BIOCOMPATIBILITY OF ALLOGRAFT FOR IMPACTION GRAFTING: EFFECT OF MARROW REMOVAL.

Abstract

Impaction grafting with morsellised allograft is becoming the treatment of choice for revision arthroplasty, especially in the younger patient. The optimum treatment of the graft prior to impaction has not been determined. Some groups wash the graft prior to impaction and others do not. Washing of graft has been shown to enhance bone ingrowth in an 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.

Paired samples of washed and unwashed morcellised FFH allograft were taken during revision hip arthroplasties. Washing was performed by 4 consecutive rinses in 300ml warmed saline, the bone being filtered between each exchange of saline. Contact cytotoxicity assays involved culture of cell lines in direct contact with bone samples. Quantitative cytotoxicity assays utilised culture media conditioned with the bone samples and subsequent assessment of cell metabolism and viability using both dimethylthiazol (MTT) and neutral red (NR) assays. Assays were performed with human osteoblastic (MG63) and fibroblastic (HSF) cell lines.

Nine pairs of samples were analysed. Contact assays demonstrated a clear zone of cellular inhibition around the unwashed bone samples. Quantitative 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 significance (p=0.06).

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.