Background: Tympanosclerosis is the calcification of connective tissue in the middle ear, particularly the tympanic membrane, which can arise following middle ear infection. Inflammatory reactions are induced and maintained by tumour necrosis factor (TNF) in a variety of illnesses, including chronic otitis media with effusion (COME). The goal of this study was to determine the plasma level of Tumer necrosis factor (TNF) gene expression in individuals who had tympanosclerosis as a result of ventilation tube insertion. Methods: This study comprised sixty patients who were separated into two groups: 60 patients were assigned to Group A based on inclusion and exclusion criteria, whereas 20 volunteers were assigned to Group B. Patients were given a medical history, a thorough physical examination, and a TNF- gene expression test in the lab (qRT-PCR). Results: There was a significant difference in TNF-gene expression across groups, with the mean TNF in the cases group (3.87) being significantly greater than the control group (1), p0.001. TNF levels were statistically different depending on the number of tubes inserted (p=0.005) and the time of secretory OM (p=0.042). There was no significant difference in TNF levels when it came to medication usage or acute OM episodes. Conclusion: TNF- gene profile expression was statistically significant higher in selected middle-ear structures with tympanosclerosis than in the control group.