Degenerative disc disease is associated with increased expression of pro-inflammatory cytokines in the intervertebral disc (IVD). However, it is not completely clear how inflammation arises in the IVD and which cellular compartments are involved in this process. Recently, the endoplasmic reticulum (ER) has emerged as a possible modulator of inflammation in age-related disorders. In addition, ER stress has been associated with the microenvironment of degenerated IVDs. Therefore, the aim of this study was to analyze the effects of ER stress on inflammatory responses in degenerated human IVDs and associated molecular mechanisms. Gene expression of ER stress marker GRP78 and pro-inflammatory cytokines IL-6, IL-8, IL-1β, and TNF-α was analyzed in human surgical IVD samples (n = 51, Pfirrmann grade 2–5). The expression of GRP78 positively correlated with the degeneration grade in lumbar IVDs and IL-6, but not with IL-1β and TNF-α. Another set of human surgical IVD samples (n = 25) was used to prepare primary cell cultures. ER stress inducer thapsigargin (Tg, 100 and 500 nM) activated gene and protein expression of IL-6 and induced phosphorylation of p38 MAPK. Both inhibition of p38 MAPK by SB203580 (10 µM) and knockdown of ER stress effector CCAAT-enhancer-binding protein homologous protein (CHOP) reduced gene and protein expression of IL-6 in Tg-treated cells. Furthermore, the effects of an inflammatory microenvironment on ER stress were tested. TNF-α (5 and 10 ng/mL) did not activate ER stress, while IL-1β (5 and 10 ng/mL) activated gene and protein expression of GRP78, but did not influence [Ca²⁺]_i flux and expression of CHOP, indicating that pro-inflammatory cytokines alone may not induce ER stress in vivo. This study showed that IL-6 release in the IVD can be initiated following ER stress and that ER stress mediates IL-6 release through p38 MAPK and CHOP. Therapeutic targeting of ER stress response may reduce the consequences of the harsh microenvironment in degenerated IVD.

Transient receptor potential (TRP) channels have emerged as potential sensors and transducers of inflammatory pain. The aims of this study were to investigate (1) the expression of TRP channels in intervertebral disc (IVD) cells in normal and inflammatory conditions and (2) the function of Transient receptor potential ankyrin 1 (TRPA1) and Transient receptor potential vanilloid 1 (TRPV1) in IVD inflammation and matrix homeostasis. RT-qPCR was used to analyze human fetal, healthy, and degenerated IVD tissues for the gene expression of TRPA1 and TRPV1. The primary IVD cell cultures were stimulated with either interleukin-1 beta (IL-1β) or tumor necrosis factor alpha (TNF-α) alone or in combination with TRPA1/V1 agonist allyl isothiocyanate (AITC, 3 and 10 µM), followed by analysis of calcium flux and the expression of inflammation mediators (RT-qPCR/ELISA) and matrix constituents (RT-qPCR). The matrix structure and composition in caudal motion segments from TRPA1 and TRPV1 wild-type (WT) and knock-out (KO) mice was visualized by FAST staining. Gene expression of other TRP channels (A1, C1, C3, C6, V1, V2, V4, V6, M2, M7, M8) was also tested in cytokine-treated cells. TRPA1 was expressed in fetal IVD cells, 20% of degenerated IVDs, but not in healthy mature IVDs. TRPA1 expression was not detectable in untreated cells and it increased upon cytokine treatment, while TRPV1 was expressed and concomitantly reduced. In inflamed IVD cells, 10 µM AITC activated calcium flux, induced gene expression of IL-8, and reduced disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) and collagen 1A1, possibly via upregulated TRPA1. TRPA1 KO in mice was associated with signs of degeneration in the nucleus pulposus and the vertebral growth plate, whereas TRPV1 KO did not show profound changes. Cytokine treatment also affected the gene expression of TRPV2 (increase), TRPV4 (increase), and TRPC6 (decrease). TRPA1 might be expressed in developing IVD, downregulated during its maturation, and upregulated again in degenerative disc disease, participating in matrix homeostasis. However, follow-up studies with larger sample sizes are needed to fully elucidate the role of TRPA1 and other TRP channels in degenerative disc disease.

