Journal: Molecules

Article Title: Silver Fir ( Abies alba L.) Polyphenolic Extract Shows Beneficial Influence on Chondrogenesis In Vitro under Normal and Inflammatory Conditions

doi: 10.3390/molecules25204616

Figure Lengend Snippet: Chondrogenesis under non-inflammatory conditions. Chondrogenic differentiation was determined using the micromass assay. ( A ) Primary human (h)MSCs from patients with hip osteoarthritis ( n = 3) and donors post mortem without musculoskeletal disorders (healthy donors, n = 2) were used to determine the effects of the polyphenolic extracts and polyphenols on chondrogenesis. Micromasses were treated with the polyphenolic extracts and polyphenols, with the addition of TGF-β1 for 7 days. Control micromasses received a medium without TGF-β1. Alcian blue staining was performed to visualize the glycosaminoglycans formed. The absorbance of the extracted Alcian blue was then measured at 630 nm, as the measure of chondrogenesis. Significant benefits were seen for Belinal over the control and resveratrol treatments, as indicated (one-way ANOVA with Bonferroni correction). ( B ) Representative scans of the Alcian blue-stained glycosaminoglycans in the micromasses. B, Belinal; P, Pycnogenol; R, resveratrol; Q, quercetin.

Article Snippet: The micromasses were fixed using ice-cold methanol (Merck Milipore, Burlington, MA, USA) for 30 min at −20 °C.

Techniques: Control, Staining

Journal: Molecules

Article Title: Silver Fir ( Abies alba L.) Polyphenolic Extract Shows Beneficial Influence on Chondrogenesis In Vitro under Normal and Inflammatory Conditions

doi: 10.3390/molecules25204616

Figure Lengend Snippet: Chondrogenesis under inflammatory conditions. ( A ) Cell proliferation screening to determine the optimal lipopolysaccharide (LPS) concentration for further experiments, using the MTS assay. Serial dilutions of LPS (1, 0.1, 0.01, 0.001 µg/mL) were added to the primary hMSCs for 18 h, to simulate inflammatory conditions. After 18 h, the medium was changed and the polyphenolic extracts and polyphenols were added at 375 mg/mL. No significant differences in viability (as measured at 490 nm in the MTS assay) were seen between the serial dilutions of LPS. LPS at 0.01 µg/mL was used for further experiments based on previous studies [ , ]. Each concentration of LPS for each treatment was tested in duplicate. ( B ) Chondrogenic differentiation was determined using the hMSCs from patients with hip osteoarthritis ( n = 3) in the micromass assay. Micromasses were pretreated with 0.01 µg/mL LPS for 18 h, and then treated with the polyphenolic extracts and polyphenols, with the addition of TGF-β1 for 7 days. After 7 days, Alican blue staining was performed as the measure of chondrogenesis. Belinal showed significantly greater absorbance of extracted Alcian blue in comparison with the controls that only received cell medium. B, Belinal; P, Pycnogenol; R, resveratrol; Q, quercetin.

Article Snippet: The micromasses were fixed using ice-cold methanol (Merck Milipore, Burlington, MA, USA) for 30 min at −20 °C.

Techniques: Concentration Assay, MTS Assay, Staining, Comparison