J Korean Neurosurg Soc.  2022 Mar;65(2):186-195. 10.3340/jkns.2021.0082.

Histological Changes of Cervical Disc Tissue in Patients with Degenerative Ossification

• Xiong Y
• Yang YL
• Gao YS
• Wang XM
• Yu X

• KMID: 2527175
• DOI: http://doi.org/10.3340/jkns.2021.0082

Abstract

Objective
: To explore the histological feature of the cervical disc degeneration in patients with degenerative ossification (DO) and its potential mechanisms.
Methods
: A total of 96 surgical segments, from cervical disc degenerative disease patients with surgical treatment, were divided into ossification group (group O, n=46) and non-ossification group (group NO, n=50) based on preoperative radiological exams. Samples of disc tissues and osteophytes were harvested during the decompression operation. The hematoxylin-eosin staining, Masson trichrome staining and Safranin O-fast green staining were used to compare the histological differences between the two groups. And the distribution and content of transforming growth factor (TGF)-β1, p-Smad2 and p-Smad3 between the two groups were compared by a semi-quantitative immunohistochemistry (IHC) method.
Results
: For all the disc tissues, the content of disc cells and collagen fibers decreased gradually from the outer annulus fibrosus (OAF) to the central nucleus pulposus (NP). Compared with group NO, the number of disc cells in group O increased significantly. But for proteoglycan in the inner annulus fibrosus (IAF) and NP, the content in group O decreased significantly. IHC analysis showed that TGF-β1, p-Smad2, and p-Smad3 were detected in all tissues. For group O, the content of TGF-β1 in the OAF and NP was significantly higher than that in group NO. For p-Smad2 in IAF and p-Smad3 in OAF, the content in group O were significantly higher than group NO.
Conclusion
: Histologically, cervical disc degeneration in patients with DO is more severe than that without DO. Local higher content of TGF-β1, p-Smad2, and p-Smad3 are involved in the disc degeneration with DO. Further studies with multi-approach analyses are needed to better understand the role of TGF-β/Smads signaling pathway in the disc degeneration with DO.

Keyword

Intervertebral disc degeneration; Ossification, heterotopic; Histological techniques; Transforming growth factor beta; Smad proteins

Figure

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  Fig. 1. The DO around the surgical segments in group O. A and B : Preoperative X-ray and computed tomographic (CT) examination showed that the osteophytes were found in front of the C5-6 and C6-7, respectively. C and D : Preoperative X-ray and CT examination showed the osteophytes were found both in front and behind C5-6.

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  Fig. 2. General histological staining of the osteophyte. A : Hematoxylin-eosin (HE) stain, ×100. B : HE stain, ×400, the black arrow indicates osteoblasts, and the white arrow indicates osteoclasts. C : Modified Masson trichrome stain, ×100. D : Modified Safranin O-fast green cartilage stain, ×100.

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  Fig. 3. General histological staining and semi-quantitative comparison of the disc tissue. A : Hematoxylin-eosin (HE) staining, ×100. B : Comparison of cell counts in disc tissues. C : Modified Masson trichrome staining, ×100. D : Comparison of collagen fiber content. E : Modified Safranin O-fast green cartilage staining, ×200. F : Comparison of proteoglycan content. *p<0.05. †p<0.001. Group O : ossification group, group NO : non-ossification group, OAF : outer annulus fibrosus, IAF : inner annulus fibrosus, NP : nucleus pulposus, AOD : average optical density

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  Fig. 4. Immunohistochemistry (IHC) staining and semi-quantitative comparison of the disc tissue. A, C, and E : The IHC staining of TGF-β1, p-Smad2, and p-Smad3, ×200. B, D, and F : Comparison of the AOD of TGF-β1, p-Smad2, and p-Smad3, respectively. *p<0.05. †p<0.001. TGF : transforming growth factor, group O : ossification group, group NO : non-ossification group, OAF : outer annulus fibrosus, IAF : inner annulus fibrosus, NP : nucleus pulposus, AOD : average optical density.

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  Fig. 5. Immunohistochemistry staining of the osteophyte, ×200. Black arrows showed osteoblasts and white arrows showed osteocytes. A : For transforming growth factor-β1. B : For p-Smad2. C : For p-Smad3.

Reference

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