Impact of Ilex paraguariensis extract on osteoblastic phenotype expression

The use of dietary supplements and the functionalization of biomaterials with natural compounds have been explored to enhance bone regeneration. While Ilex paraguariensis St. Hil (yerba mate) has shown benefits in various tissues, its effects on bone tissue remain underexplored. This study evaluated the impact of Ilox^® Micro extract on in vitro osteoblastic differentiation.

To this end, osteoblasts from the UMR-106 cell line were exposed to 0 (control), 10, 30, and 60 µg/mL of lyophilized Ilox^® Micro extract. The antioxidant capacity and total phenolic content were determined using the colorimetric Folin-Ciocalteu method. Cell viability was assessed using the Live/Dead kit (3 days) and the resazurin reduction method (3 and 5 days). The expression of Runx-2, Sp7, Col1a1, Alpl, Ibsp and Bglap genes was quantified by real-time PCR (5 days), and in situ alkaline phosphatase (ALP) activity and mineralized matrix were evaluated using Fast Red (5 days) and Alizarin Red (7 days) staining, respectively. Quantitative data were analyzed using ANOVA (α = 5%). Live/Dead staining demonstrated that adhered and spread cells were predominantly viable, with cell density increasing proportionally to the Ilox^® Micro extract concentration. The resazurin reduction assay indicated higher values at 30 and 60 µg/mL. While Sp7, Col1a1, Alpl, and Ibsp showed no significant differences, Runx2 and Bglap were upregulated at 60 µg/mL. Additionally, 30 µg/mL exhibited greater ALP staining, and a trend toward increased mineralization was observed at 10 and 30 µg/mL.

Based on the obtained results, it can be concluded that Ilox^® Micro extract can positively modulate some key phenomena associated with the acquisition of the osteoblastic phenotype in vitro.