Abstract
Purpose: Extruded tissue specimens excised during surgery on human intervertebral disc hernia and chondrocytes established and cultured from the excised tissue were observed via electron microscopy. Macrophages confirmed by CD68 immunostaining were added to the chondrocyte culture, and observed via electron microscopy. To observe, using an electron microscope, disc hernia degeneration at the cellular level as expressed in extruded tissue from a human intervertebral disc and cultured chondrocytes, and to investigate the mechanism of spontaneous regression and the effects of macrophages.
Materials and Methods: Tissues excised during surgery were fixed in various fixatives for electron microscopy and immune electron microscopy to avoid divided and treated with collagenase, and chondrocytes were then isolated and cultured. Human heparinized peripheral blood was separated using Ficoll and cultured. After culture, macrophages were collected and confirmed by CD68 immunostaining. These macrophages were added to the chondrocyte culture and observed under an electron microscope.
Results: Chondrocytes in the hernia is extruded region markedly differed from cultured chondrocytes. The tissue extruded from the intervertebral disc showed obvious degeneration such as changes in osmotic pressure. Macrophages were observed as the mechanism of spontaneous regression.
Discussion: In a previous study, we used ELISA to measure MMP-3 levels and TIMP-1 levels in both mRNA and serum in patients, and found a correlation between the two. In this study, we observed the pathological state of a disc hernia at the cellular level. When chondrocytes from the same tissue were cultured under conditions similar to those in the intervertebral disc, the extruded tissue showed a clear difference. It was considered that membrane osmotic pressure affects intervertebral disc hernia in humans and that protein transmission occurs in endoplasmic reticulum. It was also considered that spontaneous regression is due to the infiltration of macrophages.
The abstracts were prepared by Professor Jegan Krishnan. Correspondence should be addressed to him at the Flinders Medical Centre, Bedford Park 5047, Australia.
