ORIGINAL RESEARCH article
Front. Nutr.
Sec. Nutrition and Food Science Technology
Free amino acid–rich egg yolk protein hydrolysate promotes osteogenesis of MC3T3-E1 cells association with β-catenin nuclear translocation
Provisionally accepted
- 1North Sichuan Medical College, Nanchong, China
- 2Hallym University, Chuncheon-si, Republic of Korea
- 3MEDIENCE Co Ltd, Chuncheon-si, Republic of Korea
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Background: Promoting osteogenesis is a key approach to preventing and improving bone metabolic diseases. Objective: This study investigated the impact of free amino acid-rich egg yolk protein hydrolysate (Y-PEP) on MC3T3-E1 osteoblast differentiation and mineralization. Methods: Free amino acids in Y-PEP were quantified using a high-speed amino acid analyzer. MC3T3-E1 cells were osteogenically induced with Y-PEP (5–100 μg/mL), and cell viability (72 h), alkaline phosphatase activity and collagen synthesis (day 9), as well as calcium deposition and osteocalcin production (day 18) were assessed. Moreover, runt-related transcription factor 2/osterix mRNA (qRT-PCR) and total/cytosolic/nuclear β-catenin and β-catenin phosphorylation (western blot) were measured. Results: Twenty-one amino acids, including leucine, lysine, arginine, glutamic acid, and valine, were identified and quantified in Y-PEP by comparing retention times and peak areas with amino acid mixture standard solutions. Y-PEP concentrations below 100 μg/mL had no impact on MC3T3-E1 osteoblast viability. Y-PEP enhanced osteoblast differentiation markers in a dose-dependent manner at concentrations from 5–100 μg/mL and promoted mineralization markers in mature osteoblasts dose-dependently at 25–50 μg/mL. The pro-osteogenic effect of Y-PEP may involve increasing total cellular β-catenin levels and promoting β-catenin nuclear translocation rate to upregulate transcription of osteogenesis-associated genes. The osteogenic activity of Y-PEP may result from the synergistic effects among signal transduction, metabolism, and mineral handling driven by its complex amino acid composition. Conclusion: Y-PEP can promote osteogenesis in MC3T3-E1 cells and has the potential to serve as a functional food ingredient for preventing or improving metabolic bone diseases.
Keywords: Egg yolk protein hydrolysate, Free amino acid, MC3T3-E1 osteoblasts, Osteogenesis, β-catenin
Received: 24 Dec 2025; Accepted: 05 Feb 2026.
Copyright: © 2026 Zheng, Park, Hwang, Kim, Park and Kang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Il-Jun Kang
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