect of JD-312 on chondrogenic differentiation of hUCMSCs. B) RT-qPCR analysis of Col2a1 and Acan mRNA expression in hUCMSCs treated with varying concentrations of JD-312. C) Alcian blue staining showing the effect of JD-312 (10 μM) on chondrogenic differentiation of hUCMSCs over time. D) RT-qPCR analysis of Col2a1 and Acan mRNA expression over time in hUCMSCs treated with JD-312. E) Macroscopic images of 3D micromass cultures of hUCMSCs treated with kartogenin (KGN) and JD-312. F) illustrates the weight of 3D micromass cultures shown in E. G) The concentration of glycosaminoglycan (GAG) released in the culture supernatant of the 3D micromass cultures at 21 days. (*P < 0.05, **P < 0.01, ***P < 0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: We used a micromass culture of hUCMSCs to validate the effectiveness of JD-312 in inducing chondrogenic differentiation.

Techniques: In Vitro, Staining, Quantitative RT-PCR, Expressing, Concentration Assay

Journal: Journal of Orthopaedic Translation

Article Title: JD-312 – A novel small molecule that facilitates cartilage repair and alleviates osteoarthritis progression

doi: 10.1016/j.jot.2023.11.007

Figure Lengend Snippet: JD-312 treated hUCMSCs enhances cartilage repair after DMM surgery in SD rats A) Representative images of Safranin-O staining of knee joints following sham operation or DMM in rats which were treated with hUCMSCs (MSC), or hUCMSCs transiently induced with KGN (KGN + MSC) or JD-312 (JD-312+MSC), with PBS as control. Scale bar = 400 μm. The OARSI scores for the sections are reported in B) . Knee joint sections were stained with antibodies against Col2 and ACAN and the expression is quantified in C) . n = 6 per group. Red, Col2a1, ACAN; Blue, DAPI. (*P < 0.05, **P < 0.01, ***P < 0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: We used a micromass culture of hUCMSCs to validate the effectiveness of JD-312 in inducing chondrogenic differentiation.

Techniques: Staining, Control, Expressing

Journal: Journal of Orthopaedic Translation

Article Title: JD-312 – A novel small molecule that facilitates cartilage repair and alleviates osteoarthritis progression

doi: 10.1016/j.jot.2023.11.007

Figure Lengend Snippet: Identifying the molecular mechanisms that govern JD-312's chondrogenic effect in hUCMSCS A) Volcano plots depicting the differentially expressed genes between hUCMSCs treated with JD-312 (10 μM) after 7 days. B and C) KEGG and GSEA enrichment analysis of the differentially expressed genes. D) Heatmap depicting the fold changes of differentially expressed genes associated with the Focal adhesion, PI3K-Akt, and ECM-receptor interaction pathways highlighted.

Article Snippet: We used a micromass culture of hUCMSCs to validate the effectiveness of JD-312 in inducing chondrogenic differentiation.

Techniques:

Journal: PLoS ONE

Article Title: Interleukin-1β-induced matrix metalloproteinase-3 via ERK1/2 pathway to promote mesenchymal stem cell migration

doi: 10.1371/journal.pone.0252163

Figure Lengend Snippet: (A) Cell wound healing assay for IL-1β stimulated mesenchymal stem cells in the presence or absence of 2 μg/ml IL-1RA (IL-1β inhibitor) at 12 and 24 hours. Scale bars = 300 μm. (B) The wound area of hUCMSCs were indicated by MetaMorph and the data were normalized with control and shown as the mean ± SD (n = 3, *P<0.05, **P<0.01 versus control cells, #p<0.05, ##p<0.01 versus IL-1β treated cells). (C) Cell viability assay for IL-1β stimulation. Data were quantified by multimode micro-plate readers. Data are shown as the mean ± SD (n = 3) (N.S.: nonsignificance).

