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The study aimed to evaluate the potential of canola protein hydrolysates (CPH) on osteoclastogenesis using a pre\u2010osteoblast cell MC3T3\u2010E1. Twenty\u2010two CPHs were prepared by 12 different proteases, either individually or in combination. Three CPHs, prepared by trypsin (CPH\u2010T), Protex 6L (CPH\u2010P), and the combination of Protex 6L and thermoase (CPH\u2010PT) showed promising activity in promoting in vitro bone formation. CPH\u2010T and CPH\u2010PT improved cell proliferation at a concentration of 10 ug\/ml, while all three hydrolysates exhibited cytotoxicity at 1000 ug\/ml. All three hydrolysates promoted the level of runt\u2010related transcription factor 2 (RUNX2) and type I collagen, and mineralization in osteoblast cells, in a dose\u2010dependent manner. Additionally, these three hydrolysates elevated the osteoprotegerin (OPG) level and reduced the level of receptor activator of nuclear factor kappa\u2010B ligand (RANKL). This study indicated the activity of CPHs in the promotion of bone formation and prevention of osteoclastogenesis, suggesting the potential of CPHs as a promising functional food ingredient against osteoporosis.<\/jats:p>","DOI":"10.1111\/1750-3841.70210","type":"journal-article","created":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T06:49:45Z","timestamp":1745304585000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Canola Protein Hydrolysates Show Osteogenic Activity in MC3T3\u2010E1 Cells"],"prefix":"10.1111","volume":"90","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-0234-9654","authenticated-orcid":false,"given":"Da","family":"Shi","sequence":"first","affiliation":[{"name":"Department of Agricultural, Food and Nutritional Science, Faculty of Agricultural, Life and Environmental Sciences University of Alberta  Alberta 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