Tibial subchondral bone plays an important role in knee osteoarthritis (OA). Microarchitectural characterization of subchondral bone plate (SBP), underlying subchondral trabecular bone (STB) and relationships between these compartments, however, is limited. The aim of this study was to characterize the spatial distribution of SBP thickness, SBP porosity and STB microarchitecture, and relationships among them, in OA tibiae of varying joint alignment. Twenty-five tibial plateaus from end-stage knee-OA patients, with varus (n = 17) or non-varus (n = 8) alignment were micro-CT scanned (17 μm/voxel). SBP and STB microarchitecture was quantified via a systematic mapping in 22 volumes of interest per knee (11 medial, 11 lateral). Significant within-condylar and between-condylar (medial vs. lateral) differences (p < 0.05) were found. In varus, STB bone volume fraction (BV/TV) was consistently high throughout the medial condyle, whereas in non-varus, medially, it was more heterogeneously distributed. Regions of high SBP thickness were co-located with regions of high STB BV/TV underneath. In varus, BV/TV was significantly higher medially than laterally, however, not so in non-varus. Moreover, region-specific significant associations between the SBP thickness and SBP porosity and the underlying STB microarchitecture were detected, which in general were not captured when considering the values averaged for each condyle. As subchondral bone changes reflect responses to local mechanical and biochemical factors within the joint, our results suggest that joint alignment influences both the medial-to-lateral and the within-condyle distribution of force across the tibia, generating corresponding local bony responses (adaptation) of both the subchondral bone plate and underlying subchondral trabecular bone microarchitecture.