Journal: Open Veterinary Journal

Article Title: Generation and characterization of mesenchymal stem cells from the affected femoral heads of dogs with Legg Calvé Perthes disease

doi: 10.5455/OVJ.2024.v14.i5.12

Figure Lengend Snippet: Trilineage differentiation potency. Canine LCPD-affected femoral head-derived adherent cells (A-C) and BMMSCs (D-F) differentiated into adipocytes (A and D), osteoblasts (B and E), and chondrocytes (C and F). (A and D): Sudan III staining, (B and E): Alizarin Red staining, and (C and F): Alcian blue staining. Scale bars = 100 μm or 1 mm.

Article Snippet: Canine osteoblast differentiation medium (Cell Applications, Inc.) was used for osteoblast differentiation, and the same static culture was performed for 2 weeks.

Techniques: Derivative Assay, Staining

Journal: Communications Biology

Article Title: mTORC1 impedes osteoclast differentiation via calcineurin and NFATc1

doi: 10.1038/s42003-018-0028-4

Figure Lengend Snippet: Late mTORC1 deletion in myeloid lineage enhances osteoclastogenesis. Analyses of Raptor fl/fl -Lyz-Cre cKO mice and littermate controls. a , b Expression of osteoclast markers ( a ) and osteoclastogenic transcription factors ( b ) in bone marrow osteoclast differentiation cultures ( n = 6). c Representative images of TRAP-stained osteoclast differentiation cultures. d Osteoclast resorptive activity measured by calcium release from bone plates ( n = 16). e Osteoclast precursor proliferation by BrdU incorporation ( n = 10). f , g mTORC1 signaling in bone marrow osteoclast differentiation cultures from Raptor fl/fl -Lyz mice or littermate controls 50 h after RANKL treatment. f Phosphorylation of mTOR; g Phosphorylation of S6K1. p/T, ratio of phosphorylated/total protein. h – t Bone phenotype in 3-month-old male mice. h Serum CTX-1 bone resorption marker ( n = 8). i Serum P1NP bone formation marker ( n = 8). j Representative μCT images of the trabecular bone of the tibial metaphysis (top) and the entire proximal tibia (bottom). k – p Quantification of trabecular bone volume and architecture in proximal tibiae by μCT ( n = 8–10). k BV/TV bone volume/tissue volume ratio; l BS bone surface; m Tb.N trabecular number; n Conn. D. connectivity density; o Tb.Sp trabecular separation; p SMI structure model index. q – t Bone histomorphometry of distal femurs ( n = 6). Oc.S osteoclast surface, B.S bone surface, Oc.N osteoclast number, Ob.S osteoblast surface, Ob.N osteoblast number. Error bars, SEM; *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.001; n.s. non-significant. R raptor. Full-size scans of immunoblots are shown in Supplementary Fig.

Article Snippet: Osteoblast precursors were expanded for 4 days in Mesenchymal Stem Cell (MSC) media using a Mouse MesenCult Proliferation Kit (StemCell Technologies) before the addition of osteoblast differentiation cocktail (StemCell Technologies).

Techniques: Expressing, Staining, Activity Assay, BrdU Incorporation Assay, Phospho-proteomics, Marker, Western Blot

Journal: Communications Biology

Article Title: mTORC1 impedes osteoclast differentiation via calcineurin and NFATc1

doi: 10.1038/s42003-018-0028-4

Figure Lengend Snippet: Constitutive mTORC1 activation in myeloid lineage impedes osteoclast differentiation. Analyses of Tsc1 fl/fl -Lyz-Cre cKO mice and littermate controls. a , b Expression of osteoclast markers ( a ) and osteoclastogenic transcription factors ( b ) in bone marrow osteoclast differentiation cultures ( n = 6). c Representative images of TRAP-stained osteoclast differentiation cultures. d Osteoclast resorptive activity measured by calcium release from bone plates ( n = 16). e Osteoclast precursor proliferation by BrdU incorporation ( n = 10). f , g mTORC1 signaling in bone marrow osteoclast differentiation cultures 60 h after RANKL treatment. f Phosphorylation of mTOR; g Phosphorylation of S6K1. p/T ratio of phosphorylated/total protein. h – r Bone phenotype in 2-month-old male mice. h Serum CTX-1 bone resorption marker ( n = 8). i Serum P1NP bone formation marker ( n = 8). j Bone histomorphometry of distal femurs ( n = 6). Oc.S osteoclast surface, B.S bone surface, Oc.N osteoclast number. k Bone histomorphometry of distal femurs ( n = 6). Ob.S osteoblast surface, B.S bone surface, Ob.N osteoblast number. l Representative μCT images of the trabecular bone of the tibial metaphysis (top) and the entire proximal tibia (bottom). m – r Quantification of trabecular bone volume and architecture in proximal tibiae by μCT ( n = 8). m BV/TV bone volume/tissue volume ratio, n BS bone surface, o Tb.N trabecular number, p Tb.Sp trabecular separation, q Conn. D. connectivity density, r SMI structure model index. Error bars, SEM; * p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.001; n.s. non-significant. T Tsc1. Full-size scans of immunoblots are shown in Supplementary Fig.

