This study addresses the limitations of current tonometry techniques by exploring vibroacoustic properties for estimating intraocular pressure (IOP), a key diagnostic parameter for monitoring glaucoma—a significant risk factor for vision loss. Utilizing vivo porcine eyeballs, we investigated the relationship between IOP and the nonlinear vibration transfer function ratio (NVTFR). Through applying varying vibration levels and analyzing responses with transfer function analysis and univariate regression, we identified a strong negative correlation between NVTFR and IOP, evidenced by a Pearson correlation coefficient of −0.8111 and significant results from generalized linear model (GLM) regression (p-value < 0.001). These findings indicate the potential of NVTFR as a vital indicator of IOP changes. Our study highlights the feasibility of using vibroacoustic properties, specifically NVTFR, to measure IOP. While further refinement is necessary for in vivo application, this approach opens new possibilities for non-invasive and patient-friendly IOP monitoring, potentially enhancing ophthalmology diagnostic techniques and providing a foundation for future research and development in this critical area.