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: PLoS ONE

Article Title: Synergistic Actions of Hematopoietic and Mesenchymal Stem/Progenitor Cells in Vascularizing Bioengineered Tissues

doi: 10.1371/journal.pone.0003922

Figure Lengend Snippet: A. Hematopoietic stem/progenitor cells (HSCs) propagated in suspension culture, assuming spherical shape. B. HSCs form endothelial-like colonies in fibronectin-coated plates. C. Formation of tubular intercellular structures in 3D Matrigel culture. Uptake of acetylated-LDLs (red) (blue: DAPI) (D) and vWF (von Willebrand Factor) immunofluorescent stain (green) (E). F. Quantification of vWF measured by ELISA showing substantial expression of vWF in HSC-derived endothelial-like cells, in comparison with dermal fibroblasts as controls.

Article Snippet: Osteocalcin was detected using a human osteocalcin ELISA kit (R&D Systems, Minneapolis, MN) and von Willebrand factor (vWF) using human vWF-specific ELISA (Helena Laboratories, Beaumont, TX).

Techniques: Suspension, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Derivative Assay, Comparison

Journal: PLoS ONE

Article Title: Synergistic Actions of Hematopoietic and Mesenchymal Stem/Progenitor Cells in Vascularizing Bioengineered Tissues

doi: 10.1371/journal.pone.0003922

Figure Lengend Snippet: Human osteocalcin immunostaining displays increased levels of expression in MSC and HSC co-transplantation sample (C) and VEGF-delivered MSC and HSC co-transplantation sample (D), in contrast with MSC transplantation alone sample (A) and VEGF-delivered MSC transplantation sample (B). Osteoblast-like cells are observed lining the calcium phosphate scaffold (CP) (black arrow in D). E. Quantified human osteocalcin content confirms significantly increased human osteocalcin expression of the co-transplantation group of MSCs and HSCs (n = 5, p<0.05). Interestingly, VEGF delivery decreased human osteocalcin expression, despite the higher level of vascularization (see detailed discussion in text). F–I. Undecalcified H&E stained sections of in vivo implanted scaffolds show increased dark mineralized tissue throughout the co-transplanted MSC and HSCs cell-seeded scaffold (yellow arrows in H) and VEGF-delivered, co-transplanted MSC and HSC-seeded scaffold (yellow arrows in I) groups, in contrast to MSC transplantation alone (F) and VEGF-only (G).

Article Snippet: Osteocalcin was detected using a human osteocalcin ELISA kit (R&D Systems, Minneapolis, MN) and von Willebrand factor (vWF) using human vWF-specific ELISA (Helena Laboratories, Beaumont, TX).

Techniques: Immunostaining, Expressing, Transplantation Assay, Staining, In Vivo

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: International journal of molecular sciences

Article Title: Methodology and Characterization of a 3D Bone Organoid Model Derived from Murine Cells.

doi: 10.3390/ijms25084225

Figure Lengend Snippet: Figure 2. Relative expression screening of key osteoblast and osteoclast markers by in response to differentiation media. (A) RUNX2. (B) TRAP. Relative fluorescence intensity associated with protein expression evaluated from MC3T3-E1 and RAW 264.7 which were grown in 2D culture with exposure to media containing mixtures of osteoblast (BMP2) or osteoclast (RANKL/M-CSF) differentiating factors as compared to complete culture media (negative).. Fluorescence intensity for each condition was assessed by flow cytometry. Raw fluorescence data were normalized to a cell line specific unstained control for each group. Statistical significance was evaluated by one-way ANOVA (A,B) or unpaired T test (** = p < 0.01; **** = p < 0.0001). (C) Qualitative assessment of relative gene expression (2−∆∆Ct) of osteoblastogenic (ALPL, Runx2, Sp7, Bglap, Col1a1) and osteoclastogenic (Trap) genes of undifferentiated cell lines (pooled) versus cells differentiated (pooled) in two-dimensional culture for (MC3T3-E1: day 14; RAW 264.7: day 5). Where appropriate, data are summarized as the sample mean, and error bars represent the sample standard deviation.

Article Snippet: Osteocalcin/Bglap (Bglap) ELISA assays (Novus Biologicals, Catalogue# NBP2-68151, Centennial, CO, USA) were performed per the vendor-supplied protocol.

Techniques: Expressing, Fluorescence, Flow Cytometry, Control, Gene Expression, Standard Deviation