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Papers by Abbie Binch

Introduction This review will discuss the regulatory mechanisms of both innervation and vasculari... more Introduction This review will discuss the regulatory mechanisms of both innervation and vascularisation within normally aneural and avascular tissues, and how they may become altered in degeneration enabling new nerve and blood vessel formation which is hypothesised to be a source of pain. Conclusion Normal intervertebral discs and articular cartilage are the largest aneural and avascular tissues in the human body yet during intervertebral disc degeneration and osteoarthritis these tissues become increasingly vascularised by small blood vessels and innervated by peptide containing sensory nerve fibres. The mechanism by which this process occurs remains largely unknown. Published data suggests that various factors present within the healthy tissues such as aggrecan, chondromodulin and semaphorins may act as repulsive barriers to neurite and endothelial cell invasion. During degeneration however, the synthesis of these molecules becomes disrupted, potentially leading to vascularisatio...

Biomacromolecules, 2021

Adipose-derived mesenchymal stem cells (ASCs) have been identified for their promising therapeuti... more Adipose-derived mesenchymal stem cells (ASCs) have been identified for their promising therapeutic potential to regenerate and repopulate the degenerate intervertebral disk (IVD), which is a major cause of lower back pain. The optimal cell delivery system remains elusive but encapsulation of cells within scaffolds is likely to offer a decisive advantage over the delivery of cells in solution by ensuring successful retention within the tissue. Herein, we evaluate the use of a fully synthetic, thermoresponsive poly(glycerol monomethacrylate)-poly(2-hydroxypropyl methacrylate) (PGMA-PHPMA) diblock copolymer worm gel that mimics the structure of hydrophilic glycosaminoglycans. The objective was to use this gel to direct differentiation of human ASCs toward a nucleus pulposus (NP) phenotype, with or without the addition of discogenic growth factors TGFβ or GDF6. Accordingly, human ASCs were incorporated into a cold, free-flowing aqueous dispersion of the diblock copolymer, gelation induced by warming to 37 °C and cell culture was conducted for 14 days with or without such growth factors to assess the expression of characteristic NP markers compared to those produced when using collagen gels. In principle, the shear-thinning nature of the biocompatible worm gel enables encapsulated human ASCs to be injected into the IVD using a 21G needle. Moreover, we find significantly higher gene expression levels of ACAN, SOX-9, KRT8, and KR18 for ASCs encapsulated within worm gels compared to collagen scaffolds, regardless of the growth factors employed. In summary, such wholly synthetic worm gels offer considerable potential as an injectable cell delivery scaffold for the treatment of degenerate disk disease by promoting the transition of ASCs toward an NP-phenotype.