Article Snippet: Osteoblast precursors were expanded for 4 days in Mesenchymal Stem Cell (MSC) media using a Mouse MesenCult Proliferation Kit (StemCell Technologies) before the addition of osteoblast differentiation cocktail (StemCell Technologies).

Techniques: Activation Assay, Expressing, Staining, Activity Assay, BrdU Incorporation Assay, Phospho-proteomics, Marker, Western Blot

Journal: Advanced Healthcare Materials

Article Title: Biological Characterization of Ti6Al4V Additively Manufactured Surfaces: Comparison Between Ultrashort Laser Texturing and Conventional Post‐Processing

doi: 10.1002/adhm.202402873

Figure Lengend Snippet: Representative fluorescent images of osteoblast stained with phalloidin following adhesion onto the different surfaces after 24 h. Images were acquired at 20X magnification with an inverted microscope equipped with an epifluorescence setup (Eclipse TiU, NIKON Europe BV, NITAL SpA, Milano, Italy) (Scale bar: 100 µm).

Article Snippet: The cells were cultivated and expanded in osteoblast basal medium (OBM Osteoblast Growth and Differentiation Basal Medium; LONZA), supplemented with the appropriate additives (OGM Osteoblasts Growth SingleQuots kit, LONZA), 10% fetal bovine serum (FBS, EUROCLONE, Pero, Milano, Italy), 100 U mL −1 penicillin, 100 μg mL −1 streptomycin, (SIGMA, St. Louis, MO) under standard conditions (37 °C, 5%CO 2 /95%air, humidified atmosphere).

Techniques: Staining, Inverted Microscopy

Journal: Advanced Healthcare Materials

Article Title: Biological Characterization of Ti6Al4V Additively Manufactured Surfaces: Comparison Between Ultrashort Laser Texturing and Conventional Post‐Processing

doi: 10.1002/adhm.202402873

Figure Lengend Snippet: Morphometric descriptors of osteoblast shape in response to XZ surfaces sandblasted with corundum (SB‐C), zirconia (SB‐Z), and laser textured (LT).

Article Snippet: The cells were cultivated and expanded in osteoblast basal medium (OBM Osteoblast Growth and Differentiation Basal Medium; LONZA), supplemented with the appropriate additives (OGM Osteoblasts Growth SingleQuots kit, LONZA), 10% fetal bovine serum (FBS, EUROCLONE, Pero, Milano, Italy), 100 U mL −1 penicillin, 100 μg mL −1 streptomycin, (SIGMA, St. Louis, MO) under standard conditions (37 °C, 5%CO 2 /95%air, humidified atmosphere).

Techniques:

Journal: Advanced Healthcare Materials

Article Title: Biological Characterization of Ti6Al4V Additively Manufactured Surfaces: Comparison Between Ultrashort Laser Texturing and Conventional Post‐Processing

doi: 10.1002/adhm.202402873

Figure Lengend Snippet: Metabolic activity was measured with Alamar Blue assay at 3, 7, and 14 days of culture of osteoblasts seeded onto different surfaces finishing conditions realized onto titanium samples with a) XY orientation or b) XZ orientation. The results are given as relative fluorescent units (RFU) and values are reported as mean and [95% CI] obtained from four replicate materials. Statistical comparison is reported in the graphs: among experimental times within the same material three different symbols were used: # for SB‐C; § for SB‐Z and ° for LT (for example # , p < 0.05; ## , p < 0.005; ### , p < 0.001) and among experimental materials within the same experimental time ( * , p < 0.05; ** , p < 0.005; *** , p < 0.001).

Article Snippet: The cells were cultivated and expanded in osteoblast basal medium (OBM Osteoblast Growth and Differentiation Basal Medium; LONZA), supplemented with the appropriate additives (OGM Osteoblasts Growth SingleQuots kit, LONZA), 10% fetal bovine serum (FBS, EUROCLONE, Pero, Milano, Italy), 100 U mL −1 penicillin, 100 μg mL −1 streptomycin, (SIGMA, St. Louis, MO) under standard conditions (37 °C, 5%CO 2 /95%air, humidified atmosphere).

Techniques: Activity Assay, Alamar Blue Assay, Comparison

Journal: Advanced Healthcare Materials

Article Title: Biological Characterization of Ti6Al4V Additively Manufactured Surfaces: Comparison Between Ultrashort Laser Texturing and Conventional Post‐Processing

doi: 10.1002/adhm.202402873

Figure Lengend Snippet: Representative photomicrographs acquired using scanning electron microscopy showing the osteoblast spreading at 7 days onto the two orientations surface finished with two techniques represented by sandblasting with corundum (SB‐C)(a)(d) or zirconia (SB‐Z) (b)(e) and laser texturing (LT)(c)(f).

