Michelle Xiao1, Erika A Leonardi1, Orr Sharpe2, Seth L Sherman1, Marc R Safran1, William H Robinson2,3, Geoffrey D Abrams1. 1. Department of Orthopaedic Surgery, Stanford University School of Medicine, Palo Alto, California, USA. 2. Veterans Administration Palo Alto Health Care System, Palo Alto, California, USA. 3. Department of Medicine, Division of Rheumatology, Stanford University School of Medicine, Palo Alto, California, USA.
Abstract
BACKGROUND: Surgical site infections (SSIs) after anterior cruciate ligament (ACL) reconstruction procedures are an unfortunate complication. Soaking grafts in vancomycin before implantation has been reported to reduce the incidence of postoperative SSI after ACL reconstruction. There is potential for vancomycin to compromise graft integrity because of tenocyte toxicity. PURPOSE: To examine the in vitro toxicity of varying doses of vancomycin on human tenocytes. STUDY DESIGN: Controlled laboratory study. METHODS: Human patellar tenocytes were isolated and expanded in vitro. Tenocytes in culture were exposed to vancomycin at 5 different concentrations (400, 1600, 3200, 6400, and 12,800 μg/mL) and 3 time intervals (2, 6, and 24 hours). The control for all series was tenocyte exposure to only culture medium for each time interval. After treatment, a 10% Cell Counting Kit-8 solution in cellular growth medium was applied to the cells to examine cytotoxicity. A live/dead assay was used to assess tenocyte viability through fluorescence microscopy and flow cytometry. Results were analyzed statistically using multivariable logistic regression models with Tukey honest significant difference post hoc tests. RESULTS: Vancomycin did not cause significant changes in tenocyte viability after 2 and 6 hours of incubation at any concentration between 0 and 12,800 µg/mL. Incubation with vancomycin for 24 hours led to a significant decrease in cell viability at higher concentrations. CONCLUSION: Tenocytes derived from human patellar tendons exposed to relatively high concentrations of vancomycin for short periods of time do not demonstrate significant cell death and toxicity. CLINICAL RELEVANCE: Exposing tendons to vancomycin for a short period of time, such as before ACL reconstruction, is not likely to cause tenocyte toxicity because of vancomycin administration.
BACKGROUND: Surgical site infections (SSIs) after anterior cruciate ligament (ACL) reconstruction procedures are an unfortunate complication. Soaking grafts in vancomycin before implantation has been reported to reduce the incidence of postoperative SSI after ACL reconstruction. There is potential for vancomycin to compromise graft integrity because of tenocyte toxicity. PURPOSE: To examine the in vitro toxicity of varying doses of vancomycin on human tenocytes. STUDY DESIGN: Controlled laboratory study. METHODS:Human patellar tenocytes were isolated and expanded in vitro. Tenocytes in culture were exposed to vancomycin at 5 different concentrations (400, 1600, 3200, 6400, and 12,800 μg/mL) and 3 time intervals (2, 6, and 24 hours). The control for all series was tenocyte exposure to only culture medium for each time interval. After treatment, a 10% Cell Counting Kit-8 solution in cellular growth medium was applied to the cells to examine cytotoxicity. A live/dead assay was used to assess tenocyte viability through fluorescence microscopy and flow cytometry. Results were analyzed statistically using multivariable logistic regression models with Tukey honest significant difference post hoc tests. RESULTS:Vancomycin did not cause significant changes in tenocyte viability after 2 and 6 hours of incubation at any concentration between 0 and 12,800 µg/mL. Incubation with vancomycin for 24 hours led to a significant decrease in cell viability at higher concentrations. CONCLUSION: Tenocytes derived from human patellar tendons exposed to relatively high concentrations of vancomycin for short periods of time do not demonstrate significant cell death and toxicity. CLINICAL RELEVANCE: Exposing tendons to vancomycin for a short period of time, such as before ACL reconstruction, is not likely to cause tenocyte toxicity because of vancomycin administration.
Authors: Alexander Themessl; Felix Mayr; Kate Hatter; Marco-Christopher Rupp; Jonas Pogorzelski; Andreas B Imhoff; Stefan Buchmann Journal: Knee Surg Sports Traumatol Arthrosc Date: 2021-12-06 Impact factor: 4.114