Protein bioactive complexes promote osteogenesis under microgravity environment.

The space microgravity environment and cosmic radiation pose a significant threat to musculoskeletal health, particularly bone mass. However, the critical mechanism underlying space-induced bone loss and its relation to cellular oxidative stress remains unclear. Currently used bone-loss-reversing drugs face limitations like poor efficacy and metabolic defects. Herein, we revealed that simulated microgravity (SMG) induces reactive oxygen species (ROS), negatively impacting osteoblasts, causing cytoskeletal damage, and downregulating osteogenic genes. To combat this, we designed protein-zein nanocages loaded with a chimeric non-enzymatic cocktail (ZNAC) containing ascorbic acid, resveratrol, luteolin, coenzyme Q, and glutathione. These nanocages (~200 nm) demonstrated excellent stability, biocompatibility, and antioxidant properties compared to free drugs. We investigated the effects of ZNAC under SMG using two experimental models: MC3T3 pre-osteoblast/MG63 osteoblasts and regenerating zebrafish scales that represent compositional and physiological/pathophysiological analogy with mammalian system. ZNAC effectively reduced SMG-induced ROS, preserved cytoskeletal integrity, and enhanced alkaline phosphatase (ALP) activity along with the expression of osteogenic genes such as RUNX2 and Col1A1. In zebrafish scales, it increased osteogenic gene expression, calcification, and the calcium/phosphorus ratio, indicating enhanced scale regeneration. These findings suggest that ZNAC is a promising candidate for bone regeneration, offering potential solutions for maintaining astronaut health during extended space missions.