Plastics, especially polystyrene nanoplastic particles (PSNPs), are known for their durability and absorption properties, allowing them to interact with environmental pollutants such as di-n-butyl phthalate (DBP). Previous research has highlighted the potential of these particles as carriers for various pollutants, emphasizing the need to understand their environmental impact comprehensively. This study focuses on the subchronic exposure of male Swiss albino mice to PSNP and DBP, aiming to investigate their reproductive toxicity between these pollutants in mammalian models. The primary objective of this study is to examine the reproductive toxicity resulting from simultaneous exposure to PSNP and DBP in male Swiss albino mice. The study aims to analyze sperm parameters, measure antioxidant enzyme activity, and conduct histopathological and morphometric examinations of the testis. By investigating the individual and combined effects of PSNP and DBP, the study seeks to gain insights into their impact on the reproductive profile of male mice, emphasizing potential synergistic interactions between these environmental pollutants. Male Swiss albino mice were subjected to subchronic exposure (60days) of PSNP (0.2mg/m, 50nm size) and DBP (900mg/kg bw), both individually and in combination. Various parameters, including sperm parameters, antioxidant enzyme activity, histopathological changes, and morphometric characteristics of the testis, were evaluated. The Johnsen scoring system and histomorphometric parameters were employed for a comprehensive assessment of spermatogenesis and testicular structure. The study revealed non-lethal effects within the tested doses of PSNP and DBP alone and in combination, showing reductions in body weight gain and testis weight compared to the control. Individual exposures and the combination group exhibited adverse effects on sperm parameters, with the combination exposure demonstrating more severe outcomes. Structural abnormalities, including vascular congestion, Leydig cell hyperplasia, and the extensive congestion in tunica albuginea along with both ST and Leydig cell damage, were observed in the testis, underscoring the reproductive toxicity potential of PSNP and DBP. The Johnsen scoring system and histomorphometric parameters confirmed these findings, providing interconnected results aligning with observed structural abnormalities. The study concludes that simultaneous exposure to PSNP and DBP induces reproductive toxicity in male Swiss albino mice. The combination of these environmental pollutants leads to more severe disruptions in sperm parameters, testicular structure, and antioxidant defense mechanisms compared to individual exposures. The findings emphasize the importance of understanding the interactive mechanisms between different environmental pollutants and their collective impact on male reproductive health. The use of the Johnsen scoring system and histomorphometric parameters provides a comprehensive evaluation of spermatogenesis and testicular structure, contributing valuable insights to the field of environmental toxicology.