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Music Factory (as known as The Music Factory Entertainment Group) is a UK promotional remix service that started in 1985. To this day, the service provides monthly Mastermix albums with exclusive mixes to qualified disc jockeys, as well as having provided a number of spinoff services over the years (some remixed, some not). It is also responsible for the Jive Bunny and the Mastermixers releases that were made commercially available.
Abstract:Previously, we proposed the hypothesis that similarities in the inflammatory response observed in acne vulgaris and degenerative disc disease (DDD), especially the central role of interleukin (IL)-1β, may be further evidence of the role of the anaerobic bacterium Cutibacterium (previously Propionibacterium) acnes in the underlying aetiology of disc degeneration. To investigate this, we examined the upregulation of IL-1β, and other known IL-1β-induced inflammatory markers and neurotrophic factors, from nucleus-pulposus-derived disc cells infected in vitro with C. acnes for up to 48 h. Upon infection, significant upregulation of IL-1β, alongside IL-6, IL-8, chemokine (C-C motif) ligand 3 (CCL3), chemokine (C-C motif) ligand 4 (CCL4), nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), was observed with cells isolated from the degenerative discs of eight patients versus non-infected controls. Expression levels did, however, depend on gene target, multiplicity and period of infection and, notably, donor response. Pre-treatment of cells with clindamycin prior to infection significantly reduced the production of pro-inflammatory mediators. This study confirms that C. acnes can stimulate the expression of IL-1β and other host molecules previously associated with pathological changes in disc tissue, including neo-innervation. While still controversial, the role of C. acnes in DDD remains biologically credible, and its ability to cause disease likely reflects a combination of factors, particularly individualised response to infection.Keywords: Cutibacterium acnes; disc cells; co-culture; inflammation; neurotrophic factors; gene expression; intracellular
Low back pain is a major cause of disability especially for people between 20 and 50 years of age. As a costly healthcare problem, it imposes a serious socio-economic burden. Current surgical therapies fail to replace the normal disc in facilitating spinal movements and absorbing load. The focus of regenerative medicine is on identifying biomarkers and signalling pathways to improve our understanding about cascades of disc degeneration and allow for the design of specific therapies. We hypothesized that comparing microarray profiles from degenerative and non-degenerative discs will lead to the identification of dysregulated signalling and pathophysiological targets. Microarray data sets were generated from human annulus fibrosus cells and analysed using IPA ingenuity pathway analysis. Gene expression values were validated by qRT-PCR and respective proteins were identified by immunohistochemistry. Microarray analysis revealed 238 differentially expressed genes in the degenerative annulus fibrosus. Seventeen of the dysregulated molecular markers showed log2-fold changes greater than ±1.5. Various dysregulated cellular functions, including cell proliferation and inflammatory response, were identified. The most significant canonical pathway induced in degenerative annulus fibrosus was found to be the interferon pathway. This study indicates interferon-alpha signalling pathway activation with IFIT3 and IGFBP3 up-regulation, which may affect cellular function in human degenerative disc.
Low back pain (LBP) is a major cause of disability that has become a serious socio-economic burden1. Although LBP is common in people who are between 20 and 50 years old, the burden is more persistent in the older population2. According to the WHO, LBP is the most costly healthcare problem and estimates suggest that the total costs exceed $100 billion per year in the United States alone3. LBP is commonly linked to the degeneration of the intervertebral disc (IVD)4,5. Discogenic low back pain is considered to originate from critical tears and fissures in the annulus fibrosus (AF) leading to nerve ingrowth and crucial weakness of the outer annulus that eventually results in a protrusion of nucleus pulposus (NP) tissue6. Annular defects may arise from trauma, aging or disc degeneration and may lead to extracellular matrix (ECM) degradation with concomitant loss of proteoglycan, tissue hydration and disc height. This weakens the AF such that it cannot withstand the hydrostatic pressures from the NP nor stabilise the functional spine unit7.
