The purpose of this study is to compare the local microfracture effects of antegrade versus retrograde drilling of the tibial tunnel in ACL reconstruction. Arthroscopic ACL excision was performed on eight matched cadaveric knees. Arthroscopic guided tibial tunnel reaming was performed in either an antegrade (four) or retrograde (four) direction. A 3 x 3 cm section of proximal tibial surrounding the tibial aperture was removed with open dissection, and each section underwent micro-computed tomography analysis. Three musculoskeletal radiologists graded the specimens for bone aperture disruption and discrete fracture lines. Tibial aperture irregularity was seen in all four of the antegrade specimens (mean, Grade 1.5), and in none of the retrograde specimens. Discrete fracture lines were present in all four antegrade specimens (mean 10.13 mm depth; 8.95 mm length). No fracture lines were seen in the retrograde group. Retrograde drilling of the tibial tunnel in ACL reconstruction results in less microfracture trauma to the surrounding aperture bone. The use of retrograde drilling in ACL reconstruction may decrease synovialization of the graft-tissue interface when compared to antegrade drilling.