Journal: Journal of Biomedical Science and Engineering

Article Title: Healing Mechanism and Osteogenic Capacity of Bovine Bone Mineral—Human Amniotic Mesenchymal Stem Celland Autogenous Bone Graft in Critical Size Mandibular Defect

doi: 10.4236/jbise.2015.810070

Figure Lengend Snippet: Figure 7. Result of immunohistochemistry evaluation for thickness of collagen type-I fibers and osteocalcin expression at 12 week. Thickness of collagen type-I fibers was significantly higher in tissue engineering groups compared to ABG group (top left), expression of osteocalcin was significantly higher in BBM-hAMSC group compared to BBM and ABG group (top right). Photos (a), (b) and (c) showing collagen type-I fibers in BBM-hAMSC, BBM and ABG group, respectively (red arrow pointing to collagen type-I fibers, ×400); photos (d), (e) and (f) showing osteocalcin expression in BBM-hAMSC, BBM and ABG group, respectively(red arrow pointing to positive cells, black arrow pointing to negative cells, ×400). note. This finding indicated that VEGF expression in BBM-hAMSC and ABG group might have reached their peaks in the first few days after implantation compared to that in BBM group which started at later stage. This phenomenon was likely to be associated with the presence of living cells inside the scaffold of BBM-hAMSC group and the grafted bone in ABG group. As the BBM scaffold and the non-vascularized bone graft were avascular and hence hypoxic, overexpression of VEGF by the pre-existing cells in those groups was expected to occur in the first few post-implantation days. This presumption was supported by result of the study which demonstrated that exposure of stem cells to hypoxia resulted in up-regulation of VEGF [18]. Further study eva- luating VEGF expression in implanted scaffold earlier than 7 days would be required to prove this presumption. The expression of BMP2 and Runx2 in ABG group which were significantly higher than the tissue engineer- ing groups in the first week after implantation and the decline of those parameters in ABG group as opposed to the increase of the same parameters in tissue engineering groups in the second week (Figure 4 and Figure 5) suggested that osteogenic activities was initiated earlier and stronger in ABG compared to tissue engineering

Article Snippet: Sections were stained with anti VEGF-A antibody (anti-rabbit VEGF polyclonal antibody, USCN, USA; followed by anti-human VEGF monoclonal antibody, LifeSpan BioSciences, Inc. USA), anti BMP2 antibody (anti-rabbit BMP-2 polyclonal antibody, ABIN, USA; followed by anti-human BMP-2 monoclonal antibody, LifeSpan BioSciences, Inc. USA), anti-rabbit Runx-2 monoclonal antibody (Santa Cruz Biotech., USA), anti-rabbit osteocalcin monoclonal antibody (Novus Biological, USA), and anti-rabbit collagen I monoclonal antibody (Novus Biological, USA).

Techniques: Immunohistochemistry, Expressing, Over Expression

Journal: Biomaterials advances

Article Title: 3D-printed porous Ti6Al4V alloys with silver coating combine osteocompatibility and antimicrobial properties.

doi: 10.1016/j.msec.2021.112629

Figure Lengend Snippet: Fig. 7. Human OB expression of osteogenic related genes Runt-related transcription factor 2 (RUNX2), collagen type I alpha 1 (COL1A1) and alpha 2 (COL1A2), alkaline phosphatase (ALP), and osteocalcin (OCN) at 3, 7, 14 and 28 days of cell culture on Porolink (PL), Trabeculink (TL), silver-coated Trabeculink (TLSN) scaffolds, and quantification of mineralization by alizarin red (AR) staining after 14 and 28 days.

Article Snippet: Afterwards, the samples were blocked with normal 10% goat serum (s-1000, Sigma Aldrich, Sweden) solution prepared in PBS containing 2% bovine serum albumin (BSA) and 0.3% Triton X-100 for 30 min. OCN antibody (20 μg/mL mouse anti-human/rat OCN, MAB1419, R&D Systems, United Kingdom) solution in PBS/2%BSA/0.3%Triton X-100was added and incubated overnight at 4 °C.

Techniques: Expressing, Cell Culture, Staining

Journal: Biomaterials advances

Article Title: 3D-printed porous Ti6Al4V alloys with silver coating combine osteocompatibility and antimicrobial properties.

doi: 10.1016/j.msec.2021.112629

Figure Lengend Snippet: Fig. 8. Immunostained images of hOB cultured on Porolink (PL), Trabeculink (TL), silver-coated Trabeculink (TLSN) scaffolds, and control samples consisting of glass coverslips (Ctrl) after 14 and 28 days showing cell nuclei (blue, DAPI), cytosol (green, CFDA), osteocalcin (red, OCN) and merged channels (scale bar: 100 μm).