Current surgical therapy options include the removal of the degenerated or herniated tissue or even the partial or complete replacement of the disc with an artificial substitute8. However, these approaches have considerable drawbacks, such as the risk for adjacent disc degeneration and the failure of the artificial implants to accurately replace the normal disc in terms of movements and absorbing the load pressure9,10. Therapeutic intervention at an early stage of degeneration could avoid the need for such highly invasive procedures. Although substantial advancement has been achieved in identifying the multifactorial mechanisms that encompass the degenerative flow, knowledge about molecular pathways involved in its initiation and progression is still limited. Therefore, the current focus of regenerative medicine is on identifying biomarkers and signalling pathways that provide a better understanding of the cascades of disc degeneration.
The abnormal cell-mediated response, the changes that occur in the extracellular matrix composition11 and the diminished biomechanical characteristics12, which can be induced by non-physiological mechanical loading13, genetic predisposition and decreased cell activity14, lead to gradual structural failure of the IVD. This condition defines the degenerative disc disease (DDD) that goes along with nerve in-growth and low back pain15. Disc degeneration is also accompanied by inflammation, which is one of the major factors leading to phenotype changes and apoptosis. During disc degeneration, anabolic metabolism is decreased, whereas the catabolic molecular markers are increased16. Importantly, comparison between healthy and degenerated discs shows an imbalance of inflammatory cytokines that significantly increase during the degenerative process. Among the inflammatory cytokines discovered to date in disc degeneration and herniation in patients with severe LBP in comparison with healthy tissues, interleukin (IL)1-beta and tumour necrosis factor (TNF)-alpha are most prominent17. During the progression of the degenerative process, many other inflammatory cytokines and catabolic mediators such as prostaglandin E2, nitric oxide, IL6, IL8, matrix metalloproteases (MMPs), a disintegrin and metalloprotease with thrombospondin motif (ADAMTS)4 and ADAMTS5 enzymes and the death-inducing ligand Fas synchronize and consequently degrade the extracellular matrix18,19.
Recently, the focus has shifted towards identifying signalling pathways affecting the cellular and molecular functions and highlighting the underlying molecular markers for better understanding of the degenerative process in the disc. One of the most important pathways identified recently is the Wnt signalling that may mediate IVD degeneration by activation of MMPs and degradation of matrix molecules leading to NP cell senescence20. Moreover, caveolin-1, which regulates the Wnt signalling pathway, was reported to be up-regulated in the human degenerated IVD and was correlated with an increase in cell senescence markers21. On the other hand, Smolders et al. reported that in the dog NP, caveolin-1 and hence Wnt/β-catenin signalling pathway is crucial for the preservation of notochordal cells and hence for disc regeneration22. Furthermore, signalling pathways that regulate pro-inflammatory processes are activated during IVD degeneration and have been reported as potential therapeutic targets. Specifically, nuclear factor kappa B (NF-kB) and mitogen-activated protein kinase (MAPK) pathways have been identified as key regulators of inflammation, matrix catabolism and pain23. Recent findings also indicate a role of Notch signalling in the progress of disc degeneration, as Notch receptors and target genes were up-regulated in disc cells following treatment with pro-inflammatory cytokines24.
Nevertheless, further investigation into dysregulated processes in degenerative human disc is essential to provide more prospects for therapeutic targets. Large-scale assessment of molecular profiles enables us to comprehensively search for molecular markers and pathways associated with impaired cell functions in the human disc degenerative process. A genome-wide analysis of human AF samples was undertaken by Gruber et al. and focussed on the expression of genes associated with pain, neurotrophin and nerve regulation25. Significant changes in numerous genes related to these ontologies were found in more degenerated compared to less degenerated discs. In a similar study, microarray analysis was used to identify the expression patterns of genes related to mitochondrial function in human AF specimens, whereby the expression changes indicated mitochondrial dysfunction in degenerative AF26. Furthermore, gene expression profiling was utilised to detect differences between notochordal and chondrocyte-like nucleus pulposus cells from non-chondrodystrophic and chondrodystrophic dogs22; interestingly, dysregulation of canonical Wnt signalling was associated with early disc degeneration in chondrodystrophic breeds. Recently, several differentially expressed long noncoding RNAs were identified by microarray analysis of degenerative versus non-degenerative human NP samples, expanding our understanding of aberrant gene regulation in DDD27. 2b1af7f3a8