A scanning electron microscopic study of in vitro toxicity of ethylene-oxide-sterilized bone repair materials.

Polylactic acid (PLA) and polyglycolic acid (PGA) have been under investigation for use in the management of hard- and soft-tissue wounds. Current research has included the incorporation of osteo-inductive substances into a PLA-PGA copolymer alloplastic implant material for enhancement of the healing of osseous defects. Conventional methods of sterilization--such as dry heat, steam heat, or 60Co--tend either to destroy or attenuate osteo-inductive activity and alter polymer biodegradation. Ethylene oxide (EO) gas sterilization is currently being tested as an alternate method. This study examined the relationship of EO-induced cytotoxicity to the length of time of polymer aeration following EO sterilization. Three groups of copolymer implant discs were studied: (1) 50:50 PLA-PGA copolymer, (2) PLA-PGA polymer with hydroxyapatite (HA), and (3) PLA-PGA with autolyzed, antigen-extracted (AA) bone particles. Polymer discs, as well as particulate HA and AA bone controls, were sterilized with EO for 12 hours. Following periods of two weeks, one week, one day, or no subsequent vacuum aeration, samples were placed into 24-well culture plates. A suspension of human fibroblasts was added to each well. Cell growth and attachment were permitted for 24 hours. Medium was then removed, and solutions for cell fixation, buffer washing, and dehydration were added to each well. SEM examination revealed changes in cell growth with increasing periods of aeration suggestive of increasing cell vitality. Cells growing on discs having no aeration were small, round, and lobulated, whereas those of seven to 14 days' aeration were more numerous, and flattened with many microvilli, pseudopodia, and dendritic processes, features consistent with normal cell morphology. These results suggest that EO-sterilized polymer implants should be aerated for least seven to 14 days prior to surgical use.