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The autograft preparation process for anterior cruciate ligament reconstruction has a potential for graft contamination. The purpose of this study was to evaluate the possibility of contamination of the bone-patellar tendon-bone and hamstring tendon autograft during preparation for anterior cruciate ligament reconstruction.A primary isolated reconstruction of the anterior cruciate ligament with use of bone-patellar tendon-bone autograft (thirty patients) and hamstring tendon autograft (thirty patients) was performed in a prospective, consecutive series of patients. Three tissue samples were obtained for culture from each graft at different time-intervals during the graft preparation. In addition, the erythrocyte sedimentation rate and the C-reactive protein level were evaluated preoperatively and on the third, seventh, and twentieth postoperative days, and the clinical course of all patients was monitored.The time needed for graft preparation was significantly longer for hamstring autografts (nineteen minutes) than for bone-patellar tendon-bone autografts (ten minutes) (p = 0.032). In the hamstring group, cultures of graft tissue from four patients (13%) were positive for bacteria. In the bone-patellar tendon-bone group, cultures of graft tissue from three patients (10%) were positive for bacteria; the difference between groups was not significant (p = 0.923). No patient had development of a postoperative infection. There were no differences between patients with a contaminated graft and those with an uncontaminated graft with regard to postoperative changes in the erythrocyte sedimentation rate or the C-reactive protein level at all time-intervals.A high rate (12%) of autograft contamination can be expected during autograft preparation for anterior cruciate ligament reconstruction. The contamination rate is almost equal for both bone-patellar tendon-bone and hamstring tendon autografts. We could not identify an association between contaminated grafts implanted in the knee and postoperative inflammatory markers such as the erythrocyte sedimentation rate and the C-reactive protein level.Therapeutic Level II. See Instructions to Authors for a complete description of levels of evidence.