J. -S. Kim, M. H. Ali, F. Wydra, X. Li, J. L. Hamilton, H. S. An, G

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Presentation transcript:

Characterization of degenerative human facet joints and facet joint capsular tissues J.-S. Kim, M.H. Ali, F. Wydra, X. Li, J.L. Hamilton, H.S. An, G. Cs-Szabo, S. Andrews, M. Moric, G. Xiao, J.H.-C. Wang, Di Chen, J.M. Cavanaugh, H.-J. Im Osteoarthritis and Cartilage Volume 23, Issue 12, Pages 2242-2251 (December 2015) DOI: 10.1016/j.joca.2015.06.009 Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Gross and histologic appearances of normal and degenerative FJs. A, Anatomical structure of human lumbar FJ and preparation of inferior articular process in lumbar FJ. B, Gross appearances of FJs and FJC tissues. Each FJ was assigned a representative grade for the appearance of the inferior facet. The higher graded FJ samples demonstrates an increase in the size of edge osteophytes, reactive bone, and cartilage eburnation. C, Safranin O, D, Alcian Blue Hematoxylin/Orange G, and E, Hematoxylin & Eosin staining of FJ cartilage show severe depletion of proteoglycan, fibrillation, structural and morphological changes in degenerative FJs (G4) compared to normal FJs (G0). The results represent donor number of at least n = 3 for G0 and n = 6 for each grades, G2-4. Osteoarthritis and Cartilage 2015 23, 2242-2251DOI: (10.1016/j.joca.2015.06.009) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Assessment of inflammation, neovascularization, and neuron ingrowth in normal and degenerative FJC tissue. A, Immunohistochemical (IHC) staining of CD11b indicates increased infiltration of inflammatory cells in degenerative FJC tissues (G3/4) compared to normal FJC tissues (G0). B, IHC staining of smooth muscle actin (a-SMA) shows markedly increased angiogenic events in degenerative FJC (G3/4) tissues compared to normal FJC tissues (G0). C, IHC staining of VEGF (upper panel) and WB analyses (lower panel) show increased induction of VEGF in degenerative FJC tissue (G3/4) compared to NFJC tissues (G0). D-F, IHC results demonstrate substantially increased neuro-filament-M (NF-M; a marker of neurite formation), neuronal growth promoting factor, NGF, and its cognate receptor TrkA in degenerative FJC (G3/4) compared to normal FJC tissues (G0). These results represent at least n = 3 for normal FJC tissue and n = 6 for degenerative FJC tissue (G1∼4). Osteoarthritis and Cartilage 2015 23, 2242-2251DOI: (10.1016/j.joca.2015.06.009) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Cytokine profiling in normal and degenerative facet joint capsular (FJC) tissues. A, Densitometric Analyses of Cytokine Antibody Array, using FJC tissue lysates from normal (G0) and degenerative FJ tissues (G3/4). The histogram shows levels of altered cytokines in degenerative FJC tissues (n = 10) relative to normal FJC tissues (n = 3). B, Quantitative real-time PCR analyses for comparative gene expression profile of pro- and anti-inflammatory cytokines and TLRs, between normal FJC tissues (G0) (NFJC, n = 3) and degenerative FJC tissues (DFJC, G3/4) (n = 10). Individual P-values represented in the figures. Data represented as mean. Error bars represented as 95% CI. Osteoarthritis and Cartilage 2015 23, 2242-2251DOI: (10.1016/j.joca.2015.06.009) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Expression of cartilage degrading enzymes and TIMPs in normal and degenerative FJC tissues. Comparison of MMPs, ADAMTSs, and TIMPs assessed by quantitative real-time PCR for mRNA levels (A-C, n = 3 normal (NFJC, G0); n = 12 degenerative FJC (DFJC, G3/4) tissue) (A–C) and WB for protein (n = 3 NFJC; n = 3 DFJC) (A′–C′). Individual P-values represented in the figures. Data represented as mean. Error bars represented as 95% CI. Osteoarthritis and Cartilage 2015 23, 2242-2251DOI: (10.1016/j.joca.2015.06.009) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 The level of inflammatory pain mediators and pain neuromodulators in normal and degenerative FJC tissues. Comparison of A, COX-2, iNOS; B, prostaglandin (EP) receptors; and C, pain neuromodulator (CGRP, Substance P, TRPV1, NGF, TrKA) expression in degenerative FJC tissue (G3/4, DFJC) as compared to normal FJC tissue (G0, NFJC), which was assessed by quantitative real-time PCR for mRNA (n = 3, NFJC; n = 10 DFJC tissue) and WB for protein (n = 3 NFJC tissue; n = 3 DFJC tissue). D, isolated rat DRGs were co-cultured with media control (M), NFJC tissue, or DFJC tissue for 5 days. Assessment of total relative gene induction was assessed by quantitative real-time PCR analyses of substance P and NGF in DRGs. The data represent three independent experiments (n = 3 donors) with triplicates. Individual P-values represented in the figures. Data represented as mean. Error bars represented as 95% CI. Osteoarthritis and Cartilage 2015 23, 2242-2251DOI: (10.1016/j.joca.2015.06.009) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions