Oliver Hausmann

Curcuma DMSO extracts and curcumin exhibit an anti-inflammatory and anti-catabolic effect on human intervertebral disc cells, possibly by influencing TLR2 expression and JNK activity.

BackgroundAs proinflammatory cytokines seem to play a role in discogenic back pain, substances exhibiting anti-inflammatory effects on intervertebral disc cells may be used as minimal-invasive therapeutics for intradiscal/epidural injection. The purpose of this study was to investigate the anti-inflammatory and anti-catabolic potential of curcuma, which has been used in the Indian Ayurvedic medicine to treat multiple ailments for a long time.MethodsHuman disc cells were treated with IL-1β to induce an inflammatory/catabolic cascade. Different extracts of curcuma as well as curcumin (= a component selected based on results with curcuma extracts and HPLC/MS analysis) were tested for their ability to reduce mRNA expression of proinflammatory cytokines and matrix degrading enzymes after 6 hours (real-time RT-PCR), followed by analysis of typical inflammatory signaling mechanisms such as NF-κB (Western Blot, Transcription Factor Assay), MAP kinases (Western Blot) and Toll-like receptors (real-time RT-PCR). Quantitative data was statistically analyzed using a Mann Whitney U test with a significance level of p < 0.05 (two-tailed).ResultsResults indicate that the curcuma DMSO extract significantly reduced levels of IL-6, MMP1, MMP3 and MMP13. The DMSO-soluble component curcumin, whose occurrence within the DMSO extract was verified by HPLC/MS, reduced levels of IL-1β, IL-6, IL-8, MMP1, MMP3 and MMP13 and both caused an up-regulation of TNF-α. Pathway analysis indicated that curcumin did not show involvement of NF-κB, but down-regulated TLR2 expression and inhibited the MAP kinase JNK while activating p38 and ERK.ConclusionsBased on its anti-inflammatory and anti-catabolic effects, intradiscal injection of curcumin may be an attractive treatment alternative. However, whether the anti-inflammatory properties in vitro lead to analgesia in vivo will need to be confirmed in an appropriate animal model.

Expression and regulation of toll-like receptors (TLRs) in human intervertebral disc cells.

PurposeAlthough inflammatory processes play an essential role in painful intervertebral disc (IVD) degeneration, the underlying regulatory mechanisms are not well understood. This study was designed to investigate the expression, regulation and importance of specific toll-like receptors (TLRs)—which have been shown to play an essential role e.g. in osteoarthritis—during degenerative disc disease.MethodsThe expression of TLRs in human IVDs was measured in isolated cells as well as in normal or degenerated IVD tissue. The role of IL-1β or TNF-α in regulating TLRs (expression/activation) as well as in regulating activity of down-stream pathways (NF-κB) and expression of inflammation-related genes (IL-6, IL-8, HSP60, HSP70, HMGB1) was analyzed.ResultsExpression of TLR1/2/3/4/5/6/9/10 was detected in isolated human IVD cells, with TLR1/2/4/6 being dependent on the degree of IVD degeneration. Stimulation with IL-1β or TNF-α moderately increased TLR1/TLR4 mRNA expression (TNF-α only), and strongly increased TLR2 mRNA expression (IL-1β/TNF-α), with the latter being confirmed on the protein level. Stimulation with IL-1β, TNF-α or Pam3CSK4 (a TLR2-ligand) stimulated IL-6 and IL-8, which was inhibited by a TLR2 neutralizing antibody for Pam3CSK4; IL-1β and TNF-α caused NF-κB activation. HSP60, HSP70 and HMGB1 did not increase IL-6 or IL-8 and were not regulated by IL-1β/TNF-α.ConclusionWe provide evidence that several TLRs are expressed in human IVD cells, with TLR2 possibly playing the most crucial role. As TLRs mediate catabolic and inflammatory processes, increased levels of TLRs may lead to aggravated disc degeneration, chronic inflammation and pain development. Especially with the identification of more endogenous TLR ligands, targeting these receptors may hold therapeutic promise.

