This work aimed to investigate the feasibility of using and mixing Toona ciliata, Eucalyptus grandis/urophylla and Pinus oocarpa woods in OSB production. Three one-species and four mixed-species combinations were compared. Layer structure varied by positioning different wood species in the surface and core of the panels. Phenol-formaldehyde (PF) adhesive was applied at 9% for all OSB panels. Three-layer mats with mass proportion of 25/50/25% for surface/core/surface were produced. Pressing time was 8 min, under a 4 MPa pressure and temperature of 180 ˚C. Physical and mechanical properties were evaluated and compared to EN (300) commercialization standard. Most OSB panels did not fully attain such requirements. OSB panels made with Eucalyptus grandis/urophylla and Pinus oocarpa woods have potential to be commercialized as OSB/1 and OSB/2 types, respectively. Among panels made with T. ciliata wood, those produced with this species in the surface and Eucalyptus grandis/urophylla wood in the core met the requirements established for OSB/1 commercialization. Eucalyptus wood has great potential to substitute Pinus wood in OSB production. The utilization of T. ciliata wood for OSB production decreased mechanical performance, but remarkably enhanced water resistance properties.