There is a need to understand how anthropogenic influences affect urban and periurban forest diversity at the regional scale. This study aims to compare urban and periurban tree composition along a geographic gradient, and test hypotheses about species composition and ecological homogeneity. We paired urban forest (UF) data from eight cities across the southeastern US with periurban forest (PF) data from the USDA Forest Service Forest Inventory and Analysis program. We found that tree diversity, as well as both observed and estimated species richness values were greater in UF versus PF. Community size structure analysis also indicated a greater proportion of large trees and greater numbers of non-native, invasive, and unclassified tree species in the UF versus the PF, regardless of location. Both forest type and ecological province had a significant effect on community species composition, with forests closer together in space being more similar to each other than those more distant. While land use change and management has been associated with ecological homogenization in human dominated landscapes, we found that species composition was more dissimilar along latitudinal lines than compared to between forest types, refuting this hypothesis, at least in terms of tree diversity.