lps  (InvivoGen)

Journal: Journal of Translational Autoimmunity

Article Title: Vitamin D3 encapsulated in polymeric nanoparticles to dampen the pro-inflammatory immune response

doi: 10.1016/j.jtauto.2025.100321

Figure Lengend Snippet: VD3-NPs inhibit cytokine secretion in whole blood. (A) Schematic overview of experimental setup. Whole blood from healthy donors was diluted 1:1 and treated with soluble VD3 or VD3-NP for 18 h, after which samples were stimulated with LPS to induce cytokine secretion. After 6 h of LPS stimulation, the supernatant was collected and cytokine concentrations of IL-6, IL-23, IL-10 and TNFα were determined by ELISA. (B) IL-6, IL-23, IL-10 and TNFα levels (pg/ml) in LPS-stimulated samples. Bar graphs depict mean ± SEM and each data point represents a healthy donor (n = 8) from two independent experiments. (∗P > 0.05; ∗∗P > 0.01; ∗∗∗P > 0.001).

Article Snippet: After overnight incubation at 37 °C, lipopolysaccharide (LPS) (100 ng/mL) (vac-3pelps, InvivoGen) or medium was added to the cultures followed by 6 h of incubation at 37 °C.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Autoimmunity

Article Title: Vitamin D3 encapsulated in polymeric nanoparticles to dampen the pro-inflammatory immune response

doi: 10.1016/j.jtauto.2025.100321

Figure Lengend Snippet: VD3-NPs alter IL-8 secretion by neutrophils. (A) Schematic of the experimental setup. Neutrophils were isolated from whole blood from healthy donors and treated with soluble VD3 or VD3-NP in combination with LPS for 2 h to determine degranulation status, or 24 h to determine IL-8 secretion. (B) IL-8 levels (pg/ml) in LPS-stimulated samples (C) CD16, CD62L, CD63, CD66b surface expression levels in LPS-stimulated neutrophils depicted in geometric mean fluorescent intensity (gMFI) normalized to the untreated samples. Bar graphs depict mean ± SEM and each data point represents a healthy donor (n = 9) from two independent experiments. (∗P > 0.05).

Article Snippet: After overnight incubation at 37 °C, lipopolysaccharide (LPS) (100 ng/mL) (vac-3pelps, InvivoGen) or medium was added to the cultures followed by 6 h of incubation at 37 °C.

Techniques: Isolation, Expressing

Journal: Journal of Translational Autoimmunity

Article Title: Vitamin D3 encapsulated in polymeric nanoparticles to dampen the pro-inflammatory immune response

doi: 10.1016/j.jtauto.2025.100321

Figure Lengend Snippet: VD3-NPs attenuate inflammation in monocytes. Monocytes were isolated from buffy coats and treated with soluble VD3 or VD3-NP for 24 h, after which samples were stimulated with LPS for another 24 h. Following incubation, marker expression was analysed using flow cytometry and cytokine levels in the supernatant were determined by ELISA. (A) CD14, CD86, CD11b, and HLA-DR surface expression levels in LPS-stimulated monocytes depicted in bar graphs displaying gMFI. (B) IL-6, IL-23, IL-10 and TNFα levels (pg/ml) in LPS-stimulated samples. Bar graphs depict mean ± SEM and each data point represents a healthy donor (n = 8) from two independent experiments. (∗P > 0.05; ∗∗P > 0.01; ∗∗∗P > 0.001).

Article Snippet: After overnight incubation at 37 °C, lipopolysaccharide (LPS) (100 ng/mL) (vac-3pelps, InvivoGen) or medium was added to the cultures followed by 6 h of incubation at 37 °C.

Techniques: Isolation, Incubation, Marker, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Wound Repair and Regeneration

Article Title: Potential of Heparan Sulphate Mimetics Integrated Into Collagen Scaffolds for Enhanced Skin Wound Healing

doi: 10.1111/wrr.70096

Figure Lengend Snippet: Effect of OTR4120 and/or SHH in collagen scaffolds on primary human macrophages. Type I collagen scaffolds with M0 macrophages were used as control. As secondary controls, Col I with either M1 or M2 macrophages were included. Mean fluorescence intensity (MFI) was measured by flow cytometry for (A) M1 associated markers and (B) M2 associated markers. (C) Cytokine concentrations in the supernatants. Every donor is represented with a different symbol. Data are represented as mean ± SD ( n = 3). p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: As controls, M0 macrophages were stimulated for 24 h with 100 ng/mL lipopolysaccharides (LPS, vac‐3pelps, InvivoGen, San Diego, CA, USA) with 20 ng/mL interferon gamma (IFNγ, PeproTech) to polarize them to M1‐like macrophages or with 20 ng/mL interleukin 4 (IL4, 170–076‐135) + 20 ng/mL interleukin 13 (IL13, 130–112‐412) to polarize them to M2‐like macrophages (both from Miltenyi Biotech).

Techniques: Control, Fluorescence, Flow Cytometry