Nephrolithiasis is one of the common renal diseases that is attributed to multiple factors. On the other hand, calcium oxalate is the most common urine lithiasis, and urinary calcium and oxalate oversaturation is vital to calcium oxalate stone formation. The percent study examined the possible role of Ethylene Glycol-Induced Nephrolithiasis on Calcium Oxalate Crystals and Oxidative Damage of Renal Cells in stone formation. The ethylene glycol was administered at doses of 0.4% (LD), 0.75% (MD), and 1.0% (HD) in drinking water for 28 days in Wistar rats to induce Nephrolithiasis. The blood and urine samples were collected on 14 and 28 days of treatment and analyzed for various biochemical parameters. Ethylene glycol causes a significant dose-dependent reduction in body weight and dose and time-dependent increases in urinary calcium, oxalate, phosphate, and total protein excretion levels and decreased urinary magnesium excretion in urine samples. Ethylene glycol causes dose and time-dependent increases in calcium, phosphate, and total protein content and significantly reduces magnesium.