Osteoanabolic activity of whey-derived anti-oxidative (MHIRL and YVEEL) and angiotensin-converting enzyme inhibitory (YLLF, ALPMHIR, IPA and WLAHK) bioactive peptides

Exploring bone rebuilding anabolic agents has been gaining much attention due to their potential therapeutic effects in treating several bone disorders including osteoporosis. Whey protein has been reported to affect bone health osteoanabolically, in terms of proliferation and differentiation of primary osteoblast cells. This study investigates whether whey derived anti-oxidative (AO) (P1- MHIRL, P2- YVEEL) and angiotensin converting enzyme inhibitory (ACE inhibitory) (P3- YLLF, P4-ALPMHIR, P5-IPA, P6- WLAHK) bioactive peptides affect the proliferation and differentiation of primary osteoblast cells isolated from rat calvaria. The proliferation and osteogenic activity of osteoblast cells in presence of these peptides were determined by MTT assay, DNA quantification study, Alkaline phosphatase activity (ALP) and ALP staining, Alizarin red activity and staining, and secretory osteocalcin measurement. The expression of osteogenesis-related genes (COLI-α, ALP, OCN and RUNX2) were determined by real-time quantitative PCR (RT-PCR) analysis over a period of 21 days. The peptide treated osteoblasts showed a significant increase in viable cell density and proliferation in the order of P2 > P6 > P3 at optimised concentration. Furthermore, the osteoblastic differentiation markers in response to these peptides were found to be significantly up regulated in the order of P2 > P6 > P3 when compared to the controls. These results demonstrated that bioactive whey-derived AO and ACE inhibitory peptides can play a potential therapeutic role in osteoporosis by activating osteoblasts anabolically.