Journal: International Journal of Stem Cells

Article Title: Cannabidiol Promotes Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells in the Inflammatory Microenvironment via the CB2-dependent p38 MAPK Signaling Pathway

doi: 10.15283/ijsc21152

Figure Lengend Snippet: CBD promotes osteogenic differentiation of BMSCs in the inflammatory microenvironment. (A) BMSCs were co-treated with LPS (10 μg/ml) and different concentrations of CBD (0.5, 2.5, 5 μM), as indicated for 1, 3, 5, and 7 days, then ALP activity was detected (n=5). (B) BMSCs were co-treated with LPS and CBD (0.5, 2.5, 5 μM), as indicated for 7 days. Western blot was performed for detecting Runx2, ALP, and OCN. β-actin was used as the internal control (n=3). (C) Quantitative analysis of (B). (D) BMSCs were co-treated with LPS and CBD (0.5, 2.5, 5 μM), as indicated for 7 days. Then, BMSCs were harvested for detecting the mRNA expression levels of Runx2, ALP, and OCN by qRT-PCR. β-actin was used as the internal control (n=5). CBD: cannabidiol, BMSCs: bone mesenchymal stem cells, Runx2: runt-related transcription factor 2, ALP: alkaline phosphatase, OCN: osteocalcin, qRT-PCR: quantitative real-time polymerase chain reaction. Data are represented as means±SD. *p<0.05 compared with the LPS group; # p<0.05 compared with the LPS plus 0.5 μM CBD group.

Article Snippet: The primary antibodies used were: cannabinoid receptor 1 (CB1; DF4918, dilution: 1:1,000, Affinity), cannabinoid receptor 2 (CB2; DF8646, dilution: 1:1,000, Affinity), phosphorylated p38 (p-p38; #4511, dilution: 1:1,000, Cell Signaling Technology), p38 (#8690, dilution: 1:1,000, Cell Signaling Technology), Runt-related transcription factor 2 (Runx2; AF5186, dilution: 1:1,000, Affinity), ALP (DF12525, dilution: 1:1,000, Affinity), osteocalcin (OCN; DF12303, dilution: 1:1,000, Affinity) and β-actin (BM0627, dilution: 1:200, Boster, China).

Techniques: Activity Assay, Western Blot, Control, Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Role of glucocorticoid-induced leucine zipper (GILZ) in inflammatory bone loss

doi: 10.1371/journal.pone.0181133

Figure Lengend Snippet: ( A ) Representative images of H&E stained femur of TNF and TNF-GILZ Tg mice (enlarged boxed areas are shown on right). ( B ) Representative IHC staining images of femurs. Boxed areas are enlarged (right) and arrows indicate osteocalcin-positive osteoblast cells. GP: growth plate; E: endocortical bone. ( C ) Bar graph showing quantified results of A. A Bioquant osteo image analysis system (Bioquant, Nashville, TN) was used to count the number of osteoblasts and measure the oasteoblast covered area. A defined region of interest was established ~0.5mm proximal to the distal growth plate and extended a further 0.5mm, all within the endocortical edges at 50x magnification. A total of 4 to 5 samples were analyzed for each group. ( D ) Representative histological images of paw-joint of TNF and TNF-GILZ Tg mice (20x). C: Cartilage damage; R: Bone resorption; I: inflammation; O: Osteophyte. Scale bar = 100μm.

Article Snippet: The sections were then incubated with an anti-osteocalcin polyclonal antibody (PB9919, Boster Bio, Pleasanton, CA) diluted at 1:1000 in 2% normal goat serum overnight at 4°C.

Techniques: Staining, Immunohistochemistry

Journal: Journal of Orthopaedic Surgery and Research

Article Title: T. gondii excretory proteins promote the osteogenic differentiation of human bone mesenchymal stem cells via the BMP/Smad signaling pathway

doi: 10.1186/s13018-024-04839-0

Figure Lengend Snippet: TgEP promoted the osteogenic differentiation of hBMSCs. ( A ) Cytotoxic effect of TgEP on hBMSCs detected using a cytotoxicity detection kit. ( B ) Effect of TgEP on cell proliferation assessed using a CCK-8 kit. ( C ) ALP staining was performed to evaluate ALP expression in the cells. Scale bar = 500 μm. ( D ) Quantification of ALP staining results using ImageJ software and statistical analysis. ( E ) Alizarin red staining was performed to measure the content of mineralized nodules in the cells. Scale bar = 500 μm. ( F ) Quantification of alizarin red staining results using ImageJ software and statistical analysis. ( G ) mRNA transcription levels of osteogenic genes were assessed using qRT‒PCR after the cells were treated for 3 or 7 days. ( H , J ) Western blotting analysis of the osteogenic proteins ALP, Runx2, Osx, and OCN in hBMSCs treated for 5 days ( H ) or 7 days ( J ). The expression levels were quantified using β-actin as a loading control ( I , K ). ( L ) Immunofluorescence staining was performed to detect the expression of Runx2 in the cells. Scale bar = 30 μm. (Data are presented as the mean ± SD; n = 3; * P < 0.05, ** P < 0.01, *** P < 0.001, ns P >0.05 compared to the control group)

Article Snippet: For immunohistochemistry analysis, tissue sections were stained according to standard protocols using anti-osteocalcin (OCN) antibodies (Cat.#: 614,487) (1:100, ZEN-BIO, China).

Techniques: CCK-8 Assay, Staining, Expressing, Software, Western Blot, Control, Immunofluorescence

Journal: Journal of Orthopaedic Surgery and Research

Article Title: T. gondii excretory proteins promote the osteogenic differentiation of human bone mesenchymal stem cells via the BMP/Smad signaling pathway

doi: 10.1186/s13018-024-04839-0

Figure Lengend Snippet: TgEP promotes the repair of bone defects in vivo. ( A ) Histological analysis of bone tissue sections at 6 weeks, including H&E staining, Masson’s trichrome staining, and osteocalcin immunohistochemical staining. Scale bar = 500 μm. ( B ) Histological analysis of bone tissue sections at 8 weeks, including H&E staining, Masson’s trichrome staining, and osteocalcin immunohistochemical staining. Scale bar = 500 μm.

Article Snippet: For immunohistochemistry analysis, tissue sections were stained according to standard protocols using anti-osteocalcin (OCN) antibodies (Cat.#: 614,487) (1:100, ZEN-BIO, China).

Techniques: In Vivo, Staining, Immunohistochemical staining