Nature Reviews Rheumatology, 2021

Intervertebral disc (IVD) degeneration is a major cause of low back pain, a prevalent and chronic... more Intervertebral disc (IVD) degeneration is a major cause of low back pain, a prevalent and chronic condition that has a striking effect on quality of life. Currently, no approved pharmacological interventions or therapies are available that prevent the progressive destruction of the IVD; however, regenerative strategies are emerging that aim to modify the disease. Progress has been made in defining promising new treatments for disc disease, but considerable challenges remain along the entire translational spectrum, from understanding disease mechanism to useful interpretation of clinical trials, which make it difficult to achieve a unified understanding. These challenges include: an incomplete appreciation of the mechanisms of disc degeneration; a lack of standardized approaches in preclinical testing; in the context of cell therapy, a distinct lack of cohesion regarding the cell types being tested, the tissue source, expansion conditions and dose; the absence of guidelines regarding disease classification and patient stratification for clinical trial inclusion; and an incomplete understanding of the mechanisms underpinning therapeutic responses to cell delivery. This Review discusses current approaches to disc regeneration, with a particular focus on cell-based therapeutic strategies, including ongoing challenges, and attempts to provide a framework to interpret current data and guide future investigational studies. Intervertebral disc (IVD) degeneration is a common condition associated with low back pain, but approved pharmacological interventions to prevent progression of IVD degeneration are lacking. This Review discusses the clinical progress and challenges of cell-based therapeutics for repairing the IVD. Intervertebral disc degeneration is a notable contributing factor to the incidence of low back pain. No pharmacological intervention, biologic therapy or procedure is approved for the prevention of disc degeneration; cell-based therapies are one approach currently being explored for promoting disc regeneration. Clinical trials investigating cell-based therapies are underway but are often poorly designed, with low patient numbers and an absence of appropriate controls. The broad range of disparate interventions relating to cell type, tissue source, expansion conditions, dose and delivery systems make it difficult to achieve unambiguous interpretation of results. Better imaging and biochemical diagnostics are needed to inform patient stratification protocols to identify cohorts of patients who would most benefit from cell therapy. Potential mechanisms of action of cell therapies have been proposed, including secretion of bioactive molecules, apoptosis and transfer of extracellular vesicles; however, concrete mechanistic evidence is lacking. Intervertebral disc degeneration is a notable contributing factor to the incidence of low back pain. No pharmacological intervention, biologic therapy or procedure is approved for the prevention of disc degeneration; cell-based therapies are one approach currently being explored for promoting disc regeneration. Clinical trials investigating cell-based therapies are underway but are often poorly designed, with low patient numbers and an absence of appropriate controls. The broad range of disparate interventions relating to cell type, tissue source, expansion conditions, dose and delivery systems make it difficult to achieve unambiguous interpretation of results. Better imaging and biochemical diagnostics are needed to inform patient stratification protocols to identify cohorts of patients who would most benefit from cell therapy. Potential mechanisms of action of cell therapies have been proposed, including secretion of bioactive molecules, apoptosis and transfer of extracellular vesicles; however, concrete mechanistic evidence is lacking.

JOR SPINE, 2020

Immunohistochemical analysis of protein expression in formalin fixed paraffin embedded human inte... more Immunohistochemical analysis of protein expression in formalin fixed paraffin embedded human intervertebral disc tissues. JOR SPINE, e1098.

Nature Communications, 2020

The ability to control nanostructure shape and dimensions presents opportunities to design materi... more The ability to control nanostructure shape and dimensions presents opportunities to design materials in which their macroscopic properties are dependent upon the nature of the nanoparticle. Although particle morphology has been recognized as a crucial parameter, the exploitation of the potential shape-dependent properties has, to date, been limited. Herein, we demonstrate that nanoparticle shape is a critical consideration in the determination of nanocomposite hydrogel properties. Using translationally relevant calcium-alginate hydrogels, we show that the use of poly(L-lactide)-based nanoparticles with platelet morphology as an adhesive results in a significant enhancement of adhesion over nanoparticle glues comprised of spherical or cylindrical micelles. Furthermore, gel nanocomposites containing platelets showed an enhanced resistance to breaking under strain compared to their spherical and cylindrical counterparts. This study opens the doors to a change in direction in the field ...

JOR SPINE, 2019

Background: Mesenchymal stem cells (MSCs) are becoming an increasingly attractive option for rege... more Background: Mesenchymal stem cells (MSCs) are becoming an increasingly attractive option for regenerative therapies due to their availability, self-renewal capacity, multilineage potential, and anti-inflammatory properties. Clinical trials are underway to test the efficacy of stem cell-based therapies for the repair and regeneration of the degenerate intervertebral disc (IVD), a major cause of back pain. Recently, both bone marrow-derived MSCs and adipose-derived stem cells (ASCs) have been assessed for IVD therapy but there is a lack of knowledge surrounding the optimal cell source and the response of transplanted cells to the low oxygen, pro-inflammatory niche of the degenerate disc. Here, we investigated several neurovascular factors from donormatched MSCs and ASCs that may potentiate the survival and persistence of sensory nerve fibers and blood vessels present within painful degenerate discs and their regulation by oxygen tensions and inflammatory cytokines. Methods: Donor-matched ASCs and MSCs were conditioned with either IL-1β or TNFα under normoxic (21% O 2) or hypoxic (5% O 2) conditions. Expression and secretion of several potent neurovascular factors were assessed using qRT-PCR and human magnetic Luminex assay. Results: ASCs and MSCs expressed constitutive levels of key neurotrophic factors; and stimulation of ASCs with hypoxia triggered increased secretion of both angiogenic factors (Ang-2 and VEGF-A) and neurotrophic (NGF and NT-3) compared to MSCs. We also report increased transcriptional regulation of pain-associated neuropeptides in hypoxia stimulated ASCs compared to those in normoxic conditions.

