Abstract
Introduction
Discogenic pain is associated with ingrowth of blood vessels and nerves, but uncertainty over the extent of ingrowth is hindering development of appropriate treatments. We hypothesise that adult human annulus fibrosus is such a dense crosslinked tissue that ingrowth via the annulus is confined to a) peripheral regions, and b) fissures extending into the annulus.
Methods
Disc tissue was examined from 61 patients (aged 37–75 yrs) undergoing surgery for disc herniation, degeneration or scoliosis. 5 µm sections were stained with H&E to identify structures and tissue types. 30 µm frozen sections were examined using confocal microscopy, following immunostaining for CD31 (an endothelial cell marker), PGP 9.5 and Substance P (general and nociceptive nerve markers, respectively). Fluorescent tags were attached to the antibodies. ‘Volocity’ software was used to calculate numbers and total cross-sectional area of labelled structures, and to measure their distance from the nearest free surface (disc periphery, or annulus fissure).
Results
Maximum penetration of blood vessels and nerves from the peripheral annulus was 4,800 µm and 2,200 µm respectively. Maximum distance of nerves and vessels from the nearest free surface was 236 µm and 888 µm. Substance P (but not PGP 9.5) was co-localised with blood vessels, and both number and area of Substance P-stained structures were inversely correlated with grade of disc degeneration.
Interpretation
Thick sections and fluorescent markers can show reliably where labelled structures are not present. Results therefore support our hypothesis: deep penetration of nerves into the human annulus occurs only if fissures are present.
No conflicts of interest
No funding obtained
This abstract has not been previously published in whole or in part; nor has it been presented previously at a national meeting.
