Journal: Advanced Healthcare Materials

Article Title: Ascorbic Acid Modulates Collagen Properties in Glucocorticoid‐Induced Osteoporotic Bone: Insights into Chemical, Mechanical, and Biological Regulation

doi: 10.1002/adhm.202502606

Figure Lengend Snippet: AA enhances COL1A1 biological activity and restores osteoblast function and COL1A1 regulation in the presence of Psl. A) ALP staining (red) of osteo‐spheroids embedded in COL1A1 matrices, cultured for 7 d with (+AA), and in the Psl presence (+Psl) or absence (–Psl); Scale bar: 300 µm. B) Quantification of Mean Intensity of ALP (A.U.). C) Alzarin Red (AR) activity assay of osteo‐spheroids after 7 d with or without additional AA. Scale bar, 400 µm. D) Graph showing quantification of Mean Intensity of AR (A.U.). E) OsteoImaging mineralization assay quantifications of Mean Intensity (A.U.) of osteo‐spheroids incubated with or without additional AA for 7 d. F) Gene expression analysis of osteogenic markers ( BGLAP, DMP1, DLX3, RUNX2 ) of osteo‐spheroids embedded in COL1A1 matrices, cultured for 7 d with (+AA), and in the Psl presence (+Psl) or absence (–Psl); Heatmap demonstrates the ∆CT averages. G) Gene expression analysis of COL1A1‐related genes ( PLOD1, PLOD3, DLX3, P3H1, P3H2, P3H3, LOX, SVCT2, COL1A2, COL22A1, IBSP, P4HA2, P4HA3, IFITM5 ) in osteo‐spheroids embedded in COL1A1 matrices, cultured for 7 d with or without ascorbic acid (AA), and in the presence (+Psl) or absence (–Psl) of prednisolone (Psl); Heatmap demonstrates the ∆CT averages H) Fold change of top collagen‐related putative metabolites elevated in the presence of endogenous ascorbic acid (+AA) within the 3D matrix of osteo‐spheroids after 7 d of Psl (+Psl) treatment, based on untargeted metabolomics analysis; fold change threshold > 2 with significance; FDR p‐value < 0.05. The data are expressed as mean ± SD; N = 3, n = 3; significant differences: * * p‐value < 0.01; *** p‐value < 0.005 ; **** p‐value < 0.0001.

Article Snippet: Human osteoblasts (HOBs) (PromoCell) were cultured in osteoblast growth medium (GM) (PromoCell) and differentiated in osteoblast mineralization medium (MM) (PromoCell).

Techniques: Activity Assay, Staining, Cell Culture, Mineralization Assay, Incubation, Gene Expression

Journal: Pancreas

Article Title: Stromal Fibrosis and Expression of Matricellular Proteins Correlate With Histological Grade of Intraductal Papillary Mucinous Neoplasm of the Pancreas

doi: 10.1097/mpa.0000000000000617

Figure Lengend Snippet: FIGURE 2. Area of positive staining revealed by immunohistochemistry in IPMN. Positive staining of the periductal stroma with anti–α-SMA (A), antiperiostin (B), and antigalectin-1 (C) at a magnification of 200. Positive staining of the acinar area with anti–α-SMA (D), antiperiostin (E), and antigalectin-1 (F) at a magnification of 100.

Article Snippet: The primary antibodies and dilutions employed were the following: anti–α-SMA (M0851; DAKO Japan, Tokyo, Japan, 1:50), antiperiostin (RD181045050; BioVendor, Czech Republic, 1:4000), and antigalectin-1 (sc-166618; Santa Cruz Biotechnology, Dallas, 1:40).

Techniques: Staining, Immunohistochemistry

Journal: Advanced Healthcare Materials

Article Title: Ascorbic Acid Modulates Collagen Properties in Glucocorticoid‐Induced Osteoporotic Bone: Insights into Chemical, Mechanical, and Biological Regulation

doi: 10.1002/adhm.202502606

Figure Lengend Snippet: Glucocorticoids impair osteoblast function and collagen expression in vivo. A) H&E staining of mouse bone after 60 days without (Veh) or with the GC, prednisolone (Psl); Scale bar: 1000 µm. B) Representative images for Alkaline phosphatase (ALP) staining of mouse bones; Scale bar, 400 µm; Magnification: Scale bar, 100 µm. C) Quantitative analysis of the ALP‐positive surface intensity in bone tissue sections for n = 5 mice. D) Representative images for Verhoeff van Geison staining showing general elastin‐ (black) and collagen‐specific proteins (red) of mouse bones; Scale bar, 400 µm; Magnification: Scale bar: 100 µm. E) Semi‐quantitative analysis of the collagen fibers‐positive surface intensity in bone tissue sections; n = 5 mice. F) Representative image of collagen type I alpha I (COL1A1) staining of bone sections. Yellow arrows indicate COL1A1 protein expression (red). Green diamonds indicate the interior of cortical bone, near the bone marrow. Scale bar, 50 µm. G) Graph showing the COL1A1 protein expression levels (artificial units; A.U.) in control and Psl‐treated bone tissues. H) Metabolomics KEGG visualization of in vitro osteo‐spheroids treated without (Veh) and with Psl at 100 µm for 7 d. I,J) Analysis of collagen‐related putative metabolites after significance analysis using Metaboanalyst only reported fold‐changes with p‐value < 0.05. Data are expressed as mean ± SD. N = 5, n = 3; significant differences: * p‐value < 0.05.

