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Updated: May 14, 2025

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Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications
Published on: April 30, 2014
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Optimizing Vancomycin Soaking Protocols for Anterior Cruciate Ligament Reconstruction.
Wachiraphan Parinyakhup1, Piya Chavalparit1, Dennapa Saeloh Sotthibandhu2
1Department of Orthopedics, Faculty of Medicine, Prince of Songkla University, Hat Yai 90110, Songkhla, Thailand.
Antibiotics (Basel, Switzerland)
|April 29, 2025
Summary
Shortened vancomycin soaking of anterior cruciate ligament (ACL) autografts to 5 minutes effectively eradicates bacteria. Direct soaking at 5 mg/mL or gauze wrapping at 10 mg/mL eliminates pathogens without harming cells.
Area of Science:
- Orthopedic Surgery
- Infectious Disease Prevention
- Biomaterial Science
Background:
- Current anterior cruciate ligament (ACL) autograft guidelines recommend 20-minute vancomycin soaking.
- Clinical settings often limit soaking time to 5 minutes due to practical constraints.
- This study investigates vancomycin efficacy and safety within a 5-minute window.
Purpose of the Study:
- To assess bacterial eradication by vancomycin in ACL autografts within a 5-minute soak.
- To evaluate the cytotoxicity of different vancomycin concentrations and application methods.
- To determine optimal vancomycin protocols for preventing postoperative infections in ACL reconstruction.
Main Methods:
- Human semitendinosus tendons were inoculated with Staphylococcus aureus, Staphylococcus epidermidis (biofilm and non-biofilm), and Enterococcus faecalis.
- Treatment involved 5-minute direct soaking in 5 mg/mL vancomycin or gauze wrapping with 2.5, 5, or 10 mg/mL vancomycin.
- Bacterial elimination, cell viability (tenocytes, mesenchymal stem cells), and vancomycin release were quantified.
Main Results:
- Complete bacterial eradication was achieved with direct 5 mg/mL soaking and 10 mg/mL gauze wrapping.
- All tested vancomycin concentrations maintained cell viability above 70%, showing no significant cytotoxicity.
- Vancomycin release was highest with direct soaking but remained below chondrocyte toxicity thresholds.
Conclusions:
- A 5-minute direct vancomycin soak (5 mg/mL) or gauze wrap (10 mg/mL) effectively eliminates bacterial contamination on ACL autografts.
- These shortened protocols are safe, preserving tenocyte and mesenchymal stem cell viability.
- This research offers a practical, effective alternative for reducing infection risk in ACL reconstruction.
Keywords:
anterior cruciate ligament reconstructionbacterial eradicationcytotoxicityinfection preventionvancomycin soaking
