Journal: Experimental and Therapeutic Medicine

Article Title: Effect of Porphyromonas gingivalis lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

doi: 10.3892/etm.2021.10087

Figure Lengend Snippet: HPDLCs and HUASMCs were characterized. (A) HPDLCs were positive for anti-vimentin staining. (B) HPDLCs were negative for anti-keratin staining. (C) HUASMCs expressed α-smooth muscle actin. The DAPI staining image is superimposed with green fluorescence, with α-SMA being fluorescent green and the nuclei blue (scale bars, 100 µm). HUASMCs, human umbilical artery smooth muscle cells; HPDLCs, human periodontal ligament cells.

Article Snippet: Primary HUASMCs (cat. no. 8030) were purchased from ScienCell Research Laboratories, Inc. and subcultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ̊C with 5% CO 2 after recovery.

Techniques: Staining, Fluorescence

Journal: Experimental and Therapeutic Medicine

Article Title: Effect of Porphyromonas gingivalis lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

doi: 10.3892/etm.2021.10087

Figure Lengend Snippet: HPDLC-HUASMC co-culture system was established with Transwell inserts. (A) HPDLCs, with micropores in the porous membrane visible. (B) HUASMCs, exhibiting a radial spindle-shaped arrangement when attached to the plate (scale bars, 20 µm). HUASMCs, human umbilical artery smooth muscle cells; HPDLCs, human periodontal ligament cells.

Article Snippet: Primary HUASMCs (cat. no. 8030) were purchased from ScienCell Research Laboratories, Inc. and subcultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ̊C with 5% CO 2 after recovery.

Techniques: Co-Culture Assay, Membrane

Journal: Experimental and Therapeutic Medicine

Article Title: Effect of Porphyromonas gingivalis lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

doi: 10.3892/etm.2021.10087

Figure Lengend Snippet: Cell viability. (A) Effect of Pg-LPS on HUASMC viability in the co-culture system. (B) Cell viability in HUASMCs treated with or without Pg-LPS in the calcified-inducing culture medium. Values are expressed as the mean ± standard deviation of three independent experiments in each group. * P<0.05 vs. HUASMCs (group C); # P<0.05. HUASMCs, human umbilical artery smooth muscle cells; PDLCs, periodontal ligament cells; Pg-LPS, Porphyromonas gingivalis lipopolysaccharide; ALP, alkaline phosphatase.

Article Snippet: Primary HUASMCs (cat. no. 8030) were purchased from ScienCell Research Laboratories, Inc. and subcultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ̊C with 5% CO 2 after recovery.

Techniques: Co-Culture Assay, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Effect of Porphyromonas gingivalis lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

doi: 10.3892/etm.2021.10087

Figure Lengend Snippet: Induction of ALP activity. Effects of the normal medium (A) and the calcification-inducing culture medium (B) on ALP activity of HUASMCs and the HUASMCs-HPDLCs co-culture system. Values are expressed as the mean ± standard deviation of three independent experiments in each group. * P<0.05 vs. HUASMCs in calcification-inducing culture medium (group CC); # P<0.05. HUASMCs, human umbilical artery smooth muscle cells; PDLCs, periodontal ligament cells; Pg-LPS, Porphyromonas gingivalis lipopolysaccharide; ALP, alkaline phosphatase; OD, optical density.

Article Snippet: Primary HUASMCs (cat. no. 8030) were purchased from ScienCell Research Laboratories, Inc. and subcultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ̊C with 5% CO 2 after recovery.

Techniques: Activity Assay, Co-Culture Assay, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Effect of Porphyromonas gingivalis lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

doi: 10.3892/etm.2021.10087

Figure Lengend Snippet: Induction of osteogenic gene transcription in HUASMCs. Transcript levels for the osteogenic genes (A) Runx2, (B) ALP and (C) BSP were determined at 48 h by reverse transcription-quantitative PCR and normalized to the level of GAPDH transcript. Values are expressed as the mean ± standard deviation (n=3). The expression of Runx2, ALP and BSP was significantly higher in co-culture groups treated with Pg-LPS (1 µg/ml) and calcification-inducing medium than in the control groups. * P<0.05 vs. HUASMCs (group C); # P<0.05. HUASMCs, human umbilical artery smooth muscle cells; PDLCs, periodontal ligament cells; Pg-LPS, Porphyromonas gingivalis lipopolysaccharide; ALP, alkaline phosphatase; Runx2, core-binding factor α1; BSP, bone sialoprotein; OD, optical density.

Article Snippet: Primary HUASMCs (cat. no. 8030) were purchased from ScienCell Research Laboratories, Inc. and subcultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ̊C with 5% CO 2 after recovery.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation, Expressing, Co-Culture Assay, Control, Binding Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Effect of Porphyromonas gingivalis lipopolysaccharide on calcification of human umbilical artery smooth muscle cells co-cultured with human periodontal ligament cells

doi: 10.3892/etm.2021.10087

Figure Lengend Snippet: Effect of Pg-LPS on calcified nodule formation of HUASMCs in different groups. (AA) HUASMCs (group C); (AB) HUASMCs + Pg-LPS (group C-P); (AC) PDLCs-HUASMCs (group CO); (AD) PDLCs-HUASMCs + Pg-LPS (group CO-P); (AE) HUASMCs in calcification-inducing culture medium (group C-C); (AF) HUASMCs + Pg-LPS in calcification-inducing culture medium (group C-CP); (AG) PDLCs-HUASMCs in calcification-inducing culture medium (group CO-C); and (AH) PDLCs-HUASMCs + Pg-LPS in calcification-inducing culture medium (group CO-CP). The calcified nodules were scattered in the C-P and CO-P groups and were almost absent in the C and CO groups. All of the groups under calcification induction or calcification induction + Pg-LPS (1 µg/ml) conditions exhibited many calcified nodules, whilst the co-cultured CO-C and CO-CP groups had slightly more calcified nodules than those cultured in DMEM medium (scale bar, 100 μm). The quantified results of calcification based on alizarin red S staining three weeks after calcification induction (B). The calcified nodule formation was significantly higher in calcification-inducing medium and co-culture groups treated with Pg-LPS (1 μg/ml) than in the control groups. Values are expressed as the mean ± standard deviation (n=3). # P<0.05, calcification-inducing culture medium vs. normal culture medium (i.e. group C-C vs. group C; group C-CP vs. group C-P; group CO-C vs. group CO; group CO-CP vs. group CO-P); * P<0.05. HUASMCs, human umbilical artery smooth muscle cells; PDLCs, periodontal ligament cells; Pg-LPS, Porphyromonas gingivalis lipopolysaccharide..

Article Snippet: Primary HUASMCs (cat. no. 8030) were purchased from ScienCell Research Laboratories, Inc. and subcultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ̊C with 5% CO 2 after recovery.

Techniques: Cell Culture, Staining, Co-Culture Assay, Control, Standard Deviation