Article Snippet: Human osteoblasts (HOBs) (PromoCell) were cultured in osteoblast growth medium (GM) (PromoCell) and differentiated in osteoblast mineralization medium (MM) (PromoCell).

Techniques: Expressing, In Vivo, Staining, Control, In Vitro

Journal: Advanced Healthcare Materials

Article Title: Ascorbic Acid Modulates Collagen Properties in Glucocorticoid‐Induced Osteoporotic Bone: Insights into Chemical, Mechanical, and Biological Regulation

doi: 10.1002/adhm.202502606

Figure Lengend Snippet: AA enhances COL1A1 biological activity and restores osteoblast function and COL1A1 regulation in the presence of Psl. A) ALP staining (red) of osteo‐spheroids embedded in COL1A1 matrices, cultured for 7 d with (+AA), and in the Psl presence (+Psl) or absence (–Psl); Scale bar: 300 µm. B) Quantification of Mean Intensity of ALP (A.U.). C) Alzarin Red (AR) activity assay of osteo‐spheroids after 7 d with or without additional AA. Scale bar, 400 µm. D) Graph showing quantification of Mean Intensity of AR (A.U.). E) OsteoImaging mineralization assay quantifications of Mean Intensity (A.U.) of osteo‐spheroids incubated with or without additional AA for 7 d. F) Gene expression analysis of osteogenic markers ( BGLAP, DMP1, DLX3, RUNX2 ) of osteo‐spheroids embedded in COL1A1 matrices, cultured for 7 d with (+AA), and in the Psl presence (+Psl) or absence (–Psl); Heatmap demonstrates the ∆CT averages. G) Gene expression analysis of COL1A1‐related genes ( PLOD1, PLOD3, DLX3, P3H1, P3H2, P3H3, LOX, SVCT2, COL1A2, COL22A1, IBSP, P4HA2, P4HA3, IFITM5 ) in osteo‐spheroids embedded in COL1A1 matrices, cultured for 7 d with or without ascorbic acid (AA), and in the presence (+Psl) or absence (–Psl) of prednisolone (Psl); Heatmap demonstrates the ∆CT averages H) Fold change of top collagen‐related putative metabolites elevated in the presence of endogenous ascorbic acid (+AA) within the 3D matrix of osteo‐spheroids after 7 d of Psl (+Psl) treatment, based on untargeted metabolomics analysis; fold change threshold > 2 with significance; FDR p‐value < 0.05. The data are expressed as mean ± SD; N = 3, n = 3; significant differences: * * p‐value < 0.01; *** p‐value < 0.005 ; **** p‐value < 0.0001.

Article Snippet: Human osteoblasts (HOBs) (PromoCell) were cultured in osteoblast growth medium (GM) (PromoCell) and differentiated in osteoblast mineralization medium (MM) (PromoCell).

Techniques: Activity Assay, Staining, Cell Culture, Mineralization Assay, Incubation, Gene Expression

Journal: BMC Veterinary Research

Article Title: Comparative analysis of the surface exposed proteome of two canine osteosarcoma cell lines and normal canine osteoblasts

doi: 10.1186/1746-6148-9-116

Figure Lengend Snippet: Quantitative real-time PCR of cell lines. Quantitative real-time PCR indicating relative expression of selected genes corresponding to surface proteins detected by mass spectrometry of cultured normal canine osteoblasts (CnOb) and two validated canine osteosarcoma cell lines (POS and HMPOS).

Article Snippet: The canine osteosarcoma cell lines POS and HMPOS [ ] were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and the normal canine osteoblast cell line, CnOb (Cell Applications, San Diego, CA), was cultured in canine osteoblast medium (Cell Applications, San Diego, CA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Mass Spectrometry, Cell Culture

Journal: BMC Veterinary Research

Article Title: Comparative analysis of the surface exposed proteome of two canine osteosarcoma cell lines and normal canine osteoblasts

doi: 10.1186/1746-6148-9-116

Figure Lengend Snippet: Western blot of cell lines. Western blot of whole cell lysate to detect the presence of CD44, Thrombospondin-1 and CYR61 in cultured normal canine osteoblasts (CnOb) and two validated canine osteosarcoma cell lines (POS and HMPOS).

Article Snippet: The canine osteosarcoma cell lines POS and HMPOS [ ] were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and the normal canine osteoblast cell line, CnOb (Cell Applications, San Diego, CA), was cultured in canine osteoblast medium (Cell Applications, San Diego, CA).

Techniques: Western Blot, Cell Culture