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  • Annulus fissures are mechanically and chemically conducive to the ingrowth of nerves and blood vessels.

Annulus fissures are mechanically and chemically conducive to the ingrowth of nerves and blood vessels.

Spine (2012-06-19)
Manos Stefanakis, Maan Al-Abbasi, Ian Harding, Phillip Pollintine, Patricia Dolan, John Tarlton, Michael A Adams
ABSTRACT

Mechanical and biochemical analyses of cadaveric and surgically removed discs. To test the hypothesis that fissures in the annulus of degenerated human discs are mechanically and chemically conducive to the ingrowth of nerves and blood vessels. Discogenic back pain is closely associated with fissures in the annulus fibrosus, and with the ingrowth of nerves and blood vessels. Three complementary studies were performed. First, 15 cadaveric discs that contained a major annulus fissure were subjected to 1 kN compression, while a miniature pressure transducer was pulled through the disc to obtain distributions of matrix compressive stress perpendicular to the fissure axis. Second, Safranin O staining was used to evaluate focal loss of proteoglycans from within annulus fissures in 25 surgically removed disc samples. Third, in 21 cadaveric discs, proteoglycans (sulfated glycosaminoglycans [sGAGs]) and water concentration were measured biochemically in disrupted regions of annulus containing 1 or more fissures, and in adjacent intact regions. Reductions in compressive stress within annulus fissures averaged 36% to 46%, and could have been greater at the fissure axis. Stress reductions were greater in degenerated discs, and were inversely related to nucleus pressure (R(2) = 47%; P = 0.005). Safranin O stain intensity indicated that proteoglycan concentration was typically reduced by 40% at a distance of 600 μm from the fissure axis, and the width of the proteoglycan-depleted zone increased with age (P < 0.006; R(2) = 0.29) and with general proteoglycan loss (P < 0.001; R(2) = 0.32). Disrupted regions of annulus contained 36% to 54% less proteoglycans than adjacent intact regions from the same discs, although water content was reduced only slightly. Annulus fissures provide a low-pressure microenvironment that allows focal proteoglycan loss, leaving a matrix that is conducive to nerve and blood vessel ingrowth.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Gram′s safranin solution, for microscopy
Sigma-Aldrich
Gram-Hucker′s stain solution (III)
Sigma-Aldrich
Safranin O, certified by the Biological Stain Commission
Sigma-Aldrich
Safranin O, for microscopy (Bact., Bot., Hist.), indicator (pH 0.3-1.0)
Sigma-Aldrich
Safranin O, Dye content ≥85 %