Transcranial electrical stimulation: significance of fast versus slow charge delivery for intra-operative monitoring

OBJECTIVES Motor-evoked potentials (MEP) for intra-operative monitoring due to fast charge (fc: 1.0 Coulomb/s) and slow charge (sc: 0.1Coulomb/s) delivery for multipulse transcranial electrical stimulation (TES) were compared. METHODS MEPs due to fc (n=162) and sc stimulation (n=182) were performed in parallel in a prospective study. The fc stimulation technique is characterized by an increased steepness of charge delivery with consequent reduction of stimulus duration of 50 micros compared to 500 micros in sc stimulation. Stimulation charges (C=Coulomb) and MEP parameters during spine surgery were analyzed. RESULTS MEPs were successfully recorded in 15/18 patients under total intravenous anesthesia. The mean charge to induce intra-operative MEPs (stimulation threshold) was significantly less in fc (0.195 mC) as compared to sc stimulation (0.298 mC). With both stimulation techniques, in all patients without impairment of motor function, MEPs could be recorded and no technique was superior with respect to successful stimulation. The mean MEP latencies, amplitudes and the extent of intra-individual variation of MEP parameters during surgery (shift of latency less than 10%, variability of amplitude less than 50%) were not different with both stimulation techniques. CONCLUSIONS TES with either fc or sc stimulation can be used reliably for intraoperative monitoring. Fc and sc stimulation are comparable with respect to feasibility, intra-individual variability and mean parameters of MEP responses. However, fc stimulation provides a higher stimulation efficiency and requires about 35% less total charge for MEP monitoring.

SSEP analysis in surgery of idiopathic scoliosis: the influence of spine deformity and surgical approach.

Abstract. The study was conducted to assess the possible impact of spine deformity in patients with idiopathic scoliosis (IS) on tibial nerve somatosensory evoked potentials (t-SSEPs) and the influence of spine correction upon postoperative SSEP recordings. In 61 consecutive patients undergoing 64 spinal instrumentations, 129 pre- and postoperative SSEPs were analyzed. The degree of spine deformity was assessed by the pre-operative Cobb angle of the major scoliotic curve. In a control group, reference values of t-SSEP latencies were established with respect to body height. In a cohort study, IS patients were compared with healthy controls with respect to t-SSEP latency, amplitude, configuration and interside difference. The results of the analysis showed that preoperative-body-height-corrected t-SSEP latencies were prolonged in 61% of patients, with a pathological interside difference in 23.4% of them. The impairment of t-SSEPs was not related to the extent of spine deformity as assessed by the Cobb angle. Even without occurrence of postoperative neurological deficits, postoperative t-SSEPs showed significantly increased latencies without changes in t-SSEP configuration. The prolongation of t-SSEP latencies was related to the surgical procedure (combined ventro-dorsal approach), but not to the extent of spine correction, level of instrumentation, or number of fused segments. The analysis of preoperative t-SSEPs was of no predictive value for intra- or postoperative neurological complications. t-SSEPs are significantly affected in IS patients, although these patients show no obvious clinical neurological deficits. The extent of t-SSEP impairment is not related to the severity of scoliosis. Even in clinically uneventful surgery, the postoperative t-SSEPs can be deteriorated depending on the surgical approach. This indicates a subclinical impact of spine surgery upon spinal cord function.

Inflammaging in the intervertebral disc

Degeneration of the intervertebral disc – triggered by ageing, mechanical stress, traumatic injury, infection, inflammation and other factors – has a significant role in the development of low back pain. Back pain not only has a high prevalence, but also a major socio-economic impact. With the ageing population, its occurrence and costs are expected to grow even more in the future. Disc degeneration is characterized by matrix breakdown, loss in proteoglycans and thus water content, disc height loss and an increase in inflammatory molecules. The accumulation of cytokines, such as interleukin (IL)-1β, IL-8 or tumor necrosis factor (TNF)-α, together with age-related immune deficiency, leads to the so-called inflammaging – low-grade, chronic inflammation with a crucial role in pain development. Despite the relevance of these molecular processes, current therapies target symptoms, but not underlying causes. This review describes the biological and biomechanical changes that occur in a degenerated disc, discusses the connection between disc degeneration and inflammaging, highlights factors that enhance the inflammatory processes in disc pathologies and suggests future research avenues.

p38 MAPK Facilitates Crosstalk Between Endoplasmic Reticulum Stress and IL-6 Release in the Intervertebral Disc

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.