title: An investigation into the role of oxygen free radical scavengers in preventing polymethylmethacrylate-induced necrosis in an osteoblast cell culture.
abstract: This study examined the effect of polymethylmethacrylate (PMMA) on osteocytic necrosis and the role of free radical scavengers in minimizing this damage. Bovine osteoblast cells with a characteristic phenotype were seeded at a density of 4x10(4) cells/cm2 and cultured in a DMEM supplemented with 10% fetal calf serum. A transwell insert with 2 cc of PMMA was suspended above the culture, and a time log response curve was established following elusion of free radicals around the osteoblast media. Chemiluminescence was used to determine quantitative free radical release. Using a Student's two-tailed t test there was a significant difference in the amount of hydroxyl radical released at 1-6 hours compared with controls (P=.028). Using histologic markers, there was a significant correlation between the use of PMMA and osteoblast cell necrosis. Transwell plates were coated with varying concentrations of mannitol, a known hydroxyl radical scavenger. A log dose response curve was established. There was a clear statistical association between a 10% mannitol solution and a reduction in the free radical release from PMMA (P=.03). Similarly, using Trypan blue histologic staining, there was a significant reduction in PMMA-induced cell necrosis when 10% mannitol was used as a scavenger (P=.01). A Rockwell superficial hardness test was used to determine whether mannitol had any effect on the surface hardness of the polymer. No statistical difference could be found between those treated with mannitol and controls at a depth of up to 1 mm. These results demonstrate hydroxyl radical is released from the polymerization reaction of PMMA. These radicals cause cell death in an osteoblast culture medium. This has been addressed using a 10% mannitol solution, which reduced cell necrosis.
authors.affiliation: Department of Orthopedic Surgery, Mater Misericordiae Hospital, Dublin, Ireland.
authors.forename: J G
authors.initials: JG
authors.lastname: Kennedy
authors.forename: P
authors.initials: P
authors.lastname: O'Grady
authors.forename: D R
authors.initials: DR
authors.lastname: McCarthy
authors.forename: S J
authors.initials: SJ
authors.lastname: Johnson
authors.forename: D
authors.initials: D
authors.lastname: Hynes
authors.forename: M
authors.initials: M
authors.lastname: Walsh
authors.forename: F M
authors.initials: FM
authors.lastname: McManus
authors.forename: E
authors.initials: E
authors.lastname: Salih
authors.forename: M
authors.initials: M
authors.lastname: Gouverneur
authors.forename: J
authors.initials: J
authors.lastname: Fitzpatrick
mesh_terms: D002478:Cells, Cultured; D016166:Free Radical Scavengers / Q000494:pharmacology*; D008163:Luminescent Measurements; D008353:Mannitol / Q000494:pharmacology; D009336:Necrosis; D010006:Osteoblasts / Q000473:pathology*; D019904:Polymethyl Methacrylate / Q000494:pharmacology*; D017382:Reactive Oxygen Species
chemical_list: D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D008353:Mannitol; D019904:Polymethyl Methacrylate