Selenium (Se) plays a crucial role in modulating inflammation and oxidative stress within the human system. Biogenic selenium nanoparticles (SeNPs) synthesized by Lactobacillus casei (L. casei) exhibit anti-inflammatory and anti-oxidative properties, positioning them as a promising alternative to traditional supplements characterized by limited bioavailability. With this context in mind, this study investigates the impact of selenium and L. casei in ameliorating inflammation and oxidative stress using a cell line model. The study is centered on the biosynthesis of selenium nanoparticles (SeNPs) by L. casei 393 under anaerobic conditions using a solution of sodium selenite (Na2SeO3) in the bacterial culture medium. The generation of SeNPs ensued from the interaction of L. casei bacteria with selenium ions, a process characterized via transmission electron microscopy (TEM) to confirm the synthesis of SeNPs. To induce inflammation, the human colonic adenocarcinoma cell line, Caco-2 was subjected to interleukin-1 beta (IL-1β) at concentrations of 0.5 and 25 ng/ml. Subsequent analyses encompass the evaluation of SeNPs derived from L. casei, its supernatant, commercial selenium, and L. casei probiotic on Caco2 cell line. Finally, we assessed the inflammatory and oxidative stress markers. The assessment of inflammation involved the quantification of NF-κB and TGF-β gene expression levels, while oxidative stress was evaluated through the measurement of Nrf2, Keap1, NOX1, and SOD2 gene levels. L. casei successfully produced SeNPs, as confirmed by the color change in the culture medium and TEM analysis showing their uniform distribution within the bacteria. In the inflamed Caco-2 cell line, the NF-κB gene was upregulated, but treatment with L. casei-SeNPs and selenium increased TGF-β expression. Moreover, L. casei-SeNPs upregulated SOD2 and Nrf2 genes, while downregulating NOX1, Keap1, and NF-κB genes. These results demonstrated the potential of L. casei-SeNPs for reducing inflammation and managing oxidative stress in the Caco-2 cell line. The study underscores the ability of L. casei-SeNPs to reduce oxidative stress and inflammation in inflamed Caco-2 cell lines, emphasizing the effectiveness of L. casei as a source of selenium. These insights hold significant promise for the development of SeNPs derived from L. casei as potent anti-inflammatory and anti-cancer agents, paving the way for novel therapeutic applications in the field.