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Potential of delphinidin-3-rutinoside extracted from Solanum melongena L. as promoter of osteoblastic MC3T3-E1 function and antagonist of oxidative damage

European Journal of Nutrition volume 58, pages 1019–1032 (2019)Cite this article

Abstract

Purpose

Increasing evidence suggests the potential use of natural antioxidant compounds in the prevention/treatment of osteoporosis. This study was undertaken to investigate the effects of purified delphinidin-3-rutinoside (D3R), isolated from Solanum melongena L., on osteoblast viability and differentiation in basal conditions and its ability to protect MC3T3-E1 cells against oxidative damage induced by tert-butyl hydroperoxide (t-BHP).

Methods

MC3T3-E1 osteoblastic cells were treated with D3R (10−11–10−5 M for 24 h), followed by treatment with t-BHP (250 µM for 3 h). To test cell viability, MTT test was performed. Apoptotic cells were stained with Hoechst-33258 dye. Cytoskeleton rearrangement was stained with FICT-labelled phalloidin. Intracellular ROS production was measured using dichlorofluorescein CM-DCFA. The reduced glutathione to oxidized glutathione ratio (GSH/GSSG) contents was measured according to the OPT fluorimetric assay.

Results

D3R (10−9 M) significantly increases viability of MC3T3-E1 cells and promotes osteoblast differentiation by increasing the expression of type I collagen, alkaline phosphatase and osteocalcin. Pre-treatment with D3R (10−9 M) significantly prevented t-BHP-induced osteoblastic dysfunction and changes in the cytoskeleton organization by decreasing intracellular ROS and preventing the reduction in GSH/GSSG. D3R did not significantly modify the expression of Osteoprotegerin/RANKL system activated by t-BHP suggesting a lack of effect of D3R on osteoblast/osteoclast crosstalk. D3R protective effects against t-BHP-induced osteoblastic dysfunction were mediated by the PI3K/Akt pathway since they were completely prevented by LY294002, a PI3K/Akt specific inhibitor.

Conclusions

These findings indicate that D3R protects MC3T3-E1 cells from oxidative damage and suggest the potential utility of dietary D3R supplement to prevent osteoblast dysfunction in age-related osteoporosis.

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Acknowledgements

This work was supported by funds from PROGETTO CARIPLO GIOVANI 2015−0834 to Lavinia Casati. The authors thank the expertise and technical support of Dr. Giuseppe L. Rotino (CREA-ORL, Montanaso Lombardo) for providing the aubergine fruits and Prof. Giovanna Speranza (Dipartimento di Chimica, Università degli Studi di Milano) for the analysis by 1H-NMR of D3R crystals purity.

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  1. Department of Medical Biotechnology and Translational Medicine, Università degli Studi di Milano, Via Vanvitelli, 32, 20129, Milano, Italy

    Lavinia Casati, Francesca Pagani & Valeria Sibilia

  2. Research Centre for Engineering and Agro-Food Processing (CREA-IT), Via Venezian 26, 20133, Milano, Italy

    Marta Fibiani & Roberto Lo Scalzo

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  1. Lavinia Casati
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  2. Francesca Pagani
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Correspondence to Valeria Sibilia.

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Casati, L., Pagani, F., Fibiani, M. et al. Potential of delphinidin-3-rutinoside extracted from Solanum melongena L. as promoter of osteoblastic MC3T3-E1 function and antagonist of oxidative damage. Eur J Nutr 58, 1019–1032 (2019). https://doi.org/10.1007/s00394-018-1618-0

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Keywords

  • Delphinidin-3-rutinoside
  • Functional food component
  • Oxidative stress
  • Osteoblastic MC3T3-E1 cells