Osteoarthritis and Cartilage, 2017

Journal of Orthopaedic Research, 2016

Author contributions: JD & AB performed all laboratory work, and initial data analysis, contribut... more Author contributions: JD & AB performed all laboratory work, and initial data analysis, contributed to study design and helped draft the manuscript. CLM conceived the study, participated in its design and coordination, secured funding and assisted in data and statistical analysis, and co-wrote the manuscript. All authors read and approved the final manuscript. ROLE OF THE FUNDING SOURCE The funding source had no involvement in study design, collection, analysis or interpretation of data, writing of the manuscript or in the decision to submit the manuscript for publication. CONFLICT OF INTERESTS The author(s) declare that they have no conflicts of interests.

Oncotarget, Jan 19, 2016

Progress in mesenchymal stem cell (MSC) based therapies for nucleus pulposus (NP) regeneration ar... more Progress in mesenchymal stem cell (MSC) based therapies for nucleus pulposus (NP) regeneration are hampered by a lack of understanding and consensus of the normal NP cell phenotype. Despite the recent consensus paper on NP markers, there is still a need to further validate proposed markers. This study aimed to determine whether an NP phenotypic profile could be identified within a large population of mature NP samples.qRT-PCR was conducted to assess mRNA expression of 13 genes within human non-degenerate articular chondrocytes (AC) (n=10) and NP cells extracted from patients across a spectrum of histological degeneration grades (n=71). qRT-PCR results were used to select NP marker candidates for protein expression analysis.Differential expression at mRNA between AC and non-degenerate NP cells was only observed for Paired Box Protein 1 (PAX1) and Forkhead box F1 (FOXF1). In contrast no other previously suggested markers displayed differential expression between non-degenerate NP and ...

Background: Osteoarthritis (OA) is a common disease characterized by cartilage degeneration and j... more Background: Osteoarthritis (OA) is a common disease characterized by cartilage degeneration and joint remodeling. The underlying molecular changes underpinning disease progression are incompletely understood, but can be characterized using recent advances in genomics technologies, as the relevant tissue is readily accessible at joint replacement surgery. Here we investigate genes and pathways that mark OA progression, combining genome-wide DNA methylation, RNA sequencing and quantitative proteomics in isolated primary chondrocytes from matched intact and degraded articular cartilage samples across twelve patients with OA undergoing knee replacement surgery. Results: We identify 49 genes differentially regulated between intact and degraded cartilage at multiple omics levels, 16 of which have not previously been implicated in OA progression. Using independent replication datasets, we replicate statistically significant signals and show that the direction of change is consistent for ov...

Matrix Biology, 2016

The ECM of the intervertebral disc and articular cartilage contain a highly organised network of ... more The ECM of the intervertebral disc and articular cartilage contain a highly organised network of collagens and proteoglycans which resist compressive forces applied to these tissues. A pathological hallmark of the intervertebral disc is the imbalance between production of anabolic and catabolic factors by the resident cells. This process is thought to be mediated by proinflammatory cytokines, predominantly TNF-α and IL-1β, which upregulate expression of matrix degrading enzymes such as MMPs and ADAMTSs. This imbalance ultimately results in tissue degeneration causing failure of the biomechanical function of the tissues. A similar cascade of events is thought to occur in articular cartilage during development of osteoarthritis. Within these skeletal tissues a small, cell surface heparan sulphate proteoglycan; syndecan-4 (SDC4) has been implicated in maintaining physiological functions. However in the degenerating niche of the intervertebral disc and cartilage, dysregulated activities of this molecule may exacerbate pathological changes. Studies in recent years have elucidated a role for SDC4 in mediating matrix degradation in both intervertebral discs and cartilage by controlling ADAMTS-5 function and MMP3 expression. Discourse presented in this review highlights the potential of SDC4 as possible therapeutic target in slowing the progression of ECM degradation in both degenerative disc disease and osteoarthritis.

