Journal: Oxidative Medicine and Cellular Longevity

Article Title: Extracellular Vesicles from Adipose-Derived Mesenchymal Stem Cells Downregulate Senescence Features in Osteoarthritic Osteoblasts

doi: 10.1155/2017/7197598

Figure Lengend Snippet: SA- β -Gal activity in OA osteoblasts. Cultures were treated with IL-1 β alone or in combination with MV, EX, or CM for 7 days. SA- β -Gal activity was measured by using the cellular senescence assay kit (Cell Biolabs) and expressed as relative fluorescence units (RFU). Results show mean ± SD from 4 separate experiments with cells from separate donors. ++ P < 0.01 compared to control (nonstimulated cells); ∗ P < 0.05 compared to IL-1 β .

Article Snippet: Trabecular bone samples were obtained from the femoral condyles and tibial plateaus, cut into small pieces, and subjected to enzymatic digestion with 1 mg/mL of collagenase type IA (Sigma-Aldrich) at 37°C in DMEM/HAM F-12 (Sigma-Aldrich), containing penicillin and streptomycin (1%) for 2 h. The digested tissue was cultured in osteoblast medium (Promocell, Labclinics S.A., Barcelona, Spain) in a humidified incubator with 5% CO 2 at 37°C.

Techniques: Activity Assay, Fluorescence, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Extracellular Vesicles from Adipose-Derived Mesenchymal Stem Cells Downregulate Senescence Features in Osteoarthritic Osteoblasts

doi: 10.1155/2017/7197598

Figure Lengend Snippet: Release of inflammatory mediators. IL-6 (a) and TNF α (b) were measured by ELISA; PGE 2 (c) was measured by radioimmunoassay in cell culture supernatants of OA osteoblasts. Cultures were treated with IL-1 β and/or MV, EX, or CM for 24 h. Results are expressed as mean ± SD from 4 separate experiments with cells from separate donors. ++ P < 0.01 compared to control (nonstimulated cells); ∗∗ P < 0.01 compared to IL-1 β .

Article Snippet: Trabecular bone samples were obtained from the femoral condyles and tibial plateaus, cut into small pieces, and subjected to enzymatic digestion with 1 mg/mL of collagenase type IA (Sigma-Aldrich) at 37°C in DMEM/HAM F-12 (Sigma-Aldrich), containing penicillin and streptomycin (1%) for 2 h. The digested tissue was cultured in osteoblast medium (Promocell, Labclinics S.A., Barcelona, Spain) in a humidified incubator with 5% CO 2 at 37°C.

Techniques: Enzyme-linked Immunosorbent Assay, RIA Assay, Cell Culture, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Extracellular Vesicles from Adipose-Derived Mesenchymal Stem Cells Downregulate Senescence Features in Osteoarthritic Osteoblasts

doi: 10.1155/2017/7197598

Figure Lengend Snippet: Release of IL-10 by OA osteoblasts. IL-10 was measured by ELISA in cell culture supernatants. Cultures were treated with IL-1 β and/or MV, EX, or CM for 24 h. Results are expressed as mean ± SD from 5 separate experiments with cells from separate donors. ∗∗ P < 0.01 compared to IL-1 β .

Article Snippet: Trabecular bone samples were obtained from the femoral condyles and tibial plateaus, cut into small pieces, and subjected to enzymatic digestion with 1 mg/mL of collagenase type IA (Sigma-Aldrich) at 37°C in DMEM/HAM F-12 (Sigma-Aldrich), containing penicillin and streptomycin (1%) for 2 h. The digested tissue was cultured in osteoblast medium (Promocell, Labclinics S.A., Barcelona, Spain) in a humidified incubator with 5% CO 2 at 37°C.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Extracellular Vesicles from Adipose-Derived Mesenchymal Stem Cells Downregulate Senescence Features in Osteoarthritic Osteoblasts

doi: 10.1155/2017/7197598

Figure Lengend Snippet: Quantification of HNE-protein adducts in OA osteoblasts. HNE-protein adducts were measured by ELISA in cellular extracts. Cultures were treated with IL-1 β alone or in combination with MV, EX, or CM for 24 h. Results are expressed as mean ± SD from 4 separate experiments with cells from separate donors. ++ P < 0.01 compared to control (nonstimulated cells); ∗ P < 0.05 and ∗∗ P < 0.01 compared to IL-1 β .

Article Snippet: Trabecular bone samples were obtained from the femoral condyles and tibial plateaus, cut into small pieces, and subjected to enzymatic digestion with 1 mg/mL of collagenase type IA (Sigma-Aldrich) at 37°C in DMEM/HAM F-12 (Sigma-Aldrich), containing penicillin and streptomycin (1%) for 2 h. The digested tissue was cultured in osteoblast medium (Promocell, Labclinics S.A., Barcelona, Spain) in a humidified incubator with 5% CO 2 at 37°C.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Extracellular Vesicles from Adipose-Derived Mesenchymal Stem Cells Downregulate Senescence Features in Osteoarthritic Osteoblasts

doi: 10.1155/2017/7197598

Figure Lengend Snippet: Analysis of mitochondrial membrane potential in OA osteoblasts. Analysis was performed by flow cytometry using the probe JC-1. Representative images (a); green/red fluorescence ratio (b). Cultures were treated with IL-1 β alone or in combination with MV, EX, or CM for 24 h. Results are expressed as mean ± SD from 3 separate experiments with cells from separate donors. ++ P < 0.01 compared to control (nonstimulated cells); ∗ P < 0.05 and ∗∗ P < 0.01 compared to IL-1 β .

Article Snippet: Trabecular bone samples were obtained from the femoral condyles and tibial plateaus, cut into small pieces, and subjected to enzymatic digestion with 1 mg/mL of collagenase type IA (Sigma-Aldrich) at 37°C in DMEM/HAM F-12 (Sigma-Aldrich), containing penicillin and streptomycin (1%) for 2 h. The digested tissue was cultured in osteoblast medium (Promocell, Labclinics S.A., Barcelona, Spain) in a humidified incubator with 5% CO 2 at 37°C.

Techniques: Membrane, Flow Cytometry, Fluorescence, Control

Journal: Scientific Reports

Article Title: Positive impact of Platelet-rich plasma and Platelet-rich fibrin on viability, migration and proliferation of osteoblasts and fibroblasts treated with zoledronic acid

doi: 10.1038/s41598-019-43798-z

Figure Lengend Snippet: Preinvestigation of PRP and PRF for in-vitro addition. The effect of PRP and PRF were examined in different concentrations to fibroblasts ( A ) and osteoblasts ( B ) by real- time cell analysis.

Article Snippet: The fibroblast and osteoblast growth medium (FGM and OGM; both from Provitro®; Berlin Germany) were each supplemented with 10% fetal bovine serum (FBS, Provitro® GmbH) and 1% antibiotics (100 U/ml penicillin G, 100 μg/ml streptomycin).

Techniques: In Vitro, Cell Analysis

Journal: Scientific Reports

Article Title: Positive impact of Platelet-rich plasma and Platelet-rich fibrin on viability, migration and proliferation of osteoblasts and fibroblasts treated with zoledronic acid

doi: 10.1038/s41598-019-43798-z

Figure Lengend Snippet: Representative photographs of cell migration using the scratch assay. The effect of ZA, PRP, PRF and their combination on the migration of fibroblasts ( A ) and osteoblasts ( B ) were examined (PC = positive control; NC = negative control; ZA = zoledronic acid).

Article Snippet: The fibroblast and osteoblast growth medium (FGM and OGM; both from Provitro®; Berlin Germany) were each supplemented with 10% fetal bovine serum (FBS, Provitro® GmbH) and 1% antibiotics (100 U/ml penicillin G, 100 μg/ml streptomycin).

Techniques: Migration, Wound Healing Assay, Positive Control, Negative Control

Journal: Scientific Reports

Article Title: Positive impact of Platelet-rich plasma and Platelet-rich fibrin on viability, migration and proliferation of osteoblasts and fibroblasts treated with zoledronic acid

doi: 10.1038/s41598-019-43798-z

Figure Lengend Snippet: Percentage of closure of the scratch area in fibroblasts ( A ) and osteoblasts ( B ) in the different groups (1.0 = 100% coverage). Significant difference ( p < 0.05) is marked with *.

Article Snippet: The fibroblast and osteoblast growth medium (FGM and OGM; both from Provitro®; Berlin Germany) were each supplemented with 10% fetal bovine serum (FBS, Provitro® GmbH) and 1% antibiotics (100 U/ml penicillin G, 100 μg/ml streptomycin).

Techniques:

Journal: Scientific Reports

Article Title: Positive impact of Platelet-rich plasma and Platelet-rich fibrin on viability, migration and proliferation of osteoblasts and fibroblasts treated with zoledronic acid

doi: 10.1038/s41598-019-43798-z

Figure Lengend Snippet: Effects of PRP, PRF, ZA and their combinations on the viability of human gingival fibroblasts ( A ) and osteoblasts ( B ) assessed by MTT assay. Significant difference ( p < 0.05) is marked with *.

Article Snippet: The fibroblast and osteoblast growth medium (FGM and OGM; both from Provitro®; Berlin Germany) were each supplemented with 10% fetal bovine serum (FBS, Provitro® GmbH) and 1% antibiotics (100 U/ml penicillin G, 100 μg/ml streptomycin).

Techniques: MTT Assay

Journal: Scientific Reports

Article Title: Positive impact of Platelet-rich plasma and Platelet-rich fibrin on viability, migration and proliferation of osteoblasts and fibroblasts treated with zoledronic acid

doi: 10.1038/s41598-019-43798-z

Figure Lengend Snippet: Cell proliferation assessed by real-time cell analysis for fibroblasts ( A ) and osteoblasts ( B ) in the different groups over a period of 72 h. The values are the median of the measured cell index. Significant difference ( p < 0.05) is marked with *.

Article Snippet: The fibroblast and osteoblast growth medium (FGM and OGM; both from Provitro®; Berlin Germany) were each supplemented with 10% fetal bovine serum (FBS, Provitro® GmbH) and 1% antibiotics (100 U/ml penicillin G, 100 μg/ml streptomycin).

Techniques: Cell Analysis

Journal: British Journal of Cancer

Article Title: A mechanism of resistance to TRAIL/Apo2L-induced apoptosis of newly established glioma cell line and sensitisation to TRAIL by genotoxic agents

doi: 10.1038/sj.bjc.6600666

Figure Lengend Snippet: Sensitivity of osteosarcoma, fibrosarcoma, glioma cell lines, human normal osteoblasts and fibroblasts to TRAIL. ( A ) After 48 h of culture with 500 ngml −1 of TRAIL, the cell viability of HOS, MG63, NY, SaOS and osteoblasts was determined by crystal violet assay as described in Materials and Methods. The means and standard deviations of three independent experiments are shown. Statistical significance: * P <0.01 compared with controls. ( B ) After 48 h of culture with 2 μ gml −1 of TRAIL, the cell viability of HT1080 and fibroblasts was determined by crystal violet assay. The means and s.d.'s of five independent experiments are shown. Statistical significance: * P <0.01 compared with controls. ( C ) After 48 h of culture with 2 μ gml −1 of TRAIL, the cell viability of T98G, A172, KG-1-C, and #63 was determined by crystal violet assay. The means and s.d.'s of five independent experiments are shown. Statistical significance: * P <0.01 compared with controls.

Article Snippet: Human normal osteoblasts were purchased from Sanko-junyaku (Tokyo, Japan) and maintained with Osteoblast Growth Medium (Osteoblast Growth Medium Bullet Kit).

Techniques: Crystal Violet Assay