Journal: International Journal of Oral Science

Article Title: circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

doi: 10.1038/s41368-022-00184-2

Figure Lengend Snippet: circ_0003204 negatively regulated hASC osteogenesis. a Images of ALP and ARS stainings after transfection (scale bar = 200 μm). b – d Western blot after 7-day osteogenic induction of hASCs detected the protein expressions and the results were quantitated. e , f The expressions of ALPL and RUNX2 after 7-day osteogenic induction of hASCs were evaluated by RT-qPCR. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.000 1)

Article Snippet: After 14-day osteogenic differentiation, the mineralized nodules were measured by ARS staining using 0.1% ARS staining solution (pH = 4.2, Cyagen).

Techniques: Transfection, Western Blot, Quantitative RT-PCR

Journal: International Journal of Oral Science

Article Title: circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

doi: 10.1038/s41368-022-00184-2

Figure Lengend Snippet: circ_0003204 inhibited hASC osteogenesis through sponging miR-370-3p. a Bioinformatic analysis predicted the target relationship. b RT-qPCR evaluated the expression of miR-370-3p. c The putative binding sequences. d Luciferase reporter genes were co-transfected with miR‐370‐3p mimic/mimic-NC and mut-type/wild-type circ_0003204 into cells, and the luciferase activities were measured. e RT-PCR evaluated the expression of circ_0003204 and miR-370-3p. f Images of ALP and ARS stainings after transfection. g , h ALP and ARS activity analyses were measured. i RT-qPCR evaluated the mRNA expressions after 7-day osteogenic induction of hASCs (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.000 1)

Article Snippet: After 14-day osteogenic differentiation, the mineralized nodules were measured by ARS staining using 0.1% ARS staining solution (pH = 4.2, Cyagen).

Techniques: Quantitative RT-PCR, Expressing, Binding Assay, Luciferase, Transfection, Reverse Transcription Polymerase Chain Reaction, Activity Assay

Journal: International Journal of Oral Science

Article Title: circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

doi: 10.1038/s41368-022-00184-2

Figure Lengend Snippet: miR-370-3p regulated hASC osteogenesis by targeting HDAC4. a Bioinformatic analysis predicted the target relationship. b The expressions of HDAC4 were evaluated by RT-qPCR. c circ_0003204 siRNAs carried Cy3 reporter (red) and transfection efficiency was confirmed by fluorescence microscopy (scale bar = 100 μm). d Images of ALP and ARS stainings after transfection. e , f ALP and ARS activity analyses were detected. g RT-qPCR evaluated the mRNA expression levels of BGLAP after 7-day osteogenic induction of hASCs. h The expression of HDAC4 was regulated by circ_0003204. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.000 1)

Article Snippet: After 14-day osteogenic differentiation, the mineralized nodules were measured by ARS staining using 0.1% ARS staining solution (pH = 4.2, Cyagen).

Techniques: Quantitative RT-PCR, Transfection, Fluorescence, Microscopy, Activity Assay, Expressing

Journal: International Journal of Oral Science

Article Title: circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

doi: 10.1038/s41368-022-00184-2

Figure Lengend Snippet: HDAC4 inhibited hASC osteogenesis. a , b Images of ALP and ARS stainings after transfection (scale bar = 200 μm). c Western blot evaluated the protein expressions after 7-day osteogenic induction of hASCs and histograms show quantification of the band intensities. d RT-qPCR evaluated the mRNA expressions after 7-day osteogenic induction of hASCs. e , f Immunofluorescence (IF) staining of COL1A1 (green) and RUNX2 (red) was performed after 7-day osteogenic differentiation (scale bar = 40 μm). The 4’,6-diamidino-2-phenylindole (DAPI) stained nuclei (blue) (* P < 0.05, ** P < 0.01, **** P < 0.000 1)

Article Snippet: After 14-day osteogenic differentiation, the mineralized nodules were measured by ARS staining using 0.1% ARS staining solution (pH = 4.2, Cyagen).

Techniques: Transfection, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: International Journal of Oral Science

Article Title: circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

doi: 10.1038/s41368-022-00184-2

Figure Lengend Snippet: Characterization of GelMA and coculture of hASCs in GelMA. a The diameter distribution of GelMA. b , c The SEM image exhibited the three-dimensional structure of GelMA and the morphology of hASCs on the surface of GelMA. d Confocal microscopy was used to observe the morphology of hASCs encapsulated in the GelMA (scale bar = 100 μm). e Images of Calcein-AM/PI staining, where calcein-AM indicated live cells with green fluorescence and PI indicated dead cells with red fluorescence (scale bar = 200 μm). f Proliferation ability of hASCs encapsulated in GelMA scaffold was measured using CCK-8. g Images of ALP and ARS stainings after transfection

Article Snippet: After 14-day osteogenic differentiation, the mineralized nodules were measured by ARS staining using 0.1% ARS staining solution (pH = 4.2, Cyagen).

Techniques: Confocal Microscopy, Staining, Fluorescence, CCK-8 Assay, Transfection

Journal: Frontiers in Cell and Developmental Biology

Article Title: Curcumin-primed periodontal ligament stem cells-derived extracellular vesicles improve osteogenic ability through the Wnt/β-catenin pathway

doi: 10.3389/fcell.2023.1225449

Figure Lengend Snippet: Effect of Cur-PDLSC-EV and PDLSC-EV on osteogenic differentiation. ARS and AKP staining in culture dishes (A) and under an inverted microscope (B) . (C) Result of ARS and AKP semi-quantitative analysis after osteogenic induction. (D) Relative mRNA levels of osteogenesis marker genes (COL1, Runx2, and ALP) detected using RT-qPCR.

