Objective: To investigate the histopathological and immunohistochemical properties of degenerative changes in the ligamentum flavum of the cervical spine with calcium crystal deposition.
Methods: Sections of the calcified ligamentum flavum harvested from 26 patients who required cervical decompression were examined by scanning electron microscopy (SEM), energy dispersive X-ray microanalysis, immunohistochemical staining [for transforming growth factor (TGF)-Beta, vascular endothelial growth factor (VEGF), Sox9, and Msx2] and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labelling (TUNEL) method (for cell apoptosis).
Results: Energy dispersive x-ray microanalysis and SEM confirmed the deposited calcium to be calcium pyrophosphate dihydrate (CPPD) crystals. The calcified ligamentum flavum showed disorganisation of the elastic fibre bundles together with increased collagen fibrils in the matrix. Abundant hypertrophic chondrocytes were noted around the calcified lesions, which were strongly immunoreactive to TGF-Beta and VEGF. Staining for Sox9 was positive in metaplastic chondrocytes but negative in hypertrophic chondrocytes. Both chondrocytes and mesenchymal cells were positive for Msx2. TUNEL-positive hypertrophic chondrocytes were significantly more noticeable in nodular than diffusely scattered type of CPPD deposition.
Conclusions: Calcium crystal deposition in the cervical ligamentum flavum seems to progress with reduction in elastic fibres, increase in collagen fibrils in the matrix, and migration of metaplastic hypertrophic chondrocytes, whose differentiation is controlled by cytokines and transcriptional factors, and potentially regulate crystal formation. The presence of abundant TUNEL-positive hypertrophic chondrocytes around CPPD deposition suggests that materials from apoptotic cells play some role in crystal deposition.