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.

Scanning electron microscopic study of cell attachment to biodegradable polymer implants.

The biodegradable polymers--polylactic acid (PLA) and polyglycolic acid (PGA)--are currently being studied as carriers for bioactive bone regeneration compounds. The inclusion of osteo-inductive substances in poly-(DL, lactide-co-glycolide) copolymer alloplastic implants has been shown to enhance the repair of osseous defects. The purpose of this study was to examine, by SEM, the attachment relationship of biodegradable polymer implants to cells and tissue matrix. Three groups of copolymer implants were studied: (1) plain 50:50 PLA-PGA copolymer, (2) PLA-PGA copolymer with hydroxyapatite (HA), and (3) PLA-PGA copolymer with autolyzed, antigen-extracted (AA) bone particles. Polymer discs were surgically implanted into the pectoralis muscles of rats. At seven, 14, and 21 days post-implantation, the baskets were removed and the contents prepared for SEM. Results showed that at one week, implants were coated primarily with red and white blood cells in a fibrinoid clot. Degradation of the polymers was evidenced by irregular enlarging of polymer surface pores. At two and three weeks, polymers became lobular and then fibrinoid as degradation progressed. Inflammatory cell and red blood cell adhesions were increasingly replaced by fibroblasts and collagen matrix deposition. As polymer degradation progressed, AA and HA particles were exposed; however, the lack of cell or tissue adhesion in these areas suggests that degradation may be more influenced by the fluid environment than by direct cell attachment. Furthermore, degradation may inhibit direct cell attachment.