Tricuspid regurgitation (TR) is present in trace amounts or more in 82-86% of the population and is greater than mild in 14% of the population. In severe cases, it can contribute to right heart failure and adversely affect mitral valve repair durability. One major cause of TR is the dilation of the tricuspid annulus, which alters the geometry of the annulus from a saddle-shape to a more planar profile. Another cause of TR is the displacement of the papillary muscles (PMs), which results from right ventricular dilation. The objective of this study was to identify the effect of a saddle-shaped annulus on native tricuspid leaflet stretch mechanics and TR. In addition, the effects of geometric alterations, including annular dilatation and PM displacement, on leaflet stretch was investigated. Fresh porcine tricuspid valves (TVs) (n = 8) were excised and sutured to an adjustable three-dimensional annulus plate (allowing for dilatation and saddle-shape) and three PM attachment rods. The valve was then placed in the in vitro Georgia Tech right heart simulator. Dual-camera photogrammetry, was used to quantify the stretch ratio experienced by the valve leaflets at peak systole for the following conditions: physiologically normal, 100% annular dilatation, displaced PMs, and a combination of annular dilatation and PM displacement. In addition, a saddle and flat annulus were implemented for each of the four conditions. PM displacement was simulated by displacing all PMs by 10 mm in all directions (laterally, apically, posteriorly/anteriorly). The physiologically normal condition-normal annulus area, saddle-shaped annulus with PMs in a normal position, was used as a control. The results showed that the posterior leaflet exhibited significantly (p ≤ 0.05) higher major and areal stretch ratios as compared to the anterior leaflet at peak systole for all conditions tested. No significant difference was seen in stretch when a flat annulus was compared to saddle for the anterior or posterior leaflet for normal or disease conditions. Investigation of the impact of disease found a significant increase (p ≤ 0.10) in stretch in the posterior leaflet with a combination of annular dilatation and PM displacement (2.01 ± 0.68) as compared to the normal condition with a saddle annulus (1.43 ± 0.20). In addition displacement of the PMs resulted in a significant (p ≤ 0.01) reduction in TR, although the actual volume reduced was minimal (1.2 mL). Stretch values were measured for the anterior and posterior leaflet under both physiologic and pathologic conditions for the first time. Further, these results provide an understanding of the effects of geometric parameters on valve mechanics and function, which may lead to improved TV repairs.