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Article

Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese

by
Cristina Martelli
1,†,
Luisa Ottobrini
1,2,†,
Anita Ferraretto
3,
Paola Bendinelli
3,
Stefano Cattaneo
4,
Fabio Masotti
4,
Milda Stuknytė
5,
Margherita Dall’Asta
6,
Angelo Del Sole
7,
Ivano De Noni
4,* and
Filippo Rossi
6
1
Department of Pathophysiology and Transplantation, University of Milan, 20054 Milan, Italy
2
Bioimaging and Complex Biological Systems (IBSBC), National Research Council (CNR), 20054 Milan, Italy
3
Department of Biomedical Sciences for Health, University of Milan, 20133 Milan, Italy
4
Department of Food, Environmental and Nutritional Sciences, University of Milan, 20133 Milan, Italy
5
Unitech COSPECT—COmprehensive Substances characterization via advanced sPECTtroscopy, University Technological Platform, University of Milan, 20133 Milan, Italy
6
Department of Animal Science, Food and Nutrition, Università Cattolica del Sacro Cuore, 29122 Piacenza, Italy
7
Department of Health Sciences, University of Milan, 20146 Milan, Italy
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Foods 2025, 14(2), 273; https://doi.org/10.3390/foods14020273
Submission received: 8 December 2024 / Revised: 7 January 2025 / Accepted: 14 January 2025 / Published: 16 January 2025
(This article belongs to the Special Issue Reviews and Research on New Developments and Advances in Dairy Science and Technology)
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Abstract

Grana Padano (GP) is an Italian hard cooked cheese characterized by a long ripening process and high protein and Ca contents. After in vitro static simulated gastrointestinal digestion, GP digest contained caseinophosphopeptides that were 6 to 24 amino acids in length, including tri-phosphorylated species incorporating the pSer-pSer-pSer-Glu-Glu cluster. Using rat ileum tissue, the digest was used to assess Ca absorption ex vivo, which showed significantly better results for the GP digest in comparison to the CaCO3 aqueous solution. An in vitro intestinal model based on Caco-2/HT-29 cell co-culture was able to mimic Ca absorption from GP digest, with Ca-rich water as a control. The metabolite-containing medium was then used to treat osteoblast-like SaOS-2 cells. As a consequence, metabolized GP digest significantly increased the number of osteoblasts, whereas the metabolized water did not exert this effect. Finally, the mice were fed diets containing GP or CaCO3 and pea isolate and the in vivo outcomes were assessed through fluorescent probe and computed tomography. Mice fed a diet containing GP showed a higher increase in bone remodeling and volume in comparison to those fed a control diet containing CaCO3 and pea isolate. Overall, the ex vivo, in vitro and in vivo experiments highlighted the effectiveness of GP in improving Ca absorption, osteoblast proliferation and bone remodeling and volume.
Keywords: calcium; caseinophosphopeptides; Ussing chambers; Caco-2/HT-29 cells; SaOS-2 cells; CD1 mice; bone remodeling calcium; caseinophosphopeptides; Ussing chambers; Caco-2/HT-29 cells; SaOS-2 cells; CD1 mice; bone remodeling
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MDPI and ACS Style

Martelli, C.; Ottobrini, L.; Ferraretto, A.; Bendinelli, P.; Cattaneo, S.; Masotti, F.; Stuknytė, M.; Dall’Asta, M.; Del Sole, A.; De Noni, I.; et al. Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese. Foods 2025, 14, 273. https://doi.org/10.3390/foods14020273

AMA Style

Martelli C, Ottobrini L, Ferraretto A, Bendinelli P, Cattaneo S, Masotti F, Stuknytė M, Dall’Asta M, Del Sole A, De Noni I, et al. Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese. Foods. 2025; 14(2):273. https://doi.org/10.3390/foods14020273

Chicago/Turabian Style

Martelli, Cristina, Luisa Ottobrini, Anita Ferraretto, Paola Bendinelli, Stefano Cattaneo, Fabio Masotti, Milda Stuknytė, Margherita Dall’Asta, Angelo Del Sole, Ivano De Noni, and et al. 2025. "Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese" Foods 14, no. 2: 273. https://doi.org/10.3390/foods14020273

APA Style

Martelli, C., Ottobrini, L., Ferraretto, A., Bendinelli, P., Cattaneo, S., Masotti, F., Stuknytė, M., Dall’Asta, M., Del Sole, A., De Noni, I., & Rossi, F. (2025). Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese. Foods, 14(2), 273. https://doi.org/10.3390/foods14020273

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