Article Snippet: Subsequently, the ARS staining solution (Beyotime, China) and AKP staining solution (Beyotime, China) were added and stained for 5 and 30 min, respectively.

Techniques: Staining, Inverted Microscopy, Marker, Quantitative RT-PCR

Journal: RSC Advances

Article Title: Polydopamine-functionalized acellular fish scale scaffolds for accelerated bone tissue regeneration †

doi: 10.1039/d5ra01932j

Figure Lengend Snippet: Osteogenic differentiation ability of scaffolds in vitro . (A) ALP staining and ALP activity of BMSCs after 7 and 14 days of osteogenic induction, and ARS staining and quantitative analysis of calcium nodules after 21 days of osteogenic induction. Scale bar = 200 μm. (B and C) Fluorescence microscopy images and quantitative analysis of IF staining of RUNX2, OPN, and COL-1α in BMSCs after 14 days of osteogenic induction. Scale bar = 100 μm. (D) The mRNA expression levels of RUNX2, OPN, and COL-1α in BMSCs after 14 days of osteogenic induction. All values are presented as means ± standard deviation (ns: no significant differences, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001).

Article Snippet: After 21 days of osteogenic induction, cells were fixed with 4% paraformaldehyde and stained using an ARS kit (Solarbio, China) ( n = 3).

Techniques: In Vitro, Staining, Activity Assay, Fluorescence, Microscopy, Expressing, Standard Deviation

Journal: Heliyon

Article Title: Bone marrow fatty acids affect osteoblastic differentiation through miR-92b-3p in the early stages of postmenopausal osteoporosis

doi: 10.1016/j.heliyon.2023.e16513

Figure Lengend Snippet: The Effects of fatty acids on osteogenic differentiation of MC3T3-E1 cells (A) ARS and ALP staining after culture for 14 days. (B) Relative mRNA expression level of osteoblastic-related genes after culture for 14 days. MTT assay of palmitoleate (C), pentadecanoate (D) and palmitate (E). Relative mRNA expression level of osteoblastic-related genes after treatment with palmitoleate (F), pentadecanoate (G) and palmitate (H). *p < 0.05, **p < 0.01, ***p < 0.001. Experiments were replicated three times.

Article Snippet: After washing three times with double-distilled H 2 O, the cells were incubated with ARS staining solution (Shanghai Yuanye Biotechnology Co., Ltd., Shanghai, China) for 30 min or stained with the BCIP/NBT Alkaline Phosphatase Color Development Kit (Beyotime Institute of Biotechnology) at room temperature according to the manufacturer's instructions.

Techniques: Staining, Expressing, MTT Assay

Journal: Heliyon

Article Title: Bone marrow fatty acids affect osteoblastic differentiation through miR-92b-3p in the early stages of postmenopausal osteoporosis

doi: 10.1016/j.heliyon.2023.e16513

Figure Lengend Snippet: MiR-92b-3p regulates the osteogenic differentiation of MC3T3-E1 cells by targeting PTEN. (A) The expression of miR-92b-3p during osteogenesis. (B) Relative expression level of osteoblastic-related genes after transfection with miR-92b-3p mimic or inhibitor. (C) ARS and ALP staining after transfection with miR-92b-3p mimic or inhibitor. (D) The binding sites between miR-92b-3p and PTEN. (E) The mRNA expression of PTEN after transfection with miR-92b-3p mimic or inhibitor. (F) Luciferase reporter assay exploring the relationship between miR-92b-3p and PTEN. *p < 0.05, **p < 0.01, ***p < 0.001. Experiments were replicated three times.

Article Snippet: After washing three times with double-distilled H 2 O, the cells were incubated with ARS staining solution (Shanghai Yuanye Biotechnology Co., Ltd., Shanghai, China) for 30 min or stained with the BCIP/NBT Alkaline Phosphatase Color Development Kit (Beyotime Institute of Biotechnology) at room temperature according to the manufacturer's instructions.

Techniques: Expressing, Transfection, Staining, Binding Assay, Luciferase, Reporter Assay

Journal: Heliyon

Article Title: Bone marrow fatty acids affect osteoblastic differentiation through miR-92b-3p in the early stages of postmenopausal osteoporosis

doi: 10.1016/j.heliyon.2023.e16513

Figure Lengend Snippet: Fatty acids affected the osteoblastogenesis of MC3T3-E1 cells through miR-92b-3p. (A) The expression of miR-92b-3p after treatment with fatty acids. Relative expression level of osteoblastic-related genes after transfection with miR-92b-3p mimic or inhibitor in the palmitate (B), pentadecanoate (C) and palmitoleate (D) groups. (E) ARS and ALP staining after transfection with miR-92b-3p mimic or inhibitor in the palmitate, pentadecanoate and palmitoleate groups. *p < 0.05, **p < 0.01, ***p < 0.001. Experiments were replicated three times.

Article Snippet: After washing three times with double-distilled H 2 O, the cells were incubated with ARS staining solution (Shanghai Yuanye Biotechnology Co., Ltd., Shanghai, China) for 30 min or stained with the BCIP/NBT Alkaline Phosphatase Color Development Kit (Beyotime Institute of Biotechnology) at room temperature according to the manufacturer's instructions.

Techniques: Expressing, Transfection, Staining