Journal: Bioactive Materials

Article Title: A “built-up” composite film with synergistic functionalities on Mg–2Zn–1Mn bioresorbable stents improves corrosion control effects and biocompatibility

doi: 10.1016/j.bioactmat.2023.02.004

Figure Lengend Snippet: (A) Fluorescent staining of HUVECs cultured for 1 day, 3 days, and 5 days. (B) HUVECs' viability after 1 day, 3 days, and 5 days of incubation is calculated from CCK-8 tests. (C) Fluorescent staining of HUASMCs cultured for 1 day, 3 days, and 5 days. (D) HUASMCs' viability after 1 day, 3 days, and 5 days of incubation is calculated from CCK-8 tests. Data are presented as mean ± SD (n = 4), *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: In the cell culture experiment, human umbilical vein endothelial cells (HUVECs) and human umbilical artery smooth muscle cells (HUASMCs) purchased from Guangzhou Geneo Biotech were first subcultured and then seeded in 24-well plates at a density of 1.5 × 10 4 and 2 × 10 4 cells/ml, respectively.

Techniques: Staining, Cell Culture, Incubation, CCK-8 Assay

Journal: Stem Cell Research & Therapy

Article Title: TGFβ3 recruits endogenous mesenchymal stem cells to initiate bone regeneration

doi: 10.1186/s13287-017-0693-0

Figure Lengend Snippet: Transwell assay for hBMSC migration. a Analysis for the migration of hBMSCs with or without hUVECs and hUASMCs. b In the coculture system of hBMSCs and hUASMCs, representative light photomicrographs of migrated hBMSC induced by 0-100 ng/ml TGFβ3 after 24-hour incubation. c In the coculture system of hBMSCs and hUVECs, representative light photomicrographs of migrated hBMSCs induced by 0-100 ng/ml TGFβ3 after 24-hour incubation. d Quantitative analysis of migrated cell density for ( c ) and ( d ). Migrated cells were stained purple with crystal violet . Scale bar: 100 μm. * P < 0.05, ** P < 0.01, *** P < 0.005, *** P < 0.001. hUASMC human umbilical artery smooth muscle cell, hUVEC human umbilical vein endothelial cell, MSC mesenchymal stem cell, TGFβ3 transforming growth factor beta-3

Article Snippet: Human umbilical artery smooth muscle cells (hUASMCs) and human umbilical vein endothelial cells (hUVECs) were from ScienCell (CA, USA). hUASMCs were cultured in smooth muscle cell medium (ScienCell Inc., CA, USA), and hUVECs were cultured in endothelial cell medium (ScienCell Inc., CA, USA).

Techniques: Transwell Assay, Migration, Incubation, Staining

Journal: Stem Cell Research & Therapy

Article Title: TGFβ3 recruits endogenous mesenchymal stem cells to initiate bone regeneration

doi: 10.1186/s13287-017-0693-0

Figure Lengend Snippet: TGFβ3 upregulated MCP1 secretion from vascular cells by TβRII/Smad3 signaling. a Secretion of MCP1 and SDF1 from vascular cells in the coculture system hBMSCs and vascular cells with TGFβ3. b Secretion of MCP1 from hBMSCs, hUVECs, and hUASMCs in different systems. c Expression of TβRII in hUVECs and hUASMCs induced by TGFβ3 assessed by western blot analysis. d Expression of p-Smad3 and Smad3 in hUVECs and hUASMCs induced by TGFβ3 assessed by western blot analysis. ** P < 0.01, *** P < 0.005, **** P < 0.001. hUASMC human umbilical artery smooth muscle cell, hUVEC human umbilical vein endothelial cell, MSC mesenchymal stem cell, TGFβ3 transforming growth factor beta-3

Article Snippet: Human umbilical artery smooth muscle cells (hUASMCs) and human umbilical vein endothelial cells (hUVECs) were from ScienCell (CA, USA). hUASMCs were cultured in smooth muscle cell medium (ScienCell Inc., CA, USA), and hUVECs were cultured in endothelial cell medium (ScienCell Inc., CA, USA).

Techniques: Expressing, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: TGFβ3 recruits endogenous mesenchymal stem cells to initiate bone regeneration

doi: 10.1186/s13287-017-0693-0

Figure Lengend Snippet: Knockdown of Smad3 in vascular cells inhibited TGFβ3-induced hBMSC migration. a Expression of Smad3 in hUVECs and hUASMCs were transfected with siRNA Smad3 as assessed by western blot analysis. b Relative density of Smad3 for ( a ). c Secretion of MCP1 in different cells. d Transwell assay for hBMSC migration in the coculture system of hBMSC and vascular cells with or without knockdown of Smad3. Migrated cells were stained purple with crystal violet. Scale bar: 100 μm. ** P < 0.01, **** P < 0.001. hUASMC human umbilical artery smooth muscle cell, hUVEC human umbilical vein endothelial cell, MSC mesenchymal stem cell, siRNA small interfering RNA, TGFβ3 transforming growth factor beta-3

Article Snippet: Human umbilical artery smooth muscle cells (hUASMCs) and human umbilical vein endothelial cells (hUVECs) were from ScienCell (CA, USA). hUASMCs were cultured in smooth muscle cell medium (ScienCell Inc., CA, USA), and hUVECs were cultured in endothelial cell medium (ScienCell Inc., CA, USA).