Article Snippet: Afterwards, the samples were blocked with normal 10% goat serum (s-1000, Sigma Aldrich, Sweden) solution prepared in PBS containing 2% bovine serum albumin (BSA) and 0.3% Triton X-100 for 30 min. OCN antibody (20 μg/mL mouse anti-human/rat OCN, MAB1419, R&D Systems, United Kingdom) solution in PBS/2%BSA/0.3%Triton X-100was added and incubated overnight at 4 °C.

Techniques: Cell Culture, Control

Journal: Breast Cancer Research : BCR

Article Title: Osteoblasts are “educated” by crosstalk with metastatic breast cancer cells in the bone tumor microenvironment

doi: 10.1186/s13058-019-1117-0

Figure Lengend Snippet: EOs are present in patient samples of bone metastatic breast cancer. Human patient samples of bone metastatic breast cancer were stained using multi-plex immunofluorescence for RUNX2 (green), osteocalcin (OCN, red), IL-6 (purple), and alpha-SMA (yellow). Left panel—osteoblast identification: white arrows show osteoblasts positive for both RUNX2 and OCN. Middle panel—“uneducated” and “educated” osteoblast identification: blue arrows show “uneducated” osteoblasts alpha-SMA and IL-6 positive; yellow arrows show “educated” osteoblasts alpha-SMA high, but IL-6 low; purple arrows show “educated” osteoblasts IL-6 high, but alpha-SMA low. Right panel—“educated” osteoblast identification: green arrows show “educated” osteoblasts both IL-6 and alpha-SMA low, DAPI positive. T, tumor; arrows, osteoblast. DAPI, nuclear stain. Scale bar = 50 μm

Article Snippet: The slides were incubated overnight at 4 °C with either mouse anti-human osteocalcin (R&D Systems, 10 μg/ml), rabbit anti-human RUNX2 (Abcam, 1:500), mouse anti-human alpha-smooth muscle actin (1:100, Abcam), rabbit anti-human alkaline phosphatase (1:100, Abcam), rabbit anti-human osteopontin (1:500, Abcam), or goat anti-human IL-6 (40 μg/mL, R&D Systems), Next, slides were incubated for 1 h at room temperature with either donkey anti-mouse 594, chicken anti-rabbit 594, donkey anti-goat 488, or goat anti-rabbit 488 (1:1000, Biotium).

Techniques: Staining, Immunofluorescence

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: International Journal of Molecular Sciences

Article Title: Three-Dimensional Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Promotes Matrix Metallopeptidase 13 (MMP13) Expression in Type I Collagen Hydrogels

doi: 10.3390/ijms222413594

Figure Lengend Snippet: Antibodies used for immunohistochemistry.

Article Snippet: Primary , Mouse monoclonal, anti-OCN , Novus Biologicals, H00000632-M01 , 1:100.

Techniques: Immunohistochemistry

Journal: Journal of Functional Biomaterials

Article Title: Sequential Release of Panax Notoginseng Saponins and Osteopractic Total Flavone from Poly ( L -Lactic Acid) Scaffold for Treating Glucocorticoid-Associated Osteonecrosis of Femoral Head

doi: 10.3390/jfb14010031

Figure Lengend Snippet: Immunohistochemistry staining of OCN after scaffold implantation. ( A ) Distribution and quantity of OCN in decalcified sections (bone tunnel: black circle) at magnifications of 20× and 200×. 200× images are higher magnifications of areas within black boxes. Red arrow indicates OCN positive cells, bars = 500 and 50 μm for 20× and 200×, respectively. ( B ) Quantitative analysis of OCN expression. n = 10 in each group, error bar represents SD. ANOVA test with LSD analysis was used for statistical analysis, * p < 0.05 vs. control group or POP group; ** p < 0.01 vs. control group or POP group.

Article Snippet: After blocking, the sections were incubated with primary antibodies VEGF (1:100; Novus NBP2-45235, Bio-Techne Corporation, Minneapolis, MN, USA), OPN (1:100; Novus NB110-89062, Bio-Techne Corporation, Minneapolis, MN, USA) and OCN (1:100; Novus NBP2-89037, Bio-Techne Corporation, Minneapolis, MN, USA) overnight at 4 °C.

Techniques: Immunohistochemistry, Staining, Expressing, Control