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Updated: May 22, 2026

Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications
Published on: April 30, 2014
Evaluation of sterilization methods following contamination of hamstring autograft during anterior cruciate ligament
Matthew J Plante1, Xinning Li, Gail Scully
1Division of Sports Medicine, University of Massachusetts Medical Center, 55 Lake Avenue North, Worcester, MA, USA.
Purpose:
Inadvertent contamination of the hamstring autograft during ACL reconstruction is infrequent, but can result in significant complications. The purpose of this study is to evaluate bacterial contamination of hamstring autografts dropped onto the operating room floor and methods of graft decontamination.
Methods:
Hamstring tendons were harvested from patients. Excess tendon not used in the ACL procedure was divided into 6 segments. Segments were assigned to 6 groups (A through F, N = 30 in each group): group A: uncontaminated graft immediately postharvest (control), group B: graft dropped onto the floor (5 s), group C: graft dropped onto the floor (15 s). grafts in groups D to F were dropped onto floor for 15 s then rinsed with saline (group D), bacitracin solution (group E) or chlorhexidine 4 % solution (group F) for 3 min. All grafts were sent to the microbiology laboratory for anaerobic and aerobic cultures.
Results:
Cultures were positive in 23 % of graft segments from group A (7/30), 33 % of grafts from group B (10/30), 23 % from group C (7/30), 30 % from group D (9/30) and 3 % from both group E (1/30) and group F (1/30). Sixteen unique organisms were identified, with Staphylococcus aureus as the most common isolate. Grafts rinsed in either bacitracin solution or 4 % chlorhexidine solutions were significantly less likely to be culture positive when compared to control graft segments (p < 0.05). However, there was no significant difference between uncontaminated grafts retrieved in <5 versus 15 s from the floor.
Conclusion:
This study supports the practice of decontaminating a dropped ACL hamstring autograft using either 4 % chlorhexidine or bacitracin solution. Specimens should be retrieved sterilely and washed for at least 3 min. This study also demonstrates no advantage in retrieval time of less than 5 s as compared to 15 s for uncontaminated graft. Hamstring harvest in ACL reconstruction may result in positive cultures, thus routine soaking of the hamstring autograft in either bacitracin or 4 % chlorhexidine solution is recommended. In addition, dropped hamstring autograft can be effectively sterilized with bacitracin or 4 % chlorhexidine solution.
Level Of Evidence:
II.
Insights
Dropped hamstring autografts for ACL reconstruction can be effectively decontaminated using bacitracin or 4% chlorhexidine solutions. Routine soaking of the graft in these solutions is recommended to prevent complications from bacterial contamination.
Area of Science:
- Orthopedic Surgery
- Microbiology
- Biomedical Engineering
Background:
- Hamstring autografts are commonly used for anterior cruciate ligament (ACL) reconstruction.
- Inadvertent contamination of these grafts during surgery can lead to significant complications.
- Evaluating methods to decontaminate potentially contaminated grafts is crucial for patient safety.
Purpose of the Study:
- To assess bacterial contamination levels of hamstring autografts dropped on the operating room floor.
- To evaluate the efficacy of different decontamination methods for these grafts.
Main Methods:
- Hamstring tendon segments were harvested and divided into groups.
- Groups were subjected to different contamination durations (5s, 15s) and decontamination solutions (saline, bacitracin, 4% chlorhexidine).
- All graft segments underwent anaerobic and aerobic cultures to detect bacterial contamination.
Main Results:
- Positive cultures were observed in 23-33% of control and dropped grafts.
- Grafts decontaminated with bacitracin or 4% chlorhexidine showed significantly lower positive culture rates (3%) compared to controls (23%).
- No significant difference in contamination was found between grafts dropped for 5s versus 15s.
Conclusions:
- Bacitracin solution and 4% chlorhexidine effectively decontaminate dropped hamstring autografts.
- A minimum 3-minute washing period is recommended for decontamination.
- Routine soaking of hamstring autografts in these solutions is advised to minimize infection risk in ACL reconstruction.