Techniques: Knockdown, Migration, Expressing, Transfection, Western Blot, Transwell Assay, Staining, Small Interfering RNA

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: The expression of RXFP1 and RXFP2 receptor mRNA and RXFP1 receptor protein in human primary umbilical vascular cells and human primary cardiac fibroblasts. qPCR (A) was utilized to show expression levels of RXFP1 and RXFP2 receptor mRNA in HUAECs, HUVECs, HUASMCs, HUVSMCs and HCFs relative to β-actin (n = 2). RXFP2 receptor mRNA was only measureable in the positive control. Cell surface RXFP1 receptor protein expression was determined by radioligand binding (B) utilizing [125I]-serelaxin and showed specific serelaxin binding in HEK-RXFP1 cells (n = 6), HUASMCs (n = 4), HUVECs (n = 4), HUVSMCs (n = 3) and HCFs (n = 3), but not in HUAECs (n = 2).

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Expressing, Positive Control, Binding Assay

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: The effect of serelaxin on cAMP accumulation in human primary umbilical vascular cells and cardiac fibroblasts. Serelaxin treatment (30 min) increased cAMP accumulation in (A) HUVECs (n = 6), (B) HUVSMCs (n = 6) and (C) HUASMCs (n = 4), but not in (D) HCFs (n = 3). The serelaxin CRC was bell-shaped for HUVECs and HUVSMCs but sigmoidal for HUASMCs. For each cell type, the effect of PTX (50 ng·mL−1, 18 h) and wortmannin (100 nM, 30 min) pretreatment was determined after exposure to serelaxin (30 nM) for 30 min to determine the role of Gαi and PI3K. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with serelaxin alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: The effect of serelaxin on cGMP accumulation in human primary umbilical vascular cells and cardiac fibroblasts. Serelaxin treatment (30 min) increased cGMP accumulation in (A) HUVECs (n = 7), (B) HUVSMCs (n = 5), (C) HUASMCs (n = 6) and (D) HCFs (n = 5). The serelaxin CRC was bell-shaped for HUVECs, HUVSMCs and HCFs but sigmoidal for HUASMCs. PTX (50 ng·mL−1, 18 h) and wortmannin (100 nM, 30 min) pretreatment significantly inhibited serelaxin (30 nM)-mediated cGMP accumulation in each cell type. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with serelaxin alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: Changes in the expression of nNOS, VEGF and ETB receptors in human primary umbilical vascular cells and cardiac fibroblasts after serelaxin (1.68 nM) exposure for 24 and 48 h. In HUVECs (A), serelaxin treatment increased the expression of nNOS (n = 5), ETB receptors (n = 6) and VEGF (n = 7). In HUVSMCs (B), serelaxin treatment increased the expression of nNOS (n = 5) and ETB (n = 5) but not VEGF (n = 4); however, in HUASMC (C), serelaxin treatment increased the expression of nNOS (n = 6), ETB receptors (n = 7) and VEGF (n = 5). In HCFs (D), similar to HUVECs and HUASMCs, serelaxin treatment increased the expression of nNOS (n = 5), ETB receptors (n = 5) and VEGF (n = 5). A representative blot of each protein and β-actin, a loading control, is shown with the densitometry in each figure. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with vehicle alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Expressing, Control

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: Changes in the activity of MMPs in human primary umbilical vascular cells and cardiac fibroblasts using zymography to assess changes in activity of MMP2 and MMP9 after long-term serelaxin exposure (1.68 and 30 nM) for 48 h. Serelaxin treatment increased the activity of MMP2 in (A) HUVECs (n = 5), (B) HUVSMCs (n = 6), (C) HUASMCs (n = 7) and (D) HCFs (n = 5). Serelaxin also significantly increased the activity of MMP9 but only in (B) HUVSMCs (n = 5) and (D) HCFs (n = 5) and not in (A) HUVECs (n = 5) and (C) HUASMCs (n = 5). Furthermore, and consistent with the lack of RXFP1 receptor expression (Figure 1), serelaxin had no effect on MMP activity in HUAECs (n = 2). A representative scan of each zymography is shown along with the densitometry in each figure. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with control alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Activity Assay, Zymography, Expressing, Control

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: Signal transduction mechanisms employed by serelaxin in human umbilical vascular cells and HCFs and their potential physiological effects. Short-term (<1 h) serelaxin stimulation produces cAMP/cGMP accumulation in vascular cells and pERK1/2 in all cells that is differentially regulated by Gαs, Gαi, GαOB and PI3K, and these pathways are likely to be involved in the vasodilator and anti-apoptotic effects of serelaxin respectively. In HUASMCs, the serelaxin-mediated cAMP or VEGF response did not involve GαOB and PI3K, whereas in HCFs, the RXFP1 receptor was not coupled to cAMP production. Longer term (24–48 h) serelaxin treatment increased VEGF expression involving both cAMP-dependent and cAMP-independent mechanisms, and these pathways are likely to be involved in angiogenesis. In addition, serelaxin treatment increased the activity of MMP2 and MMP9 to mediate its remodelling actions, but these enzymes are also secreted to convert big ET to ET1-32 that activates ETB receptors to further enhance vasodilatation (Conrad, 2010). Solid lines indicate mechanisms identified in the current study whereas dashed lines indicate previously established mechanisms.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Transduction, Expressing, Activity Assay