Objective This study aims to evaluate the applicability of the ultrasound roughness index (URI) for quantitative assessment of cartilage quality ex vivo (post-mortem), after 6 months of in vivo articulation with a Focal Knee Resurfacing Implant (FKRI). Design Goats received a metal FKRI ( n = 8) or sham surgery ( n = 8) in the medial femoral condyles. After 6 months animals were sacrificed, tibial plateaus were stained with Indian ink, and macroscopic scoring of the plateaus was performed based on the ink staining. The URI was calculated from high-frequency ultrasound images at several sections, covering both areas that articulated with the implant and non-articulating areas. Cartilage quality at the most damaged medial location was evaluated with a Modified Mankin Score (MMS). Results The URI was significantly higher in the FKRI-articulating than in the sham plateaus at medial articulating sections, but not at sections that were not in direct contact with the implant, for example, under the meniscus. The mean macroscopic score and MMS were significantly higher in the FKRI-articulating group than in the sham group ([Formula: see text], [Formula: see text], respectively). Correlation coefficients between URI and macroscopic score were significant in medial areas that articulated with the implant. A significant correlation between URI and MMS was found at the most damaged medial location ([Formula: see text]). Conclusions This study demonstrates the potential of URI to evaluate cartilage roughness and altered surface morphology after in vivo articulation with a metal FKRI, rendering it a promising future tool for quantitative follow-up assessment of cartilage quality.