Ossification of the posterior longitudinal ligament of the cervical spine: histopathological findings around the calcification and ossification front

2007 ◽  
Vol 7 (2) ◽  
pp. 174-183 ◽  
Author(s):  
Ryuichiro Sato ◽  
Kenzo Uchida ◽  
Shigeru Kobayashi ◽  
Takafumi Yayama ◽  
Yasuo Kokubo ◽  
...  

Object. The authors studied the histological and immunohistochemical features of ossified posterior longitudinal ligament (PLL) of the cervical spine, especially in the calcification and ossification front. Methods. Samples of en bloc ossified PLL plaque obtained in 31 patients were stained with H & E and immuno-histochemically prepared for collagens (types I and II), vascular endothelial growth factor (VEGF), transforming growth factor (TGF)–β, and bone morphogenetic protein (BMP)–2, and by terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling method for apoptosis. Results. Enchondral ossification was evident between the ligamentous enthesis and deep layer of the ligament, with irregularly disorganized arrangement of elastic fibers in association with advancement of the degenerative process. In the ossification front, many hypertrophic metaplastic chondrocytes were noted in the ossifying plaque immediately contiguous to the ligament fibers, together with a considerable degree of neovascularization. Both TGFβ and BMP-2 were highly expressed in metaplastic hypertrophic chondrocytes in the ossification front, and BMP-2 was also expressed in fibroblastic cells near the ossified PLL plaque. Expression of type I collagen was significant in the matrix of the ossified PLL lesion, whereas that of type II was marked in metaplastic chondrocytes in the ossification front. Apoptotic hypertrophic chondrocytes were observed mainly in the fibrocartilaginous area near the calcification front. Conclusions. The enchondral ossification process in the ossified PLL was closely associated with degenerative changes of elastic fibers and cartilaginous cartilage formation, together with the appearance of metaplastic hypertrophic cartilage cells and neovascularization. The authors also found that VEGF-positive metaplastic chondrocytes in the ossification front and different expression patterns of collagens probably play some role in the extension of the ossified PLL from the ossification front.

2007 ◽  
Vol 7 (2) ◽  
pp. 184-193 ◽  
Author(s):  
Takafumi Yayama ◽  
Kenzo Uchida ◽  
Shigeru Kobayashi ◽  
Yasuo Kokubo ◽  
Ryuichiro Sato ◽  
...  

Object. The object of this study was to histopathologically and immunohistochemically characterize ossification of the ligamentum flavum (OLF) in samples of the thoracic spine harvested en bloc during surgery and to enhance the understanding of the ossifying process, particularly calcification and ossification. Methods. Samples of OLF plaque were obtained en bloc from 43 patients who underwent posterior decompression. The histopathological findings were correlated with radiological subtypes using computed tomography. The expression of type I and type II collagens, vascular endothelial growth factor (VEGF), transforming growth factor (TGF)β, and bone morphogenetic protein (BMP)–2 was investigated. Results. Surgical decompression using the posterior floating and en bloc resection technique resulted in neurological improvement in 40 of 43 patients. Progression of the OLF lesion longitudinally and medially was associated with significant degeneration of elastic fibers, fiber bundle derangement, decrements in fiber diameter, and fragmentation. Calcification and ossification paralleled the degeneration of the elastic fibers, extended more medially, and fused in the central area. Expression of BMP-2, TGFβ, and VEGF was significant in chondrocytes in the calcified cartilage and fibrocartilage layers, especially around the calcified front. Conclusions. Histopathologically, the progress of calcification and ossification was closely associated with the degeneration of elastic fibers and with significant expression of BMP-2, TGFβ, and VEGF in the ossification front.


Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 679
Author(s):  
Benedict-Uy Fabia ◽  
Joshua Bingwa ◽  
Jiyeon Park ◽  
Nguyen-Mihn Hieu ◽  
Jung-Hoon Ahn

Pseudomonas fluorescens, a gram-negative bacterium, has been proven to be a capable protein manufacturing factory (PMF). Utilizing its ATP-binding cassette (ABC) transporter, a type I secretion system, P. fluorescens has successfully produced recombinant proteins. However, besides the target proteins, P. fluorescens also secretes unnecessary background proteins that complicate protein purification and other downstream processes. One of the background proteins produced in large amounts is FliC, a flagellin protein. In this study, the master regulator of flagella gene expression, fleQ, was deleted from P. fluorescens Δtp, a lipase and protease double-deletion mutant, via targeted gene knockout. FleQ directs flagella synthesis, so the new strain, P. fluorescens ΔfleQ, does not produce flagella-related proteins. This not only simplifies purification but also makes P. fluorescens ΔfleQ an eco-friendly expression host because it will not survive outside a controlled environment. Six recombinant growth factors, namely, insulin-like growth factors I and II, beta-nerve growth factor, fibroblast growth factor 1, transforming growth factor beta, and tumor necrosis factor beta, prepared using our supercharging method, were successfully secreted by P. fluorescens ΔfleQ. Our findings demonstrate the potential of P. fluorescens ΔfleQ, combined with our supercharging process, as a PMF.


2001 ◽  
Vol 12 (3) ◽  
pp. 675-684 ◽  
Author(s):  
Jules J.E. Doré ◽  
Diying Yao ◽  
Maryanne Edens ◽  
Nandor Garamszegi ◽  
Elizabeth L. Sholl ◽  
...  

Transforming growth factor-βs (TGF-β) are multifunctional proteins capable of either stimulating or inhibiting mitosis, depending on the cell type. These diverse cellular responses are caused by stimulating a single receptor complex composed of type I and type II receptors. Using a chimeric receptor model where the granulocyte/monocyte colony-stimulating factor receptor ligand binding domains are fused to the transmembrane and cytoplasmic signaling domains of the TGF-β type I and II receptors, we wished to describe the role(s) of specific amino acid residues in regulating ligand-mediated endocytosis and signaling in fibroblasts and epithelial cells. Specific point mutations were introduced at Y182, T200, and Y249 of the type I receptor and K277 and P525 of the type II receptor. Mutation of either Y182 or Y249, residues within two putative consensus tyrosine-based internalization motifs, had no effect on endocytosis or signaling. This is in contrast to mutation of T200 to valine, which resulted in ablation of signaling in both cell types, while only abolishing receptor down-regulation in fibroblasts. Moreover, in the absence of ligand, both fibroblasts and epithelial cells constitutively internalize and recycle the TGF-β receptor complex back to the plasma membrane. The data indicate fundamental differences between mesenchymal and epithelial cells in endocytic sorting and suggest that ligand binding diverts heteromeric receptors from the default recycling pool to a pathway mediating receptor down-regulation and signaling.


1994 ◽  
Vol 269 (31) ◽  
pp. 20172-20178 ◽  
Author(s):  
H. Yamashita ◽  
P. ten Dijke ◽  
P. Franzén ◽  
K. Miyazono ◽  
C.H. Heldin

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