Article Snippet: Human umbilical cord derived mesenchymal stem cells (hUCMSCs) were purchased from Bioresource Collection and Research Center, Hsinchu, Taiwan. hUCMSCs were cultured in low serum defined medium consisting of 56% low-glucose Dulbecco’s Modified Eagle Medium (DMEM-LG; Invitrogen, CA, USA), 37% MCBD 201 (Sigma, MO, USA), 2% fetal bovine serum (Thermo, Logan, UT), 0.5 mg/ml of AlbuMAX® I (Invitrogen, CA, USA), 1X insulin-transferrin-selenium-A (Invitrogen, CA, USA), 1X antibiotic antimycotic solution (Thermo, Logan, UT), 10 nM dexamethasone (Sigma, MO, USA), 50 nM L-ascorbic acid 2-phosphate (Sigma, MO, USA), 10 ng/ml of epidermal growth factor (PeproTech, NJ, USA), and 1 ng/ml of platelet-derived growth factor-BB (PeproTech, NJ, USA) at 37°C and 5% CO 2 .

Techniques: Wound Healing Assay, Control, Viability Assay

Journal: PLoS ONE

Article Title: Interleukin-1β-induced matrix metalloproteinase-3 via ERK1/2 pathway to promote mesenchymal stem cell migration

doi: 10.1371/journal.pone.0252163

Figure Lengend Snippet: Total RNA microarray based screening for the expression of MMPs with IL-1β stimulation for 24 hours in mesenchymal stem cells.

Article Snippet: Human umbilical cord derived mesenchymal stem cells (hUCMSCs) were purchased from Bioresource Collection and Research Center, Hsinchu, Taiwan. hUCMSCs were cultured in low serum defined medium consisting of 56% low-glucose Dulbecco’s Modified Eagle Medium (DMEM-LG; Invitrogen, CA, USA), 37% MCBD 201 (Sigma, MO, USA), 2% fetal bovine serum (Thermo, Logan, UT), 0.5 mg/ml of AlbuMAX® I (Invitrogen, CA, USA), 1X insulin-transferrin-selenium-A (Invitrogen, CA, USA), 1X antibiotic antimycotic solution (Thermo, Logan, UT), 10 nM dexamethasone (Sigma, MO, USA), 50 nM L-ascorbic acid 2-phosphate (Sigma, MO, USA), 10 ng/ml of epidermal growth factor (PeproTech, NJ, USA), and 1 ng/ml of platelet-derived growth factor-BB (PeproTech, NJ, USA) at 37°C and 5% CO 2 .

Techniques: Microarray, Expressing

Journal: PLoS ONE

Article Title: Interleukin-1β-induced matrix metalloproteinase-3 via ERK1/2 pathway to promote mesenchymal stem cell migration

doi: 10.1371/journal.pone.0252163

Figure Lengend Snippet: (A) Quantitation of changes in gene expression of MMP-3 detected by real-time PCR. hUCMSCs were transfected with MMP-3 siRNA and stimulated with IL-1β for 24 hours. (B) MMP-3 protein expression was measured using ELISA, transfected with MMP-3 siRNA and stimulated with IL-1β for 36 hours. (C) Cell wound healing assay for IL-1β stimulated mesenchymal stem cells after MMP-3 siRNA transfection. Scale bars = 300 μm. (D) The wound area of hUCMSCs were indicated by MetaMorph. (E) Cell invasion assay for IL-1β stimulated mesenchymal stem cells after MMP-3 siRNA transfection. Scale bars = 1 mm. (F) Graph indicates the invasion ability of stem cells. Data are shown as the mean ± SD (n = 3, **P<0.01, ***P<0.005 versus control cells, #p<0.05, ##p<0.01, ###p<0.005, versus IL-1β treated cells).