European Cells & Materials Journal - Open and Free Access - The Official Research Journal of AOCMF, AOTrauma, European Orthopaedic Research Society (EORS), Swiss Society for Biomaterials (SSB) and Tissue & Cell Engineering Society (TCES)

Objectives: In chronic low back pain (CLBP) patients, study of altered brain metabolites in the anterior cingulate cortex (ACC) using magnetic resonance spectroscopy (MRS) could reveal the detailed pathology of CLBP and depression. The aim was to detect the central difference between CLBP and controls by means of measuring the metabolites in the ACC, and to analyze the correlations between depression and metabolites in ACC. Materials and Methods: MRS was performed in CLBP (n=60) and control participants (n=56) to evaluate the effects of CLBP on metabolites in the ACC and to analyze the correlations between metabolites and questionnaire scores in a cross-sectional study. Results: Adjusting for age and sex, a negative effect of CLBP on the N-acetylaspartate (NAA) level (estimated regression slope coefficient [B]=−0.685, P<0.001) and positive effects on the glutamate +glutamine (Glx)/creatine (B=0.136, P=0.016) and Glx/myoinositol (B=0.140, P0.4, P<0.05). Discussion: Lower NAA levels and higher Glx/creatine and Glx/myoinositol ratios in the ACC of CLBP participants compared with controls were revealed. The result suggests the hypothesis that excessive Glx leads to neuronal dysfunction and/or death, which was reflected as a low NAA level in the ACC of individuals with CLBP. Measurement of these metabolites using MRS potentially helps evaluate CLBP patients’ condition and psychological status objectively.

Recent studies report that surfaces displaying micrometer- or nanometer-sized undulating structures exhibit antibacterial effects. In previous work, we described the use of an advanced nanofabrication technique to generate an artificial biomimetic Moth-eye film by coating a polyethylene terephthalate (PET) film with nanoscale moth-eye protrusions made from a hydrophilic resin. This moth-eye film exhibited enhanced antibacterial effects in in vitro experiments. The aim of the present study was to verify the antibacterial efficacy of the Moth-eye film in practical environments. The antibacterial effects of three types of film (Moth-eye film, Flat film, and PET film) were compared. Sample films were pasted onto hand washing basins at the testing locations. After several hours, bacteria were collected from the surface of the sample films with one of three kinds of culture media stamper (to permit identification of bacterial species). The stampers were incubated for 48 hours at 35°C, and the numbers of colonies were counted. The number of common bacteria including E. coli and S. aureus obtained from the Moth-eye film was significantly lower than those from the PET film (p<0.05) and Flat film at 1 hour (p<0.05). This study found that the Moth-eye film showed a long-term (6h) antibacterial effect and the Moth-eye structure (PET coated with nanoscale cone-shaped pillars) demonstrated a physical antibacterial effect from earlier time points. Therefore, the Moth-eye film appears to have potential general-purpose applications in practical environments.

Beauty parlor stroke syndrome (BPSS) is a rare condition characterized by mechanical impingement of a vertebral artery (VA) during neck rotation and/or hyperextension followed by vertebrobasilar insufficiency. However, there have been no reports of BPSS in which the cause of mechanical impingement was identified and no cases for which surgical treatment was reported. The authors report the case of a 56-year-old Japanese man who presented with presyncope that occurred during cervical extension. Given the possibility of vertebrobasilar insufficiency, digital subtraction angiography and CT angiography were performed. These studies revealed that the right VA was hypoplastic and the left VA was dominant. Moreover, in the position of cervical extension, the dominant left VA showed constriction caused by a bone fragment of an osteophyte of the atlas. Removal of the bone fragment was performed. Postoperative left vertebral angiography showed improvement of blood flow in the extended position, and the presyncope completely disappeared. The pathomechanism of this case was a bone fragment compressing the left VA in the C-1 groove during neck extension. In BPSS patients with recurrent transient symptoms, the possibility of this mechanism of VA constriction by a free bone fragment should be considered.