Matrix remodeling during intervertebral disc growth and degeneration detected by multichromatic FAST staining.
Various imaging techniques have been used to assess degeneration of the intervertebral disc, including many histological methods, but cartilage-oriented histological stains do not clearly show the comparatively complex structures of the disc. In addition, there is no integrated method to assess efficiently both the compartmental organization and matrix composition in disc samples. In this study, a novel histological method, termed FAST staining, has been developed to investigate disc growth and degeneration by sequential staining with fast green, Alcian blue, Safranin-O, and tartrazine to generate multichromatic histological profiles (FAST profiles). This identifies the major compartments of the vertebra-disc region, including the cartilaginous endplate and multiple zones of the annulus fibrosus, by specific FAST profile patterns. A disc degeneration model in rabbit established using a previously described puncture method showed gradual but profound alteration of the FAST profile during disc degeneration, supporting continual alteration of glycosaminoglycan. Changes of the FAST profile pattern in the nucleus pulposus and annulus fibrosus of the postnatal mouse spine suggested matrix remodeling activity during the growth of intervertebral discs. In summary, we developed an effective staining method capable of defining intervertebral disc compartments in detail and showing matrix remodeling events within the disc. The FAST staining method may be used to develop a histopathological grading system to evaluate disc degeneration or malformation.
Leung
Victor Y L
VY
Department of Biochemistry, The University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong, China.
Chan
Wilson C W
WC
Hung
Siu-Chun
SC
Cheung
Kenneth M C
KM
Chan
Danny
D
D000423:Alcian Blue; D000818:Animals; D000831:Animals, Newborn; D005109:Extracellular Matrix / Q000473:pathology*; D005260:Female; D007403:Intervertebral Disc / Q000254:growth & development* / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010619:Phenazines; D011817:Rabbits; D012394:Rosaniline Dyes; D013122:Spinal Diseases / Q000473:pathology*; D013194:Staining and Labeling; D013645:Tartrazine
D010619:Phenazines; D012394:Rosaniline Dyes; C035906:Fast Green; D013645:Tartrazine; D000423:Alcian Blue; C009195:safranine T