Activation of macrophages via toll‐like receptors (TLRs) is important for inflammation and host defense against pathogens. Recent data suggest that non‐pathogenic molecules released by trauma also can trigger inflammation via TLR2 and TLR4. Here, we tested whether TLRs are regulated after sterile spinal cord injury (SCI) and examined their effects on functional and anatomical recovery. We show that mRNA for TLR1, 2, 4, 5, and 7 are increased after SCI as are molecules associated with TLR signaling (e.g. MyD88, NFκB). The significance of in vivo TLR2 and TLR4 signaling was evident in SCI TLR4 mutant (C3H/HeJ) and TLR2 knockout (TLR2^−/−) mice. In C3H/HeJ mice, sustained locomotor deficits were observed relative to SCI wild‐type control mice and were associated with increased demyelination, astrogliosis, and macrophage activation. These changes were preceded by reduced intraspinal expression of interleukin‐1β mRNA. In TLR2^−/− mice, locomotor recovery also was impaired relative to SCI wild‐type controls and novel patterns of myelin pathology existed within ventromedial white matter – an area important for overground locomotion. Together, these data suggest that in the absence of pathogens, TLR2 and TLR4 are important for coordinating post‐injury sequelae and perhaps in regulating inflammation and gliosis after SCI.