The study aimed to use beta-glucan extracted from barley to prepare eco-friendly silver nanoparticles (AgNPs) using green synthesis methods. The barley variety "Abah 99" was used, with an extraction yield of 78% and a concentration of 4.71%. Field effect scanning electron microscopy (FESEM) was used, revealing an irregular spherical shape with sizes ranging from 13.829 to 56.233 nm. Energy-dispersive X-ray (EDX) analysis showed the presence of Ag atoms at specific ratios in the sample. Zeta potential analysis confirmed the presence of negative charges on the green-synthesized particles (-18.7 mV). X-ray diffraction (XRD) analysis showed prominent peaks at 38.19, 44.37, 64.56, and 77.47, corresponding to the (111), (200), (220), and (311) planes, respectively. Furthermore, an MTT assay was conducted on HEK-293 cells (a type of human embryonic kidney cell) exposed to green-synthesized beta-glucan-integrated silver nanoparticles. Cell viability decreased in a dose-dependent manner following AgNP treatment. However, treatment with Ag-beta glucan green synthesis resulted in very high cell viability (approximately 100%) compared to the chemically synthesized silver nanoparticles at concentrations of 3.125, 6.25, and 12.5 µg/mL. The study demonstrated that cell viability decreased with higher concentrations, with the chemically synthesized  AgNPs showing a notable reduction in cell viability, especially at concentrations of 25, 50, and 100 µg/mL. In contrast, the green synthesis (AgNPs-beta glucan) showed decreased cell viability only at 50 and 100 µg/mL, while the extract group maintained better cell survival at a concentration of 100 µg/mL.