Article Snippet: Human umbilical cord derived mesenchymal stem cells (hUCMSCs) were purchased from Bioresource Collection and Research Center, Hsinchu, Taiwan. hUCMSCs were cultured in low serum defined medium consisting of 56% low-glucose Dulbecco’s Modified Eagle Medium (DMEM-LG; Invitrogen, CA, USA), 37% MCBD 201 (Sigma, MO, USA), 2% fetal bovine serum (Thermo, Logan, UT), 0.5 mg/ml of AlbuMAX® I (Invitrogen, CA, USA), 1X insulin-transferrin-selenium-A (Invitrogen, CA, USA), 1X antibiotic antimycotic solution (Thermo, Logan, UT), 10 nM dexamethasone (Sigma, MO, USA), 50 nM L-ascorbic acid 2-phosphate (Sigma, MO, USA), 10 ng/ml of epidermal growth factor (PeproTech, NJ, USA), and 1 ng/ml of platelet-derived growth factor-BB (PeproTech, NJ, USA) at 37°C and 5% CO 2 .

Techniques: Quantitation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Wound Healing Assay, Invasion Assay, Control

Journal: PLoS ONE

Article Title: Interleukin-1β-induced matrix metalloproteinase-3 via ERK1/2 pathway to promote mesenchymal stem cell migration

doi: 10.1371/journal.pone.0252163

Figure Lengend Snippet: (A) Quantitation of changes in gene expression of MMP-3 detected by real-time PCR after ERK1/2 inhibitor U0126 (10–30 μM) treatment and stimulation with IL-1β for 24 hours. (B) Example of Western blot results of the MMP-3 (54 kDa) from the lysates of cells treated with IL-1β and ERK1/2 inhibitor U0126 (20 μM). The full-length Western blots was showed in . (C) Quantitative graphs of the Western blot results of MMP-3 protein expression of (B). (D) MMP-3 protein expression was measured using ELISA, treated with U0126 (20 μM) and stimulated with IL-1β for 36 hours. (E) MMP-3 activity was measured by Fluorogenic peptide Assays. hUCMSCs treated with U0126 at concentration of 20 μM. (F) Cell wound healing assay for IL-1β stimulated mesenchymal stem cells in the presence of ERK1/2 inhibitor U0126 (20 μM) at 24 hours. Scale bars = 300 μm. (G) The wound area of hUCMSCs were indicated by MetaMorph and the data were normalized with control and shown as the mean ± SD (n = 3, ***P<0.01 versus control cells, ##p<0.01 versus IL-1β treated cells). (H) Cell wound healing assay for IL-1β stimulated mesenchymal stem cells in the presence of ERK1/2 inhibitor U0126 (20 μM) at 24 hours. Scale bars = 1 mm. (I) Graph indicates the invasion ability of stem cells. Data are shown as the mean ± SD (n = 3, *P<0.05, *** P <0.005 versus control cells, #p<0.05, ##p<0.01, ###p<0.005 versus IL-1β treated cells).

Article Snippet: Human umbilical cord derived mesenchymal stem cells (hUCMSCs) were purchased from Bioresource Collection and Research Center, Hsinchu, Taiwan. hUCMSCs were cultured in low serum defined medium consisting of 56% low-glucose Dulbecco’s Modified Eagle Medium (DMEM-LG; Invitrogen, CA, USA), 37% MCBD 201 (Sigma, MO, USA), 2% fetal bovine serum (Thermo, Logan, UT), 0.5 mg/ml of AlbuMAX® I (Invitrogen, CA, USA), 1X insulin-transferrin-selenium-A (Invitrogen, CA, USA), 1X antibiotic antimycotic solution (Thermo, Logan, UT), 10 nM dexamethasone (Sigma, MO, USA), 50 nM L-ascorbic acid 2-phosphate (Sigma, MO, USA), 10 ng/ml of epidermal growth factor (PeproTech, NJ, USA), and 1 ng/ml of platelet-derived growth factor-BB (PeproTech, NJ, USA) at 37°C and 5% CO 2 .

