Local antibiotics released through a carrier is a commonly used technique to prevent infection in orthopaedic procedures. An interesting carrier in aseptic bone reconstructive surgery are bone chips impregnated with AB solution. Systemically administered Cefazolin (CFZ) is used for surgical site infection prophylaxis however in vitro study showed that fresh frozen and processed bone chips impregnated with CFZ solution completely release the CFZ within a few hours. On the other hand irradiated freeze-dried bone chips, treated with supercritical CO2 (scCO2) have been shown to be an efficient carrier for the antibiotics vancomycine or tobramycine. With this pilot study we wanted to investigate if CFZ solution impregnation of bone chips treated with scCO2 shows a more favorable release pattern of CFZ. The bone chips were prepared using the standard scCO2 protocol and were impregnated with 100 mg/ml cefazolin at different timepoints during the process: before freeze drying (BC type A), after freeze drying (BC type B) and after gamma-irradiation. 0.5g of the impregnated bone grafts were incubated with 5ml of fetal calf serum (FCS) at 37°C. At 2, 4, 6, 8 and 24h of incubation 200µl of eluate was taken for analysis. After 24h the remaining FCS was removed, bone grafts were washed and new FCS (5ml) was added. Consecutive eluate samples were taken at 48, 72 and 96h of incubation. The concentration of CFZ in the eluates was measured with the validated UPLC-DAD method. Analysis was performed in triplicate.Aim
Method
Vancomycin -impregnated bonechips from a human morselized femoral head allograft (BCs) are used in orthopaedic surgery to treat infections. Literature suggests that bonechips can be efficient vancomycin carriers, but due to the diversity in the type of bonechips, of impregnation and of method used to evaluate AB release, there are no uniform guidelines. We performed an in vitro study to examine the release of vancomycin from solution-impregnated deepfrozen processed bonechips. Quantification was performed using a fully validated chromatographic method. Results were compared with the elution-profile from Osteomycin®, a commercially available lyophilised processed bonegraft. Different vancomycine impregnation-concentrations and impregnation-durations of frozen processed bonechips were investigated. After impregnation, bonechips were rinsed with saline in order to determine only the absorbed vancomycin. Elution was performed in newborn calf serum at 37°C. Eluted vancomycin concentrations were determined using Ultra Performance Liquid Chromatography – Diode Array Detection (UPLC-DAD). In addition an elution study was performed on the commercially available Osteomycin®, bone chips containing vancomycine. Using processed frozen bonechips, an impregnation-concentration of minimum 100 mg/mL during 10 minutes delivers the desired local concentration (therapeutic window 25 – 1000 mg/L) for 3 days. Longer impregnation time at this concentration had no effect. Osteomycin®: delivers the desired local concentration for 8 days in our experimental setting. Literature suggests that freshfrozen BCs can be used as carrier for vancomycin through solution-impregnation [1,3]. There is however much less information on the carrier-capacities of frozen processed bonechips, a type used in our hospital. Our impregnation-protocol was based on that of Mathyssen et al., but with direct quantification of elution concentrations. Impregnation with vancomycine 100 mg/mL during 10 minutes results in a release above the desired concentration for 3 days, which seems too short when treating bone-infections. Osteomycin®, shows a substantially longer elution [2]. Vancomycin-solution impregnation of frozen processed bonechips may not be sufficient to obtain the desired release-characteristics for the treatment of bone-infections.
