Kynurenine’s Effect on the Different Stages of Osteoblastic Differentiation

Abstract

This study focuses on the dynamics of age-related changes in osteoblasts’ matrix production, with a specific focus on the involvement of the aryl hydrocarbon receptor (AhR). The AhR is activated by kynurenine, a metabolite of the amino acid tryptophan that tends to increase with age. Kynurenine has been known to impair osteoblasts’ bone formation ability, however, the point at which this occurs is unknown. The study utilizes a 21-day differentiation process of mesenchymal stem cells. “Window experiments” that vary the intervals of kynurenine treatment to explore its impact at different stages of osteoblast development were utilized. The experimental setup involves four distinct conditions: a control group with a vehicle, a group treated with kynurenine, a group with an AhR antagonist (BAY), and a group treated with both kynurenine and BAY. The AhR antagonist, specifically BAY2416964, serves to investigate whether any observed effects of kynurenine are mediated through AhR signaling. Results, analyzed through alizarin red staining, demonstrate a notable negative impact of kynurenine on matrix production, particularly in the middle stages of osteoblast differentiation. The study further employs crystal violet staining to confirm the presence of cells in the experimental wells, providing assurance of the reliability of the alizarin staining procedure. Importantly, this staining procedure also suggests that the effects of kynurenine on mineralized matrix production are largely independent of any potential impacts on cell viability.