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 COL1A2 and COL2A1 at 6 h and 24 h post-injection. Decreased expression of BMP, BGLAP, and RUNX2 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.

Graded levels of Eimeria challenge altered the microstructural architecture and reduced the cortical bone growth of femur of Hy-Line W-36 pullets at early stage of growth (0–6 wk of age)

“…The oxidative stress markers, including total antioxidant capacity, superoxide dismutase, malondialdehyde, glutathione, and glutathione reductase, were significantly different in challenged birds from those in control, indicating the challenged pullets suffered from oxidative stress ( Sharma et al, 2022a ). It has been reported that ROS inhibits the differentiation of osteoblast cells and promotes osteoblast apoptosis, which decreases the rate of bone formation ( Domazetovic et al, 2017 ; Tian et al, 2017 ; Tompkins et al, 2023a , b , c ). Recently, Tompkins et al (2023b , c ) reported that ROS induced by H 2 O 2 significantly inhibits osteogenic differentiation of mesenchymal stem cells isolated chicken compact bones and skeletal development of chick embryos, indicating that ROS production in the body is a key inhibitor for bone cell formation and bone tissue development.…”

“…It has been reported that ROS inhibits the differentiation of osteoblast cells and promotes osteoblast apoptosis, which decreases the rate of bone formation ( Domazetovic et al, 2017 ; Tian et al, 2017 ; Tompkins et al, 2023a , b , c ). Recently, Tompkins et al (2023b , c ) reported that ROS induced by H 2 O 2 significantly inhibits osteogenic differentiation of mesenchymal stem cells isolated chicken compact bones and skeletal development of chick embryos, indicating that ROS production in the body is a key inhibitor for bone cell formation and bone tissue development. In addition, ROS plays a crucial role in osteoclast differentiation by increasing the production of receptor activators of nuclear factor-kappa B ligand ( RANKL ).…”

“…There is enough evidence to support an adverse effect of coccidiosis in broilers, such as reduced bone mineralization leading to greater porosity in the cortical bone, reduced bone mineral density, reduced bone strength, and increased trabecular bone resorption ( Oikeh et al, 2019b ; Sakkas et al, 2019 ; Shi et al, 2022 ; Tompkins et al, 2022 , 2023a ). Previously, the deleterious effect in bone mineralization following coccidiosis was related to malabsorption and nutrient deficiency ( Turk, 1973 , 1978 ); however, oxidative stress and inflammation might play a crucial role in bone growth ( Domazetovic et al, 2017 ; Tompkins et al, 2022 , 2023a , b , c ). Skeletal health is of utmost importance in laying hens, as their long bones in particular serve as a labile source of calcium for both egg production and providing mechanical support for their entire body.…”

Oligomeric proanthocyanidins ameliorates osteoclastogenesis through reducing OPG/RANKL ratio in chicken's embryos

“…In addition, antioxidant capacity is believed to be related to immunity and bone health of layer chickens. Tompkins et al ( Tompkins et al, 2023 ) found that hydrogen peroxide ( H 2 O 2 )-induced oxidative stress has an inhibitory effect on the expression of osteogenesis markers, such as COL1A2, collagen type II alpha 1 ( COL2A1 ), bone morphogenetic protein ( BMP ), OCN and Runx2, and mediates the activity of antioxidant enzymes of chicken embryos. Therefore, dietary supplementation with antioxidants can against the potential oxidative stress, and eventually contribute to the integrity of bone structure and metabolism in chickens.…”

The Functional Roles of Methionine and Arginine in Intestinal and Bone Health of Poultry: Review

“…A recent study investigating the impact of hydrogen peroxide-induced oxidative stress on chicken compact bone-derived MSCs revealed that oxidative stress significantly increased apoptosis and decreased osteogenic differentiation of the cells, and this was accompanied by a universal decrease in osteogenesis-related gene expression and a decline in vitro mineralization [ 161 ]. Likewise, another study found that oxidative stress suppressed the expression of osteogenesis-related genes in chicken embryos, consequently impairing proper bone development [ 162 ]. Both studies confirmed the negative effects of oxidative stress on chicken bone development.…”