Journal: Frontiers in Immunology

Article Title: In silico design of novel precision vaccine targeting sclerostin epitopes for osteoporosis prevention and treatment

doi: 10.3389/fimmu.2025.1644437

Figure Lengend Snippet: Screening and analysis of high-affinity epitopes on SOST. (A) ELISA experiments were conducted to identify SOST fragments with strong binding affinity for ROMO, revealing that SOST 114–143 and SOST 143–173 exhibit significantly higher affinity ( P <0.01). (B) A schematic diagram delineating the binding functional regions associated with the high-affinity fragments of SOST. (C) ELISA results indicate that SOST 131–163 displays the highest affinity for ROMO ( P <0.01), thereby identifying it as a potent functional epitope of SOST. (D-a) SOST 131–163 fragment (highlighted in yellow) is located within the loop3 domain of SOST protein. (D-b) Docking studies indicate that SOST 131–163 fragment interacts with ROMO light chain, yielding a binding free energy of -25.8 kcal/mol and an interface area of 712.9 Ų. (D-c) Additionally, SOST 131–163 fragment can bind to the ROMO heavy chain, resulting in a binding free energy of -33.19 kcal/mol and an interface area of 451.6 Ų. (E) CTL epitopes within SOST 131–163 sequence include two strong binder epitopes and four weak binder epitopes. (F) HTL epitopes in SOST 131–163 sequence comprise one strong binder epitope and four weak binder epitopes. Predictions of B cell epitopes for SOST 131–163 sequence are illustrated, including predicted linear B cell epitopes (G) and predicted discontinuous B cell epitopes (H) .

Article Snippet: In brief, 1 μg/mL of human SOST protein (MedChemExpress Inc.) was coated onto the wells of MaxiSorp microtiter plates (Thermo Fisher Scientific Inc.) and incubated overnight at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Functional Assay, Sequencing

Journal: Materials Today Bio

Article Title: Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells

doi: 10.1016/j.mtbio.2025.102154

Figure Lengend Snippet: OCy in two quadruple culture experiments under different cultivation conditions. (a, b) Fluorescence microscopic images. Cytoskeleton appears green (iFluor488 phalloidin), nuclei appear blue (DAPI) and DMP1 (a)/SOST (b) appear magenta (Alexa Fluor 546). Scale bars represent 100 μm. (c) Scheme of OCy in quadruple culture. (d) Gene expression of OCy markers presented as ΔCt (each experiment n = 4, in total n = 8). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Gene expression analysis was performed using qPCR reactions with the TaqMan Fast Advanced Master Mix (Applied Biosystems) and TaqMan Gene Expression Assays (all Applied Biosystems) for the following genes: actin-β (ACTB; Hs01060665_g1), alkaline phosphatase (ALPL; Hs01029144_m1), bone gamma-carboxyglutamate protein (osteocalcin/BGLAP; Hs01587814_g1), bone sialoprotein II (BSP II, IBSP; Hs00173720_m1), receptor activator of NF-κB ligand (RANKL/TNFSF11; Hs00243522_m1), osteoprotegerin (OPG/TNFRSF11B; Hs00900358_m1), podoplanin (PDPN; Hs00366766_m1), matrix extracellular phosphoglycoprotein (MEPE; Hs00220237_m1), dentin matrix protein 1 (DMP1; Hs01009391_g1), sclerostin (SOST; Hs00228830_m1), collagen type I alpha 1 chain (COL1A; Hs00164004_m1), bone morphogenetic protein 2 (BMP-2; Hs00154192_m1), platelet endothelial cell adhesion molecule (PECAM1/CD31; Hs01065279_m1), vascular endothelial growth factor (VEGF; Hs00900055_m1), vascular endothelial growth factor receptor 2/kinase insert domain receptor (VEGFR/KDR; Hs00911700_m1), von willebrand factor (vWF; Hs01109446_m1), tartrate-resistant acid phosphatase (ACP5/TRAP; Hs00356261_m1), cathepsin K (CTSK; Hs00166156_m1) and carbonic anhydrase II (CA2; Hs01070108_m1), nuclear factor of activated T-cells cytoplasmic (NFATC1; Hs00542675_m1), dendrocyte expressed seven transmembrane protein (DCSTAMP; Hs00229255_m1), according to manufacturer’s instructions.

Techniques: Fluorescence, Gene Expression

Journal: Materials Today Bio

Article Title: Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells

doi: 10.1016/j.mtbio.2025.102154

Figure Lengend Snippet: OCy in two quadruple culture experiments in comparison to OCy monocultures. (a) Fluorescence microscopic images in quadruple culture. Cytoskeleton appears green (iFluor488 phalloidin), nuclei appear blue (DAPI) and DMP1/SOST/PDPN appear magenta (Alexa Fluor 546). Scale bars represent 100 μm. (b) Gene expression of OCy markers presented as fold change ± upper and lower limit normalized to OCy monocultures (each experiment n = 4). (c) Secreted BGLAP and SOST protein in cell culture supernatants (each experiment n = 3). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. Not detectable (n.d.). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Gene expression analysis was performed using qPCR reactions with the TaqMan Fast Advanced Master Mix (Applied Biosystems) and TaqMan Gene Expression Assays (all Applied Biosystems) for the following genes: actin-β (ACTB; Hs01060665_g1), alkaline phosphatase (ALPL; Hs01029144_m1), bone gamma-carboxyglutamate protein (osteocalcin/BGLAP; Hs01587814_g1), bone sialoprotein II (BSP II, IBSP; Hs00173720_m1), receptor activator of NF-κB ligand (RANKL/TNFSF11; Hs00243522_m1), osteoprotegerin (OPG/TNFRSF11B; Hs00900358_m1), podoplanin (PDPN; Hs00366766_m1), matrix extracellular phosphoglycoprotein (MEPE; Hs00220237_m1), dentin matrix protein 1 (DMP1; Hs01009391_g1), sclerostin (SOST; Hs00228830_m1), collagen type I alpha 1 chain (COL1A; Hs00164004_m1), bone morphogenetic protein 2 (BMP-2; Hs00154192_m1), platelet endothelial cell adhesion molecule (PECAM1/CD31; Hs01065279_m1), vascular endothelial growth factor (VEGF; Hs00900055_m1), vascular endothelial growth factor receptor 2/kinase insert domain receptor (VEGFR/KDR; Hs00911700_m1), von willebrand factor (vWF; Hs01109446_m1), tartrate-resistant acid phosphatase (ACP5/TRAP; Hs00356261_m1), cathepsin K (CTSK; Hs00166156_m1) and carbonic anhydrase II (CA2; Hs01070108_m1), nuclear factor of activated T-cells cytoplasmic (NFATC1; Hs00542675_m1), dendrocyte expressed seven transmembrane protein (DCSTAMP; Hs00229255_m1), according to manufacturer’s instructions.

Techniques: Comparison, Fluorescence, Gene Expression, Cell Culture