title: An evaluation of the capacity of differently prepared demineralised bone matrices (DBM) and toxic residuals of ethylene oxide (EtOx) to provoke an inflammatory response in vitro.
abstract: Demineralised bone matrix (DBM) is a form of allogeneic tissue graft widely used in oral and maxillofacial procedures. There is a long history of controversy relating to the suitability of ethylene oxide gas (EtOx) as a terminal sterilisation agent for this graft, relating to its effects on the clinical performance of the grafts. Furthermore, the generation of a toxic residual chemical (ethylene chlorohydrin, ECl) during the ethylene oxide sterilisation of patellar tendon allografts has been implicated in the failure of these grafts owing to the induction of a localised inflammatory response. In this study we have investigated the capacity of a range of different DBM preparations, and ECl dilutions, to induce the production of three pro-inflammatory cytokines, interleukin-6 (IL-6), interleukin-1beta (IL-1beta), and tumour necrosis factor alpha (TNF-alpha) from human peripheral blood mononuclear cells (PBMNCs). The levels of EtOx and ECl in EtOx terminally sterilised DBM and mineralised bone grafts were measured by gas chromatography. It was found that the only factor capable of rendering DBM pro-inflammatory was the presence of small (<20 micrometre diameter) DBM particles. No other processing or sterilisation technique resulted in the DBM becoming pro-inflammatory. Although it was also found that DBM, when EtOx-sterilised, retained more ECI than mineralised bone grafts following a standard EtOx sterilisation protocol, ECl did not provoke an inflammatory response in vitro at levels up to and including those which are cytotoxic to PBMNCs.
authors: Yorkshire Regional Tissue Bank, National Blood Service, Pinderfields General Hospital, Wakefield, UK. rjlomas@aol.com
authors: R J
authors: RJ
authors: Lomas
authors: H L
authors: HL
authors: Gillan
authors: J B
authors: JB
authors: Matthews
authors: E
authors: E
authors: Ingham
authors: J N
authors: JN
authors: Kearney
mesh_terms: D018775:Bone Demineralization Technique; D001842:Bone and Bones / Q000378:metabolism*; D002849:Chromatography, Gas; D018920:Coculture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005027:Ethylene Oxide / Q000633:toxicity*; D006801:Humans; D007249:Inflammation / Q000139:chemically induced*; D018836:Inflammation Mediators / Q000378:metabolism; D007375:Interleukin-1 / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis; D009000:Monocytes / Q000378:metabolism; D010316:Particle Size; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D018836:Inflammation Mediators; D007375:Interleukin-1; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D005027:Ethylene Oxide