The development of scaffolding materials for clinical applications depends on a thorough understanding of the mechanics of tissue-material interaction. Mechanical failure or failure strength of materials has a key role in their performance particularly in load bearing applications. An ideal scaffolding material should have the physical and mechanical properties as close as possible to those of the tissue being replaced as the same time as supporting cell adhesion and growth while maintaining its shape. Regarding the synthetic polymers used for scaffold preparation, polyvinyl alcohol (PVA) is one of the first synthetic macromolecules tested to produce spongy matrices for tissue regeneration. PVA has broad applications including tissue mimicking, vascular cell culture, vascular implanting, heart valves, cartilage substitute, contact lenses, and corneal implants. There is however, a lack of sufficient data on the mechanical failure of PVA-based materials and this brings about limited application of PVA in tissue engineering applications. In this study, a biomechanical method is used to characterize the mechanical failure of fabricated PVA sponge (P-sponge) and compared with two commercially available sponges (CENEFOM and EYETEC) using a biaxial puncture testing machine. It is revealed that the failure stress of P-sponge is 9.91% and 3.85% higher than that of the CENEFOM and EYETEC sponges. It is hoped that the mechanical information obtained here can provide insight into biomechanical approaches that elucidate the potential causes of early rupture, and suggest future treatments for prevention of mechanical failure of PVA sponges used as scaffolding materials.