Techniques: Quantitation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Activity Assay, Concentration Assay, Wound Healing Assay, Control

Journal: PLoS ONE

Article Title: Interleukin-1β-induced matrix metalloproteinase-3 via ERK1/2 pathway to promote mesenchymal stem cell migration

doi: 10.1371/journal.pone.0252163

Figure Lengend Snippet: (A) Quantitation of changes in gene expression of MMP-3 detected by real-time PCR after p38 inhibitor (50 nM SB205380), Akt inhibitor (20 μM GSK690693), and JNK inhibitor (20 nM SP600125) treatment and stimulation with IL-1β for 24 hours. (B) Cell wound healing assay for IL-1β stimulated mesenchymal stem cells in the presence 50 nM SB205380, 20 μM GSK690693, and 20 nM SP600125 at 24 hours. Scale bars = 300 μm. (C) Graph indicates the migration ability of stem cells into the wound area. Data are shown as the mean ± SD (n = 3, *P<0.05, ***P<0.005 versus control cells, ###p<0.005 versus IL-1β treated cells).

Article Snippet: Human umbilical cord derived mesenchymal stem cells (hUCMSCs) were purchased from Bioresource Collection and Research Center, Hsinchu, Taiwan. hUCMSCs were cultured in low serum defined medium consisting of 56% low-glucose Dulbecco’s Modified Eagle Medium (DMEM-LG; Invitrogen, CA, USA), 37% MCBD 201 (Sigma, MO, USA), 2% fetal bovine serum (Thermo, Logan, UT), 0.5 mg/ml of AlbuMAX® I (Invitrogen, CA, USA), 1X insulin-transferrin-selenium-A (Invitrogen, CA, USA), 1X antibiotic antimycotic solution (Thermo, Logan, UT), 10 nM dexamethasone (Sigma, MO, USA), 50 nM L-ascorbic acid 2-phosphate (Sigma, MO, USA), 10 ng/ml of epidermal growth factor (PeproTech, NJ, USA), and 1 ng/ml of platelet-derived growth factor-BB (PeproTech, NJ, USA) at 37°C and 5% CO 2 .

Techniques: Quantitation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Wound Healing Assay, Migration, Control

Journal: International Journal of Ophthalmology

Article Title: Protective effects of human umbilical cord mesenchymal stem cells on retinal ganglion cells in mice with acute ocular hypertension

doi: 10.18240/ijo.2021.02.03

Figure Lengend Snippet: A: CM-Dil-labeled 3rd passage hucMSCs (×100); B: CM-Dil-labeled hucMSCs (×100) 7d later; C: CM-Dil-labeled hucMSCs membrane (×200).

Article Snippet: The 3 rd generation of hucMSCs line was obtained from the SALIAI Stemcell Science and Technology Co., Ltd. (Guangzhou, China).

Techniques: Labeling, Membrane

Journal: International Journal of Ophthalmology

Article Title: Protective effects of human umbilical cord mesenchymal stem cells on retinal ganglion cells in mice with acute ocular hypertension

doi: 10.18240/ijo.2021.02.03

Figure Lengend Snippet: A: Normal; B: AOH; C: hucMSCs; D: NS. GCL: Ganglion cell layer; IPL: Inner plexus layer, INL: Inner nuclear layer, OPL: Outer plexus layer, ONL: Outer nuclear layer. AOH could significantly reduce the thickness of the inner retina. Intravitreal injection of hucMSCs could relieve the damage of AOH. Intravitreal injection of NS had no therapeutic effect on damage of AOH. aP<0.05.

Article Snippet: The 3 rd generation of hucMSCs line was obtained from the SALIAI Stemcell Science and Technology Co., Ltd. (Guangzhou, China).

Techniques: Injection

Journal: International Journal of Ophthalmology

Article Title: Protective effects of human umbilical cord mesenchymal stem cells on retinal ganglion cells in mice with acute ocular hypertension

doi: 10.18240/ijo.2021.02.03

Figure Lengend Snippet: A: Normal; B: High eye pressure; C: NS; D: hucMSCs; E: Results showed that AOH model could lead to the obvious death of RGCs; injection of hucMSCs in vitreous cavity could reduce the mortality of RGCs (P<0.05); injection of NS had no effect on RGCs (P>0.05). aP<0.05; bP<0.01.

Article Snippet: The 3 rd generation of hucMSCs line was obtained from the SALIAI Stemcell Science and Technology Co., Ltd. (Guangzhou, China).

Techniques: Injection