Effect of Hydrogen Oxide-Induced Oxidative Stress on Bone Formation in the Early Embryonic Development Stage of Chicken

Abstract

The current study aimed to monitor the impact of H₂O₂-induced oxidative stress on avian bone formation during the early stage of embryonic development. Fertilized Cobb broiler eggs were divided into five treatment groups and micro-injected with varying concentrations of H₂O₂, i.e., control (PBS; 0 nM), 10 nM, 30 nM, 100 nM, and 300 nM, on embryonic day 3, with continued incubation thereafter. The treatment concentrations were selected based on the level of lipid peroxidation and the survival rate of embryo. Embryos were collected at 6 h, 24 h, 48 h, and 72 h post-injection. The mRNA expression levels of apoptotic markers, antioxidant enzymes, and early bone formation gene markers were measured. The results showed that the microinjection of H₂O₂ altered the expression pattern of antioxidant enzymes' mRNA during early embryogenesis and decreased the expression of <i>COL1A2</i> and <i>COL2A1</i> at 6 h and 24 h post-injection. Decreased expression of <i>BMP, BGLAP</i>, and <i>RUNX2</i> was observed 48 h post-injection. Additionally, a shorter embryo length was observed in the 100 nM and 300 nM H₂O₂ treatment groups 72 h post-injection. In conclusion, H₂O₂-induced oxidative stress suppressed the expression of bone formation gene markers, with chronic effects on avian embryonic development.