The accelerating pace of urbanization and population growth has increasingly disrupted the relationship between the built environment and nature, contributing to a decline in individuals’ psychological and spiritual well-being. Religious buildings—particularly mosques—have the potential to serve as restorative environments that support spiritual reflection and emotional balance. Within this context, the integration of biophilic design principles into sacred architecture has emerged as a significant area of contemporary research. This study investigates how biophilic elements in mosque interiors influence individuals’ perceptions of spirituality and the restorative quality of the space. Mosque interior images were generated using artificial intelligence tools (Midjourney V6.1 and DALL·E 3), informed by biophilic design theory. Grounded in Attention Restoration Theory (ART), the study utilized a quantitative research framework to explore how biophilic elements influence spiritual and restorative perceptions in sacred architectural spaces. Data were collected from 390 individuals in Türkiye with prior worship experience in Republican-era mosques—structures marked by ideological and spatial transformations. Two instruments were used, the Perceived Restorativeness Scale (PRS) and the newly developed Biophilic Spiritual Perception Scale (BSPS), which demonstrated high internal consistency (Cronbach’s α = 0.981). Analyses included exploratory and confirmatory factor analyses, as well as parametric and non-parametric statistical tests. Findings suggest that biophilic design in mosque interiors positively influences both spiritual experience and perceived environmental restorativeness. These results support the view that biophilic design should not be seen merely as an esthetic or ecological approach but as a multidimensional strategy that enhances the emotional and spiritual quality of sacred spaces. Implications for future mosque design in Türkiye are discussed.