Journal: PLOS ONE

Article Title: Evaluation of decellularization process for developing osteogenic bovine cancellous bone scaffolds in-vitro

doi: 10.1371/journal.pone.0283922

Figure Lengend Snippet: Bar charts indicate comparison between DCC scaffold vs DCC scaffold + cells and DMB scaffolds vs DMB scaffold + cells by direct method (a). Bar charts indicate comparison between DMB conditioned media (CM) and DCC conditioned media (CM) vs complete culture media (Control) by indirect method (b). *p<0.05 is indicated. n = 4; CM: conditioned media. (c). HOB cells seeded on DMB and DCC scaffolds showing optimum cell attachment by DAPI staining. (i & iii) DMB scaffolds and (ii & iv) DCC scaffolds; images were captured by fluorescence and confocal microscopy, respectively. Scanning electron micrograph of HOB cells seeded onto DMB (d) and DCC (e) scaffolds. The osteoblast shows a healthy morphology residing on the ECM with evidence of filopodia spread (red arrows).

Article Snippet: Human osteoblast cells (HOB) used for biocompatibility evaluation were obtained from Addexbio (Cat no: P0004010; USA).

Techniques: Comparison, Control, Cell Attachment Assay, Staining, Fluorescence, Confocal Microscopy

Journal: PLOS ONE

Article Title: Evaluation of decellularization process for developing osteogenic bovine cancellous bone scaffolds in-vitro

doi: 10.1371/journal.pone.0283922

Figure Lengend Snippet: Photomicrographs of HOB cells stained with Alizarin Red S. HOB cells showed increased mineralized nodules in osteogenic medium (OS + ) compared to cells without supplements (OS - ). More significant mineralization was seen by day 14 (a). Quantification of alizarin Red S stained mineralized nodules in human osteoblast cells seeded on DMB and DCC scaffolds grown in the presence or absence of osteogenic induction media at days 7 and 14 in comparison to cells alone is shown; p<0.001 is indicated). The absorbance of the extracted Alizarin dye was measured at 405 nm. Data presented as mean ± SEM (n = 6) (b). Quantification of Alizarin Red S staining in osteoblast cell-seeded DMB and DCC scaffolds on day 14 in the presence and absence of osteogenic supplements. Data presented as mean ± SEM (n = 4) (c); p<0.001 is indicated.

Article Snippet: Human osteoblast cells (HOB) used for biocompatibility evaluation were obtained from Addexbio (Cat no: P0004010; USA).

Techniques: Staining, Comparison

Journal: Bone

Article Title: Ex Vivo Construction of Human Primary 3D-Networked Osteocytes

doi: 10.1016/j.bone.2017.09.012

Figure Lengend Snippet: [32] (a) microfluidic perfusion device with 6 culture chambers, (b) cross-sectional view of a 3D culture chamber with the red arrows indicating the overall direction of culture medium flow through the device, (c) schematic illustration of microbeads-guided assembly, and (d) histologic image of 3D-networked osteocytes with the red arrows indicating medium flow direction with respect to the tissue sample. Scale bar: 20 µm.

Article Snippet: A human 3D bone tissue model was developed by constructing ex vivo the 3D network of osteocytes via: (1) the biomimetic assembly of primary human osteoblastic cells with 20–25 μm and BCP microbeads and (2) subsequent microfluidic perfusion culture.

Techniques:

Journal: Bone

Article Title: Ex Vivo Construction of Human Primary 3D-Networked Osteocytes

doi: 10.1016/j.bone.2017.09.012

Figure Lengend Snippet: (a) hip fragment shown as an example; (b) as-isolated cells after 4 collagenase digestion cycles; (c) proliferated osteoblastic cells after 10 days of 2D culture; (d) 3D tissue sample constructed using 20–25 µm microbeads and proliferated cells and 14 days of perfusion culture; (e) H&E histologic images showing the formation of 3D cellular network as indicated by black arrows in (f) and white arrows in (g); and (h) immunostaining for sclerostin (red). (d) –(f) from patient sample #6 and (g)–(h) from patient sample #4. Scale bar: 25 µm.

Article Snippet: A human 3D bone tissue model was developed by constructing ex vivo the 3D network of osteocytes via: (1) the biomimetic assembly of primary human osteoblastic cells with 20–25 μm and BCP microbeads and (2) subsequent microfluidic perfusion culture.

Techniques: Isolation, Construct, Immunostaining

Journal: The EMBO Journal

Article Title: TWEAK, via its receptor Fn14, is a novel regulator of mesenchymal progenitor cells and skeletal muscle regeneration

doi: 10.1038/sj.emboj.7601441

Figure Lengend Snippet: Human mesenchymal progenitor cells are a novel target cell type for TWEAK. (A) Human primary mesenchymal stem cells, skeletal muscle myoblasts, preadipocytes, chondrocytes and osteoblast precursors (Cambrex) were cultured according to the manufacturer's protocols. First-passage cells showed staining for TWEAK binding using Fc-TWEAK and for expression of Fn14 using the anti-hFn14 mAb ITEM-4. Anti-mouse and anti-human Fcs were used as negative controls, (B) NF-κB was activated in human mesenchymal stem cells (hMSCs) and osteoblast precursors (hOsteos) following 2 or 6 h of treatment with 100 ng/m TWEAK (Tw). Activation was measured using the TransAM NF-κB p65 activation assay system with cell lysates from normal and TNF-treated HeLa cells serving as negative and positive controls. The assays were carried out in triplicate and the data shown are representative of three independent experiments. (C) List of representative genes induced by TWEAK (100 ng/ml versus heat-inactivated TWEAK 100 ng/ml) in mesenchymal stem cells in low serum (LS: 0.2% FBS), moderate serum (MS: 2% FBS) and high serum (HS: 10% FBS). (D) List of some cell cycle-related genes induced by TWEAK (versus inactivated TWEAK) in mesenchymal stem cells cultured under low-serum conditions (0.2% FBS). Triplicate samples were analyzed for each condition and the fold changes were calculated using averages from triplicates. All fold changes reached statistical significance (P<0.01).

Article Snippet: These results therefore indicate that TWEAK may regulate cell fate decisions of progenitor cells. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 1 caption a7 Human mesenchymal progenitor cells are a novel target cell type for TWEAK. ( A ) Human primary mesenchymal stem cells, skeletal muscle myoblasts, preadipocytes, chondrocytes and osteoblast precursors (Cambrex) were cultured according to the manufacturer's protocols.

Techniques: Cell Culture, Staining, Binding Assay, Expressing, Activation Assay

Journal: Nanomaterials

Article Title: Synthesis of Gold Nanoparticles by Using Green Machinery: Characterization and In Vitro Toxicity

doi: 10.3390/nano11030808

Figure Lengend Snippet: Assessment of GNPs cytotoxicity and determination GNPs IC 50 on cancer cells. Normal human primary osteoblasts and human cervical cancer cell (HeLa cells) were plated overnight at a density of 1 × 10 4 cell per well in a 96-well plate at 37 °C. The normal or cancer cells were treated with different concentrations of GNPs and the in vitro cytotoxicity was evaluated using MTT assay. The inhibition percentages were calculated relative to negative control and IC 50 was the GNPs concentration, which inhibits 50% of HeLa cells. The experiment was conducted in triplicate and the data shown are the means ± standard errors.

Article Snippet: The normal human primary osteoblasts and human cervical cancer cells (HeLa) were supplied from National Centre for Cell Science (NCCS), Pune, India.

Techniques: In Vitro, MTT Assay, Inhibition, Negative Control, Concentration Assay