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

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 s...

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Main Authors: Cristina Martelli, Luisa Ottobrini, Anita Ferraretto, Paola Bendinelli, Stefano Cattaneo, Fabio Masotti, Milda Stuknytė, Margherita Dall’Asta, Angelo Del Sole, Ivano De Noni, Filippo Rossi
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Language:English
Published: MDPI AG 2025-01-01
Series:Foods
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Online Access:https://www.mdpi.com/2304-8158/14/2/273
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author Cristina Martelli
Luisa Ottobrini
Anita Ferraretto
Paola Bendinelli
Stefano Cattaneo
Fabio Masotti
Milda Stuknytė
Margherita Dall’Asta
Angelo Del Sole
Ivano De Noni
Filippo Rossi
author_facet Cristina Martelli
Luisa Ottobrini
Anita Ferraretto
Paola Bendinelli
Stefano Cattaneo
Fabio Masotti
Milda Stuknytė
Margherita Dall’Asta
Angelo Del Sole
Ivano De Noni
Filippo Rossi
author_sort Cristina Martelli
collection DOAJ
description 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 CaCO<sub>3</sub> 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 CaCO<sub>3</sub> 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 CaCO<sub>3</sub> 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.
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spelling doaj-art-aec44f5064a341488072bccad6badb692025-01-24T13:33:06ZengMDPI AGFoods2304-81582025-01-0114227310.3390/foods14020273Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano CheeseCristina Martelli0Luisa Ottobrini1Anita Ferraretto2Paola Bendinelli3Stefano Cattaneo4Fabio Masotti5Milda Stuknytė6Margherita Dall’Asta7Angelo Del Sole8Ivano De Noni9Filippo Rossi10Department of Pathophysiology and Transplantation, University of Milan, 20054 Milan, ItalyDepartment of Pathophysiology and Transplantation, University of Milan, 20054 Milan, ItalyDepartment of Biomedical Sciences for Health, University of Milan, 20133 Milan, ItalyDepartment of Biomedical Sciences for Health, University of Milan, 20133 Milan, ItalyDepartment of Food, Environmental and Nutritional Sciences, University of Milan, 20133 Milan, ItalyDepartment of Food, Environmental and Nutritional Sciences, University of Milan, 20133 Milan, ItalyUnitech COSPECT—COmprehensive Substances characterization via advanced sPECTtroscopy, University Technological Platform, University of Milan, 20133 Milan, ItalyDepartment of Animal Science, Food and Nutrition, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Health Sciences, University of Milan, 20146 Milan, ItalyDepartment of Food, Environmental and Nutritional Sciences, University of Milan, 20133 Milan, ItalyDepartment of Animal Science, Food and Nutrition, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyGrana 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 CaCO<sub>3</sub> 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 CaCO<sub>3</sub> 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 CaCO<sub>3</sub> 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.https://www.mdpi.com/2304-8158/14/2/273calciumcaseinophosphopeptidesUssing chambersCaco-2/HT-29 cellsSaOS-2 cellsCD1 mice
spellingShingle Cristina Martelli
Luisa Ottobrini
Anita Ferraretto
Paola Bendinelli
Stefano Cattaneo
Fabio Masotti
Milda Stuknytė
Margherita Dall’Asta
Angelo Del Sole
Ivano De Noni
Filippo Rossi
Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese
Foods
calcium
caseinophosphopeptides
Ussing chambers
Caco-2/HT-29 cells
SaOS-2 cells
CD1 mice
title Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese
title_full Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese
title_fullStr Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese
title_full_unstemmed Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese
title_short Ex Vivo, In Vitro and In Vivo Bone Health Properties of Grana Padano Cheese
title_sort ex vivo in vitro and in vivo bone health properties of grana padano cheese
topic calcium
caseinophosphopeptides
Ussing chambers
Caco-2/HT-29 cells
SaOS-2 cells
CD1 mice
url https://www.mdpi.com/2304-8158/14/2/273
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