Eric Röhner1, Timo Zippelius2, Sabrina Böhle2, Sebastion Rohe2, Georg Matziolis2, Benjamin Jacob2. 1. Department of Orthopedics, Jena University Hospital, Campus Eisenberg, Klosterlausnitzer Straße 81, 07607, Eisenberg, Germany. e.roehner@waldkliniken-eisenberg.de. 2. Department of Orthopedics, Jena University Hospital, Campus Eisenberg, Klosterlausnitzer Straße 81, 07607, Eisenberg, Germany.
Abstract
INTRODUCTION: Vancomycin powder (VP) is a well-established topical antibiotic used in spinal surgery to prevent surgical site infections. More recently its extension to hip and knee arthroplasty was introduced. The aim of this study was to examine toxic effects of VP on the viability of human chondrocytes. Our hypothesis was that VP damages human chondrocytes in vitro with increasing concentration and length of exposure. MATERIAL AND METHODS: Primary human chondrocytes were isolated and cultured from donated human knee joints. VP was added to these cultures with increasing concentrations (0-50 mg/ml) and length of exposure (0-336 h). Toxicity and viability were analyzed using LDH und XTT Elisa assays. Cell structure and determination of vital versus dead cells were visualized using light microscopy and fluorescence microscopy. RESULTS: Light microscopy and fluorescence microscopy visualized defect cell structures and cell death proportional to increasing dose and length of exposure to VP. The analysis of LDH activity data showed toxic effects on chondrocytes as early as 2,5 min after exposure to VP. XTT activity data revealed a significant toxic threshold of a VP concentration above 12.5 mg/ml. CONCLUSIONS: These results show that exposure to high VP concentrations yields to a damage of human chondrocytes in vitro. Chondrotoxicity is an immediate effect that is proportional to VP concentration. Therefore, the intraarticular use of high concentrations of vancomycin powder in the presence of native cartilage tissue must be considered critically.
INTRODUCTION:Vancomycin powder (VP) is a well-established topical antibiotic used in spinal surgery to prevent surgical site infections. More recently its extension to hip and knee arthroplasty was introduced. The aim of this study was to examine toxic effects of VP on the viability of human chondrocytes. Our hypothesis was that VP damages human chondrocytes in vitro with increasing concentration and length of exposure. MATERIAL AND METHODS: Primary human chondrocytes were isolated and cultured from donated human knee joints. VP was added to these cultures with increasing concentrations (0-50 mg/ml) and length of exposure (0-336 h). Toxicity and viability were analyzed using LDH und XTT Elisa assays. Cell structure and determination of vital versus dead cells were visualized using light microscopy and fluorescence microscopy. RESULTS: Light microscopy and fluorescence microscopy visualized defect cell structures and cell death proportional to increasing dose and length of exposure to VP. The analysis of LDH activity data showed toxic effects on chondrocytes as early as 2,5 min after exposure to VP. XTT activity data revealed a significant toxic threshold of a VP concentration above 12.5 mg/ml. CONCLUSIONS: These results show that exposure to high VP concentrations yields to a damage of human chondrocytes in vitro. Chondrotoxicity is an immediate effect that is proportional to VP concentration. Therefore, the intraarticular use of high concentrations of vancomycin powder in the presence of native cartilage tissue must be considered critically.
Entities:
Keywords:
Cell death; Human chondrocytes; Toxicity; Vancomycin powder
Authors: Matthew W Bullock; Matthew L Brown; Daniel N Bracey; Maxwell K Langfitt; John S Shields; Jason E Lang Journal: J Arthroplasty Date: 2016-11-22 Impact factor: 4.757