Spatiotemporal Expression of 3-B-3(-) and 7-D-4 Chondroitin Sulfation, Tissue Remodeling, and Attempted Repair in an Ovine Model of Intervertebral Disc Degeneration

Cartilage. 2020 Apr;11(2):234-250. doi: 10.1177/1947603519876354. Epub 2019 Oct 3.

Abstract

Objective: Examination of intervertebral disc (IVD) regeneration in an ovine annular lesion model.

Hypothesis: Sulfation motifs are important functional determinants in glycosaminoglycans (GAGs). Previous studies have correlated 3-B-3(-) and 7-D-4 chondroitin sulfate (CS) motifs in tissues undergoing morphogenetic transition in development. We hypothesize that these motifs may also be expressed in degenerate IVDs and may represent a reparative response.

Design: Induction of disc degeneration by 5 mm or 6 × 20 mm lesions in the annulus fibrosus (AF) over 6 or 3 to 6 months postoperation (PO). Tissue sections were stained with toluidine blue-fast green, 3-B-3(-) and 7-D-4 CS-sulfation motifs were immunolocalized in 3-month PO 6 × 20 mm lesion IVDs. Sulfated glycosaminoglycan (GAG), 3-B-3(-), and 7-D-4 epitopes were quantitated by ELISIA (enzyme-linked immunosorbent inhibition assay) in extracts of AF (lesion site and contralateral half) and nucleus pulposus (NP) 0, 3, and 6 months PO.

Results: Collagenous overgrowth of lesions occurred in the outer AF. Chondroid metaplasia in ~20% of the 6 × 20 mm affected discs resulted in integration of an outgrowth of NP tissue with the inner AF lamellae preventing propagation of the lesion. 3-B-3(-) and 7-D-4 CS sulfation motifs were immunolocalized in this chondroid tissue. ELISIA quantified CS sulfation motifs demonstrating an increase 3 to 6 months PO in the AF lesion and a reduction in sulfated GAG not evident in the contralateral AF.

Conclusions: (1) Outer annular lesions underwent spontaneous repair. (2) Chondroid metaplasia of the inner 6 × 20 mm defect prevented its propagation suggesting an apparent reparative response.

Keywords: 3-B-3(−); 7-D-4; annular repair; chondroitin sulfate motifs; tissue regeneration; tissue remodeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Annulus Fibrosus / physiopathology
  • Chondroitin Sulfates / metabolism*
  • Disease Models, Animal
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes
  • Glycosaminoglycans / metabolism*
  • Intervertebral Disc / physiopathology*
  • Intervertebral Disc Degeneration / physiopathology*
  • Regeneration / physiology*
  • Sheep

Substances

  • Epitopes
  • Glycosaminoglycans
  • A73025
  • Chondroitin Sulfates