Nicotine is a chemical compound widely used for leisure, most commonly through cigarette smoking, but also distributed in the form of nicotine patches, gums, and e-cigarettes for smoking cessation. Its impact on bone health, however, remains controversial and not fully understood. Clinical studies have reported that smokers exhibit lower bone mineral density and increased risk of osteoporosis and fracture, but it remains unclear whether these outcomes arise from nicotine itself or from other components of tobacco smoke. In vitro studies have reported both stimulatory and inhibitory effects of nicotine on osteoblast proliferation, differentiation, and mineralization, depending on concentration and exposure duration. These conflicting findings highlight the need for controlled experimental models to explain the concentration-dependent and receptor-mediated effects of nicotine on bone cells. This project aims to elucidate the role of nicotine in bone metabolism using DBP-based bone organoids that recapitulate key structural and functional features of native bone tissue. By leveraging this three-dimensional organoid platform, this experiment will show how nicotine exposure modulates osteoblast activity, mineral deposition, osteoclast differentiation, and overall bone remodeling dynamics. This experiment will focus on the relationship between nicotine and nicotinic acetylcholine receptors (nAChRs) to understand the exact signaling mechanism by which nicotine affects bone-regulating cells and processes. Through this approach, the study will provide mechanistic insight into nicotine’s role in skeletal pathology and inform public health considerations for populations exposed to nicotine through both recreational and therapeutic delivery systems.