Article Snippet: The cells were cultivated and expanded in osteoblast basal medium (OBM Osteoblast Growth and Differentiation Basal Medium; LONZA), supplemented with the appropriate additives (OGM Osteoblasts Growth SingleQuots kit, LONZA), 10% fetal bovine serum (FBS, EUROCLONE, Pero, Milano, Italy), 100 U mL −1 penicillin, 100 μg mL −1 streptomycin, (SIGMA, St. Louis, MO) under standard conditions (37 °C, 5%CO 2 /95%air, humidified atmosphere).

Techniques: Electron Microscopy

Journal: Advanced Healthcare Materials

Article Title: Biological Characterization of Ti6Al4V Additively Manufactured Surfaces: Comparison Between Ultrashort Laser Texturing and Conventional Post‐Processing

doi: 10.1002/adhm.202402873

Figure Lengend Snippet: TGF‐β1 and BMP‐2 gene expression at 7 and 14 days of culture of osteoblasts seeded onto different surfaces finishing conditions realized onto titanium samples with XY orientation (a and c) or XZ orientation (b and d). The results show the mean and 95% CI obtained from three replicate materials. Statistical analysis is reported in the graphs: between experimental times in the same material, three different symbols were used: # for SB‐C; § for SB‐Z, and ° for LT (for example ### , p < 0.001; ## , p < 0.005; # , p < 0.05) and among experimental materials in the same experimental time ( *** , p < 0.001; ** , p < 0.005; * , p < 0.05).

Article Snippet: The cells were cultivated and expanded in osteoblast basal medium (OBM Osteoblast Growth and Differentiation Basal Medium; LONZA), supplemented with the appropriate additives (OGM Osteoblasts Growth SingleQuots kit, LONZA), 10% fetal bovine serum (FBS, EUROCLONE, Pero, Milano, Italy), 100 U mL −1 penicillin, 100 μg mL −1 streptomycin, (SIGMA, St. Louis, MO) under standard conditions (37 °C, 5%CO 2 /95%air, humidified atmosphere).

Techniques: Gene Expression

Journal: Advanced Healthcare Materials

Article Title: Biological Characterization of Ti6Al4V Additively Manufactured Surfaces: Comparison Between Ultrashort Laser Texturing and Conventional Post‐Processing

doi: 10.1002/adhm.202402873

Figure Lengend Snippet: ALPL, COL1A1 , and SSP1 gene expression at 7 and 14 days of culture of osteoblasts seeded onto different surfaces finishing conditions realized onto titanium samples with XY orientation (a‐c) or XZ orientation (d‐f). The results show the mean and 95% CI obtained from three replicate materials. Statistical analysis is reported in the graphs: between experimental times in the same material three different symbols were used: # for SB‐C; § for SB‐Z and ° for LT (for example: ### , p < 0.001; ## , p < 0.005; # , p < 0.05) and among experimental materials in the same experimental time ( *** : p < 0.001; ** : p < 0.005; * : p < 0.05).

Article Snippet: The cells were cultivated and expanded in osteoblast basal medium (OBM Osteoblast Growth and Differentiation Basal Medium; LONZA), supplemented with the appropriate additives (OGM Osteoblasts Growth SingleQuots kit, LONZA), 10% fetal bovine serum (FBS, EUROCLONE, Pero, Milano, Italy), 100 U mL −1 penicillin, 100 μg mL −1 streptomycin, (SIGMA, St. Louis, MO) under standard conditions (37 °C, 5%CO 2 /95%air, humidified atmosphere).

Techniques: Gene Expression

Journal: Advanced Healthcare Materials

Article Title: Biological Characterization of Ti6Al4V Additively Manufactured Surfaces: Comparison Between Ultrashort Laser Texturing and Conventional Post‐Processing

doi: 10.1002/adhm.202402873

Figure Lengend Snippet: Quantitative results of ELISA assays performed onto osteoblast's supernatant in response to XZ surfaces sandblasted with corundum (SB‐C), zirconia (SB‐Z), and laser textured (LT).

Article Snippet: The cells were cultivated and expanded in osteoblast basal medium (OBM Osteoblast Growth and Differentiation Basal Medium; LONZA), supplemented with the appropriate additives (OGM Osteoblasts Growth SingleQuots kit, LONZA), 10% fetal bovine serum (FBS, EUROCLONE, Pero, Milano, Italy), 100 U mL −1 penicillin, 100 μg mL −1 streptomycin, (SIGMA, St. Louis, MO) under standard conditions (37 °C, 5%CO 2 /95%air, humidified atmosphere).

Techniques: Enzyme-linked Immunosorbent Assay