A Novel Formulation Based on Resveratrol and Water Extracts from Equisetum arvense, Crataegus curvisepala, Vitex agnus-castus, and Glycine max Inhibits the Gene Expression of Inflammatory and Osteoclastogenic Biomarkers on C2C12 Cells Exposed to Oxidative Stress
Abstract
:1. Introduction
2. Materials and Methods
2.1. HPLC
2.2. Colorimetric Assays
2.3. Ecotoxicological Investigation
2.3.1. Preparation of Test Sample Concentrations
2.3.2. Allelopathy Assay
2.3.3. Brine Shrimp Toxicity Bioassay
2.3.4. Daphnia magna Cardiotoxicity Assay
2.4. In Vitro Study
2.4.1. Cell Culture
2.4.2. Determination of Cell Viability
2.5. Gene Expression Analysis
2.6. In Silico Study
2.7. Statistical Analysis
3. Results
3.1. Phytochemical Analysis
3.2. Allelopathy Assay
3.3. Brine Shrimp Lethality Assay
3.4. Daphnia Magna Cardiotxocity Assay
3.5. Protective Effects on C2C12 Cells
3.6. Bioinformatics
4. Discussion
- −
- Non-toxic for LC50 > 1000 µg/mL;
- −
- Low toxicity for LC50 between 500 µg/mL and 1000 µg/mL;
- −
- Toxic for LC50 between 100 µg/mL and 500 µg/mL;
- −
- Highly toxic for LC50 < 100 µg/mL.
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Parameters | Results |
---|---|
Total phenolic content (mg GAE/gde) | 29.30 ± 1.30 |
Total flavonoid content (mg RE/gde) | 6.43 ± 0.56 |
Sample | ABTS |
---|---|
Formulation | 0.42 ± 0.06 |
Trolox | 0.03 ± 0.01 |
Seedling Length (cm) | |||
---|---|---|---|
Treatments (mg/mL) | C. inthybus | D. repens | R. sativus |
0 (Ctrl) | 28.83 ± 3.77 | 5.79 ± 0.11 | 35.89 ± 1.40 |
0.62 | 29.96 ± 4.54 | 8.69 ± 3.10 | 33.56 ± 3.62 |
1.25 | 19.38 ± 8.55 | 5.79 ± 0.77 | 35.27 ± 5.90 |
2.5 | 27.63 ± 6.78 | 8.00 ± 0.00 | 28.65 ± 1.44 |
5 | 19.90 ± 1.27 | 7.64 ± 2.73 | 29.94 ± 7.16 |
10 | 17.60 ± 3.96 | 14.81 ± 0.43 | 36.02 ± 0.45 |
Tested Extract | Concentration Range (mg/mL) | LC50 | 95% Confidence Interval | R2 | Toxicity Class | |
---|---|---|---|---|---|---|
Meyer’s classification | Clarkson’s classification | |||||
Formulation | 0.625–10 | 2.783 | 1.812–4.273 | 0.975 | Non-toxic | Non-toxic |
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Di Simone, S.C.; Acquaviva, A.; Libero, M.L.; Nilofar, N.; Tunali, F.; Gabriele, M.; Centulio, A.P.; Genovesi, G.; Ciaramellano, D.; Recinella, L.; et al. A Novel Formulation Based on Resveratrol and Water Extracts from Equisetum arvense, Crataegus curvisepala, Vitex agnus-castus, and Glycine max Inhibits the Gene Expression of Inflammatory and Osteoclastogenic Biomarkers on C2C12 Cells Exposed to Oxidative Stress. Foods 2025, 14, 896. https://doi.org/10.3390/foods14050896
Di Simone SC, Acquaviva A, Libero ML, Nilofar N, Tunali F, Gabriele M, Centulio AP, Genovesi G, Ciaramellano D, Recinella L, et al. A Novel Formulation Based on Resveratrol and Water Extracts from Equisetum arvense, Crataegus curvisepala, Vitex agnus-castus, and Glycine max Inhibits the Gene Expression of Inflammatory and Osteoclastogenic Biomarkers on C2C12 Cells Exposed to Oxidative Stress. Foods. 2025; 14(5):896. https://doi.org/10.3390/foods14050896
Chicago/Turabian StyleDi Simone, Simonetta Cristina, Alessandra Acquaviva, Maria Loreta Libero, Nilofar Nilofar, Fatma Tunali, Mariachiara Gabriele, Angelica Pia Centulio, Gianluca Genovesi, Davide Ciaramellano, Lucia Recinella, and et al. 2025. "A Novel Formulation Based on Resveratrol and Water Extracts from Equisetum arvense, Crataegus curvisepala, Vitex agnus-castus, and Glycine max Inhibits the Gene Expression of Inflammatory and Osteoclastogenic Biomarkers on C2C12 Cells Exposed to Oxidative Stress" Foods 14, no. 5: 896. https://doi.org/10.3390/foods14050896
APA StyleDi Simone, S. C., Acquaviva, A., Libero, M. L., Nilofar, N., Tunali, F., Gabriele, M., Centulio, A. P., Genovesi, G., Ciaramellano, D., Recinella, L., Leone, S., Brunetti, L., Zengin, G., Orlando, G., Menghini, L., Chiavaroli, A., & Ferrante, C. (2025). A Novel Formulation Based on Resveratrol and Water Extracts from Equisetum arvense, Crataegus curvisepala, Vitex agnus-castus, and Glycine max Inhibits the Gene Expression of Inflammatory and Osteoclastogenic Biomarkers on C2C12 Cells Exposed to Oxidative Stress. Foods, 14(5), 896. https://doi.org/10.3390/foods14050896