Journal: Frontiers in Immunology

Article Title: Immune Modulation to Enhance Bone Healing—A New Concept to Induce Bone Using Prostacyclin to Locally Modulate Immunity

doi: 10.3389/fimmu.2019.00713

Figure Lengend Snippet: Immunomodulatory effects of Iloprost on isolated CD8+ T cells. The purity of the isolated CD8+ T cells after bead-based separation as well as after in vitro stimulation was confirmed by flow cytometry (A) . CD8+ T cells were stimulated by α-CD3/α-CD28 in addition to either PBS or 3 μM Iloprost. The following secreted cytokine concentrations were evaluated: TNFα (B) and IFNγ (C) ; n = 6.

Article Snippet: The isolation was performed via the CD8 S-pluriBeads anti-ms kit (pluriSelect Life Sciences, Leipzig, Germany).

Techniques: Isolation, In Vitro, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Immune Modulation to Enhance Bone Healing—A New Concept to Induce Bone Using Prostacyclin to Locally Modulate Immunity

doi: 10.3389/fimmu.2019.00713

Figure Lengend Snippet: Osteoimmunological effect of Iloprost treated isolated CD8+ T cells. Isolated α-CD3/α-CD28 activate CD8+ T cells were cultured either with Iloprost or PBS as control. Obtained conditioned media was added to an osteogenic differentiation culture of MSCs and the secretion of mineralized matrix was measured by Alizarin Red staining. (A) Representative images of the Alizarin Red staining of the different culture conditions. (B) Quantification of the Alizarin Red signal. OM, osteoinductive media. Scale bars: 200 μm; n = 6.

Article Snippet: The isolation was performed via the CD8 S-pluriBeads anti-ms kit (pluriSelect Life Sciences, Leipzig, Germany).

Techniques: Isolation, Cell Culture, Control, Staining

Journal: Frontiers in Immunology

Article Title: Immune Modulation to Enhance Bone Healing—A New Concept to Induce Bone Using Prostacyclin to Locally Modulate Immunity

doi: 10.3389/fimmu.2019.00713

Figure Lengend Snippet: Immunohistochemical (IHC) analysis of the distribution of immune cells in the fracture zone 3 days post-osteotomy. (A) A section of a fractured femur from the Iloprost treated group is displayed, Movat Pentachrome staining. The region of interest (ROI) for the quantitative analysis of IHC images is enlarged (red rectangle). The black rectangle represents the region from which the representative examples of the respective IHC staining for (B) CD4+ and CD8+ T cells and osteoblasts, (C) IFNγ producing CD8+ T cells; CD8+IFNγ+ cells = white indicator, (D) macrophages; M1-type macrophages = white indicator; M2-type macrophages = yellow indicator, and (E) osteoclasts were taken. The quantification of the cellular distributions are displayed in the corresponding dot plot graphs. Color code: Movat Pentachrome mineralized bone = yellow, cartilage = dark blue-green, connective tissue = light blue-green, muscle fibers = light orange, cell nuclei = purple, fibrin clot = red/orange; (B) osteoblasts = white, CD8+ T cells = red, CD4+ T cells = green and cell nuclei = blue; (C) osteoblasts = white, CD8+ T cells = red, IFNγ = green, cell nuclei = blue; (D) CD68 = green, CD80 = white, CD206 = red; (E) osteoclasts: double positive for cathepsinK = white and CD68 = green cell nuclei = blue. Scale bars: 20 μm (IHC) 1,000 μm (Movat Pentachrome); n = 6.

Article Snippet: The isolation was performed via the CD8 S-pluriBeads anti-ms kit (pluriSelect Life Sciences, Leipzig, Germany).

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry