Journal: Food Science and Biotechnology

Article Title: Anti-inflammatory and in vitro bone formation effects of Garcinia mangostana L. and propolis extracts

doi: 10.1007/s10068-019-00697-3

Figure Lengend Snippet: Cell viability of immortalized human gingival fibroblasts (hTERT-hNOF) after treatment with mangosteen and/or propolis extracts. hTERT-hNOF cells were seeded onto a 24-well plate at a density of 5 × 104 cells/well and cultured overnight. After reaching 80% confluence, cells were treated with mangosteen and/or propolis extracts alone or in combination. After cells were cultured for 24 h, culture medium was removed and a medium containing 10% of 5 mg/ml methylthiazolyldiphenyl-tetrazolium bromide solution was added to each well followed by incubation for 4 h. After dissolving in isopropanol, the absorbance was measured at 570 nm using an ELISA reader and results were analyzed. Each reaction was repeated three times. *p < 0.05

Article Snippet: ELISA was performed using human runx2 ELISA kit (Biomatik, Wilmington, DE, USA), human osterix ELISA kit (Biomatik), human type I collagen ELISA kit (Biomatik), and human osteocalcin ELISA kit (Biomatik).

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Food Science and Biotechnology

Article Title: Anti-inflammatory and in vitro bone formation effects of Garcinia mangostana L. and propolis extracts

doi: 10.1007/s10068-019-00697-3

Figure Lengend Snippet: Effect of mangosteen and/or propolis extracts on in vitro bone formation in human osteoblast-like cell line MG-63 cells. MG63 cells were seeded onto 12-well plates at a density of 1 × 105 cells/well and cultured overnight. For differentiation, MG-63 cells were cultured in a medium supplemented with 50 μg/ml ascorbic acid, 10 mM β-glycerophosphate, and mangosteen and/or propolis extracts for 28 days. (A) Alizarin red staining was performed according to the method of Reinholz et al. (2000) to observe minerals produced in the extracellular matrix. First, the medium was removed. Cells were then washed with PBS and fixed with 2.5% glutaraldehyde at room temperature for 15 min. These cells were washed with PBS again and stained with 2% Alizarin red solution for 10 min. (A) After washing with distilled water, calcification was observed based on the degree of red staining after taking a photo. (B) After drying, the red staining was dissolved in a resolving solution (20% methanol and 10% acetic acid) and absorbance was measured at 450 nm with an ELISA reader. Results were then analyzed. Each reaction was repeated three times. *p < 0.05

Article Snippet: ELISA was performed using human runx2 ELISA kit (Biomatik, Wilmington, DE, USA), human osterix ELISA kit (Biomatik), human type I collagen ELISA kit (Biomatik), and human osteocalcin ELISA kit (Biomatik).

Techniques: In Vitro, Cell Culture, Staining, Produced, Enzyme-linked Immunosorbent Assay

Journal: Food Science and Biotechnology

Article Title: Anti-inflammatory and in vitro bone formation effects of Garcinia mangostana L. and propolis extracts

doi: 10.1007/s10068-019-00697-3

Figure Lengend Snippet: Effect of mangosteen and/or propolis extracts on expression of alkaline phosphatase (ALP) in human osteoblast-like cell line MG-63. MG63 cells were seeded onto 6-well plates at a density of 3 × 105 cells/well and incubated at 37 °C with 5% CO2 and 100% humidity in a medium supplemented with 50 μg/ml ascorbic acid, 10 mM β-glycerophosphate, and mangosteen and/or propolis extracts for 1, 4, 7, and 14 days. Cells were lysed with cold Triton X-100 and centrifuged. Supernatants were incubated with 0.1 M glycine–NaOH buffer (pH 10.4) and 15 mM p-nitrophenyl phosphate at 37 °C for 30 min. After the same amount of 1.2 N NaOH was added to stop the reaction, the mixture was transferred to an ELISA reader and absorbance was measured at 405 nm. Protein concentration was measured using a BCA Protein Assay kit. Enzyme activity was calculated as μmole p-nitrophenol/min/μg protein. Each reaction was repeated three times. *p < 0.05

Article Snippet: ELISA was performed using human runx2 ELISA kit (Biomatik, Wilmington, DE, USA), human osterix ELISA kit (Biomatik), human type I collagen ELISA kit (Biomatik), and human osteocalcin ELISA kit (Biomatik).

Techniques: Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Protein Concentration, Bicinchoninic Acid Protein Assay, Activity Assay