Journal: Materials Today Bio

Article Title: Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells

doi: 10.1016/j.mtbio.2025.102154

Figure Lengend Snippet: Experimental setup for bone quadruple cultures with primary human OB, OCy, OC and endothelial cells. OB were isolated from human femoral heads or femoral condyle. PBMC were isolated from human buffy coat by density gradient centrifugation. HUVEC, on top of OB were seeded to the apical side of transwell insert membranes. OB embedded in collagen gel were transferred to the basal side of the transwell inserts. Constructs were placed in well plates with PBMC. Over 14 days, collagen gel embedded OB differentiate to OCys and OCs differentiate simultaneously from PBMC.

Article Snippet: HUVEC, purchased from Promocell, were expanded in endothelial cell growth medium (EGM, Promocell) until passage 3.

Techniques: Isolation, Gradient Centrifugation, Construct

Journal: Materials Today Bio

Article Title: Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells

doi: 10.1016/j.mtbio.2025.102154

Figure Lengend Snippet: OB and HUVEC on transwell insert membranes in two quadruple culture experiments under different cultivation conditions. (a) Fluorescence microscopic images. Cytoskeleton appears green (iFluor488 phalloidin), nuclei appear blue (DAPI) and CD31 appears magenta (Alexa Fluor 546). Scale bars represent 500 μm. (b) Scheme of OB + HUVEC in quadruple culture. (c) Gene expression of OB markers presented as ΔCt (each experiment n = 4, in total n = 8). (d) ALP activity of OB (each experiment n = 3, in total n = 6). (e) Gene expression of endothelial cell markers presented as ΔCt (each experiment n = 4, in total n = 8). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: HUVEC, purchased from Promocell, were expanded in endothelial cell growth medium (EGM, Promocell) until passage 3.

Techniques: Fluorescence, Gene Expression, Activity Assay

Journal: Materials Today Bio

Article Title: Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells

doi: 10.1016/j.mtbio.2025.102154

Figure Lengend Snippet: OB and HUVEC on transwell insert membranes in two quadruple culture experiments in comparison to OB monocultures or OB-HUVEC co-cultures. (a) Fluorescence microscopic images in quadruple culture. Cytoskeleton appears green (iFluor488 phalloidin), nuclei appear blue (DAPI) and CD31 appears magenta (Alexa Fluor 546). Scale bars represent 500 μm. (b) Alizarin red staining of OB in monoculture and quadruple culture. Scale bars represent 500 μm. (c) Gene expression of OB markers presented as fold change ± upper and lower limit normalized to OB monocultures (each experiment n = 4). (d) ALP activity of OB (each experiment n = 3). (e) Gene expression of endothelial cell markers presented as fold change ± upper and lower limit normalized to OB-HUVEC co-cultures (each experiment n = 4). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: HUVEC, purchased from Promocell, were expanded in endothelial cell growth medium (EGM, Promocell) until passage 3.

Techniques: Comparison, Fluorescence, Staining, Gene Expression, Activity Assay

Journal: International Journal of Nanomedicine

Article Title: Bosentan Delivery via Nano Metal-Organic Framework nanoMIL-89 Restores Vascular Homeostasis in Pulmonary Arterial Hypertension

doi: 10.2147/IJN.S535437

Figure Lengend Snippet: Angiogenic effects of Bos@nanoMIL-89 in a 3D co-culture model of HUVECs and HPASMCs. ( A ) Representative immunofluorescence images showing dose-dependent enhancement of endothelial tube formation. Endothelial cells were stained with anti-CD31 (green), and all cell membranes were labeled using a red cell mask; scale bar, 100 μm. ( B ) Quantitative analysis of endothelial and smooth muscle cell counts, total tube length, and number of junctions across increasing concentrations of Bos@nanoMIL-89. Nine fields were analyzed per well (n = 6–12 per condition). Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA; *p < 0.05 vs control.

Article Snippet: HUVECs were maintained in EGM-2 medium (PromoCell, C-22011), and HPASMCs in high-glucose DMEM (Capricorn, DMEM-H-12), both supplemented with 10% fetal bovine serum (FBS; ScienCell, 0500), 1% penicillin/streptomycin (Gibco, 15140122), and 1% endothelial cell growth supplement for HUVECs only (PromoCell, C-39215).

Techniques: Co-Culture Assay, Immunofluorescence, Staining, Labeling, Control

Journal: International Journal of Nanomedicine

Article Title: Bosentan Delivery via Nano Metal-Organic Framework nanoMIL-89 Restores Vascular Homeostasis in Pulmonary Arterial Hypertension

doi: 10.2147/IJN.S535437

Figure Lengend Snippet: Angiogenic effects of nanoMIL-89 at various concentrations in a 3D co-culture model of HUVECs and HPASMCs. ( A ) Representative immunofluorescence images showing dose-dependent changes in endothelial network formation. Endothelial cells were stained with an anti-CD31 antibody (green), and cell membranes were labeled with a red cell mask; scale bar: 100 μm. ( B ) Quantitative analysis of HUVEC and HPASMC counts, total tube length, and number of junctions following nanoMIL-89 treatment. Nine fields were analyzed per well (n = 6–12 per condition). Data are presented as mean ± SEM. Statistical comparisons were performed using one-way ANOVA; *p < 0.05 vs control.

Article Snippet: HUVECs were maintained in EGM-2 medium (PromoCell, C-22011), and HPASMCs in high-glucose DMEM (Capricorn, DMEM-H-12), both supplemented with 10% fetal bovine serum (FBS; ScienCell, 0500), 1% penicillin/streptomycin (Gibco, 15140122), and 1% endothelial cell growth supplement for HUVECs only (PromoCell, C-39215).

Techniques: Co-Culture Assay, Immunofluorescence, Staining, Labeling, Control

Journal: International Journal of Nanomedicine

Article Title: Bosentan Delivery via Nano Metal-Organic Framework nanoMIL-89 Restores Vascular Homeostasis in Pulmonary Arterial Hypertension

doi: 10.2147/IJN.S535437

Figure Lengend Snippet: Angiogenic effects of free Bosentan at various concentrations in a 3D co-culture model of HUVECs and HPASMCs. ( A ) Representative immunofluorescence images showing endothelial network formation following Bosentan treatment. Endothelial structures were stained with anti-CD31 antibody (green), and cell membranes were labeled with a red cell mask; scale bar: 100 μm. ( B ) Quantitative analysis of HUVEC and HPASMC counts, total tube length, and number of junctions after treatment with increasing doses of Bosentan. Nine fields were quantified per well (n = 6–12 per condition). Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA; *p < 0.05 versus control.

Article Snippet: HUVECs were maintained in EGM-2 medium (PromoCell, C-22011), and HPASMCs in high-glucose DMEM (Capricorn, DMEM-H-12), both supplemented with 10% fetal bovine serum (FBS; ScienCell, 0500), 1% penicillin/streptomycin (Gibco, 15140122), and 1% endothelial cell growth supplement for HUVECs only (PromoCell, C-39215).

Techniques: Co-Culture Assay, Immunofluorescence, Staining, Labeling, Control