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Updated: Feb 21, 2026

Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications
Published on: April 30, 2014
Contamination occurs during ACL graft harvesting and manipulation, but it can be easily eradicated
Daniel Pérez-Prieto1, María E Portillo2, Raúl Torres-Claramunt3
1Department of Orthopaedic Surgery and Traumatology, Hospital del Mar, Universitat Autònoma de Barcelona (UAB), Passeig Marítim, 25, Barcelona, Spain. dperezprieto@parcdesalutmar.cat.
Purpose:
Why anterior cruciate ligament (ACL) autograft soaking in a 5 mg/ml vancomycin solution decreases the rate of infection has not been well-explained. One hypothesis is that grafts can be contaminated during harvesting and vancomycin eradicates the bacteria. The purpose of the present study is to assess how the vancomycin solution acts against ACL graft contamination during graft harvesting and preparation.
Methods:
The study was carried out in three university hospitals over a period of 6 months. After sample size calculation, 50 patients were included in the study. Three samples were taken from each ACL graft. Sample 1 was obtained immediately after graft harvesting. After graft manipulation and preparation, the remaining tissue was divided into two parts. The raw sample was denominated sample 2 and sample 3 consisted of the rest of the remaining tissue that had been soaked in the vancomycin solution. All the cultures were incubated at 37 °C with 5% CO2 in agar plates for 7 days (aerobically) or 14 days (anaerobically) and inspected daily for microbial growth. Any bacterial growth and the number of colony forming units were reported.
Results:
In seven cases (14%), either sample 1 or sample 2 was positive. In five of the cases (10%), only the sample after graft preparation was positive (sample 2). In two cases (4%), sample 1 and sample 2 were positive for the same bacteria. Isolated microorganisms corresponded to coagulase-negative staphylococci (CNS) and Propionibacterium acnes. No bacterial growth was observed in sample 3 (p < 0.001). Thus, none of those seven positive cases (0%) were positive after vancomycin soaking (p < 0.001).
Conclusion:
In the series, ACL graft harvesting and manipulation leads to bacterial contamination in 14% of the cases. This contamination is fully eradicated after soaking in the vancomycin solution in this series.
Level Of Evidence:
Level II.
Insights
Vancomycin soaking effectively eliminates bacterial contamination in anterior cruciate ligament (ACL) grafts, with 14% of grafts showing contamination post-harvesting. This study confirms vancomycin
Area of Science:
- Orthopedic Surgery
- Microbiology
- Infectious Disease Prevention
Background:
- Anterior cruciate ligament (ACL) reconstruction is a common orthopedic procedure.
- Surgical site infections following ACL reconstruction can lead to poor outcomes.
- Vancomycin soaking of ACL autografts is hypothesized to reduce infection rates by eradicating bacteria.
Purpose of the Study:
- To investigate the efficacy of vancomycin solution in eradicating bacterial contamination of ACL grafts.
- To assess the rate of ACL graft contamination during harvesting and preparation.
Main Methods:
- A prospective study involving 50 patients undergoing ACL reconstruction across three university hospitals.
- Bacterial samples were collected from ACL grafts at three stages: immediately post-harvesting (Sample 1), after preparation (Sample 2), and after vancomycin soaking (Sample 3).
- Cultures were incubated for 7 (aerobic) or 14 (anaerobic) days, with daily monitoring for microbial growth and colony-forming units.
Main Results:
- Bacterial contamination was detected in 14% of ACL grafts (7 out of 50 patients) after harvesting and preparation (Samples 1 and/or 2).
- Commonly isolated microorganisms included coagulase-negative staphylococci (CNS) and Propionibacterium acnes.
- No bacterial growth was observed in any graft samples after soaking in vancomycin solution (Sample 3), indicating complete eradication (p < 0.001).
Conclusions:
- ACL graft harvesting and preparation procedures can lead to significant bacterial contamination in 14% of cases.
- Soaking ACL grafts in a 5 mg/ml vancomycin solution effectively eradicates all detected bacterial contamination.
- Vancomycin soaking represents a crucial step in preventing graft-related infections during ACL reconstruction.
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