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NF-B Activation and iNOS Expression in the Synovial Membrane of Rat Temporomandibular Joints after Induced Synovitis

¹ Departments of Oral Anatomy and Cell Biology,
² Removable Prosthodontics, and
³ Orthodontics, Kyushu University Graduate School of Dental Science, Fukuoka 812-8582, Japan;

View larger version (100K): [in this window] [in a new window]   Figure 1. A schematic illustration and light micrographs of the rat TMJ. (a) A schematic diagram of sagittal sections of the central portion of the rat TMJ. The rat TMJ is divided into upper and lower compartments (UC, LC) by an articular disc, and the synovial membrane lines these compartments. The boxes indicate the observed portions of each section. Boxed numbers correspond to the respective Fig. numbers, and boxed letters (A to D) correspond to the portions observed in the histopathological analysis. Light micrographs of 10-µm sagittal sections of the (b,d) anterior and (c,e) posterior portions in induced-synovitis and control rat TMJs. In the induced-synovitis TMJ (b,c), the synovium consisting of 7 cell layers or more (grade 2, G2) is seen in the anterior (b) and posterior (c) portions. Vascular dilation, fibrin deposition (asterisk), and fibrous thickening are also observed. C, mandibular condyle; V, blood vessel. In the control TMJ (d,e), 1-3 layers (grade 0, G0) face the upper and lower joint compartments. No inflammatory change is found in the control synovium. Hematoxylin and eosin staining (b-e). Original magnification: x400 (b-e). Bar = 25 µm (b-e).

View larger version (117K): [in this window] [in a new window]   Figure 2. Light micrographs of serial sagittal sections of the anterior portion in induced-synovitis (a,b) and control (c,d) rat TMJs immunostained with either anti-NF-B (a,c) or anti-iNOS (b,d) antibody. NF-B immunoreactivity is increasingly expressed in the cytoplasm of the synovial lining cells of the 7 or more layers (grade 2, G2) in the induced-synovitis TMJ (a) compared with that of the 1 or 3 layers (grade 0, G0) in the control TMJ (c), but is not shown in the nuclei of the synovial cells in the experimental or control TMJs. C, mandibular condyle; LC, lower joint compartment; V, vein. The immunoreaction for iNOS is also increasingly seen in the synovial lining cells of the 1-3 layers in the experimental TMJ (b) compared with the control TMJ (d). Original magnification: x100 (a-d). Bar = 100 µm (a-d).

View larger version (108K): [in this window] [in a new window]   Figure 3. Light micrographs of sagittal sections of the posterior region in induced-synovitis (a,c-e) and control (b) rat TMJs with the use of Southwestern histochemistry for NF-B. In the synovitis TMJ, strong reaction for NF-B (arrowhead) is seen in the 4-6 synovial cell layers (grade 1, G1). Inset: Higher magnification of the area indicated by the arrow in Fig. 3a. NF-B is strongly expressed in the nuclei (N) of the synovial cells. F, synovial fold; UC, upper joint compartment; V, vein. In the control TMJ, NF-B is weakly expressed in the nuclei of the synovial lining cells of the 1-3 cell layers (grade 0, G0) (arrowhead). Inset: Higher magnification of the area indicated by the arrow in Fig. 3b. (c) Strong staining for the DIG-labeled ds-ODN probe is found in the surface cells of the synovial membrane (arrowheads) (probe +). JC, joint compartment. (d) When non-labeled ds-ODN (50-fold) was incubated competitively with ds-ODN probe (Comp.), NF-B reactivity in the synovium decreased or disappeared (arrowhead). (e) No reaction is observed in the synovial membrane stained with the pre-incubation buffer without ds-ODN probe (probe -) (arrowhead). Eosin staining. Original magnification: x400, Inset: x3600 (a,b), x240 (c-e). Bar = 25 µm (a,b), 15 µm (c-e).

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