Abstract
Fresh frozen femoral head (FFH) allograft is commonly used in impaction grafting for revision hip arthroplasty and long term success has been demonstrated by some groups. The optimum treatment of the graft prior to impaction has not yet been determined. Some groups wash the graft prior to impaction and others do not. Washing of the graft has been shown to improve bone ingrowth in a bone chamber animal model however the reasons for this remain unclear. The aim of this study was to identify any underlying cellular cytotoxicty of fresh frozen allograft bone before and after washing.
Samples of morcellised FFH allograft were taken during revision hip arthroplasties prior to impaction grafting. Paired samples, taken before and after washing were taken from each case. Washing was performed by 4 consecutive washes in 300ml warmed saline, the bone being filtered between each exchange of saline. Cytotox-icity was assessed for all samples using both contact and extract assays. Contact assays involved culture of cell lines in direct contact with bone samples. Extract assays utilised culture media conditioned with bone samples and subsequent quantitative assessment of cell metabolism and viability using both dimethylthiazol (MTT) and neutral red (NR) assays. All assays were performed using both human osteoblastic (MG63) and fibroblastic (HSF) cell lines.
Nine pairs of samples were analysed for cytotoxicity using both cell lines. Contact assays demonstrated a clear zone of cellular inhibition around the unwashed bone samples. Extract assays were performed in triplicate for each cell type and both MTT and NR assays giving 108 paired assay results. 88.9% of pairs (92/108) showed cytotoxicity in the unwashed sample. No washed samples demonstrated cytotoxicity. When grouped by assay and cell type, analysis of means showed statistically significant differences between washed and unwashed samples in MG63-NR (p=0.0025), HSF-NR (p=0.0004) and MG63-MTT (p=0.008). The difference observed in the HSF-MTT assays did not reach statistical signifi-cance (p=0.06).
In conclusion, we have shown that unwashed FFH allograft can be cytotoxic to human osteoblastic and fibroblastic cell lines in vitro. This suggests that allograft should be washed prior to impaction in order to optimise the biological compatibility.
Correspondence should be addressed to Mr Carlos Wigderowitz, Senior Lecturer, University Department of Orthopaedic and Trauma Surgery, Ninewells Hospital and Medical School, Dundee DD1 9SY.
