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Dose Dependent Chondrotoxicity of Intrasite Vancomycin: a Porcine Model
Dillon Arango, MD1; Michael Narvaez, MD1; William Smith, MD1; James Raphael, MD1; Solomon Praveen Samuel, D. Eng1
1Einstein Medical Center, Philadelphia, PA, 2Temple University Hospital, Philadelphia, PA
Introduction: There is a recent increase in the use of intrasite vancomycin for the prevention of surgical site infections (SSI) not only in the spine yet also in trauma, tumor, and upper extremity surgery. No studies have explored the possible acute deleterious effects of intrasite application of vancomycin on cartilage. This ex-vivo study evaluated the effect of varying vancomycin concentrations on porcine cartilage explants.
Methods: After prepping the knee with isopropyl alcohol, 3 mm punch biopsy cartilage explants were harvested from the stifle joints of Yorkshire pig cadavers within 1 hour of being euthanized (Figure 1). Explants were washed with normal saline and kept in standard cell culture media (DMEM media supplemented with 10% fetal bovine serum) for 24 hours and then subjected to various concentrations of vancomycin for one hour. Groups included vancomycin at 50, 25, 10, 5, 1, 0.5, 0.05, .01, 0.005, 0.0 mg/ml. After drug exposure, samples were washed with saline and tissue metabolism was evaluated with resazurin (Figure 2) followed by addition of a tetrazolium compound (MTS). After 2 hours, the optical density of the solutions was measured in order to quantify cell metabolism and viability. Analysis of variance (ANOVA) and Tukey range test were used to detect statistical differences between groups.
Results: Resazurin assay data collected 3 hours after vancomycin application demonstrated that vancomycin concentrations above 25 mg/ml resulted in a statistically significant decrease in cell metabolism (p<.001). Lower concentrations of vancomycin failed to demonstrate any significant difference in regards to cell metabolism. MTS assays performed <24 hours from vancomycin application demonstrated that cartilage explants exposed to 0.005 mg/ml resulted in 84% decrease in cell viability (p<.05). Explants in the 50 mg/ml group demonstrated a 99% decrease in cell viability (p<.05) (Figure 3).
Conclusion: Application of intrasite vancomycin significantly decreases chondrocyte viability in this ex-vivo model. With the increase in vancomycin use for wounds, not only in the spine, future studies should focus on in-vivo analysis of cartilage viability after application of vancomycin. Figure 1: Stifle joint of pig
 Figure 2: Resazurin cartilage metabolism evaluation. Changes in color from blue to reddish hue
 Figure 3: % of cell viability after vancomycin application
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