Oncotarget, 2015

Nerve and blood vessel ingrowth during intervertebral disc degeneration, is thought to be a major... more Nerve and blood vessel ingrowth during intervertebral disc degeneration, is thought to be a major cause of low back pain, however the regulation of this process is poorly understood. Here, we investigated the expression and regulation of a subclass of axonal guidance molecules known as the class 3 semaphorins, and their receptors; plexins and neuropilins within human NP tissue and their regulation by pro-inflammatory cytokines. Importantly this determined whether semaphorin expression was associated with the presence of nerves and blood vessels in tissues from human intervertebral discs. The study demonstrated that semaphorin3A, 3C, 3D, 3E and 3F and their receptors were expressed by native NP cells and further demonstrated their expression was regulated by IL-1β but to a lesser extent by IL-6 and TNFα. This is the first study to identify sema3C, sema3D and their receptors within the nucleus pulposus of intervertebral discs. Immunopositivity shows significant increases in semaphorin3C, 3D and their receptor neuropilin-2 in degenerate samples which were shown to contain nerves and blood vessels, compared to non-degenerate samples without nerves and blood vessels. Therefore data presented here suggests that semaphorin3C may have a role in promoting innervation and vascularisation during degeneration, which may go on to cause low back pain.

Global Spine Journal, 2014

Introduction Aquaporins are a family of transmembrane water channels that aid in osmoregulation, ... more Introduction Aquaporins are a family of transmembrane water channels that aid in osmoregulation, a critical function in the proteoglycan-rich nucleus pulposus. Previous reports have demonstrated expression of aquaporins in the intervertebral disc.1,2 Hypoxia, a defining feature of the nucleus pulposus environment has been shown to regulate expression of aquaporin1 (AQP1) and aquaporin 5 (AQP5).3-6 In some cases, this regulation has been shown to occur through HIF-1α and to depend on hypoxia responsive elements (HREs). The goal of this study was to examine if hypoxia and HIF-1α play a role in regulation and function of aquaporin1 (AQP1) and aquaporin5 (AQP5) in nucleus pulposus (NP) cells of the intervertebral disc. Materials and Methods Quantitative reverse transcription-polymerase chain reaction, Western blot, and immunohistochemistry were used to measure AQP1 and AQP5 expression in nucleus pulposus (NP) and annulus fibrosus (AF) cells and tissues. Transfections were used to determine the role of HIF-1α on AQP1 and AQP5 promoter activity in normoxia and hypoxia. The JASPAR database was used to identify two putative hypoxia response elements (HREs) in the promoter of each aquaporin. HIF-1α levels were modulated with treatment by DMOG or shRNAs. Expression of aquaporins in human tissue samples was evaluated using immunohistochemistry and polymerase chain reaction. Results We found that both AQP1 and AQP5 were expressed in tissues and cells of human and rat discs. To determine whether expression of the aquaporins was regulated by hypoxia, we treated NP cells with hypoxia (1% O2) for 8 to 72 hours. Treatment had no effect on promoter activity, mRNA expression, or protein expression of either aquaporin. Using the JASPAR database, we identified two HREs in the promoter of each aquaporin. Mutation of the HREs in either AQP1 or AQO5 had no suppressive effect on the basal activity of the promoters. We then investigated whether AQP1 and AQP5 were regulated in a HIF1α-dependent manner. Accumulation of HIF-1α by DMOG did not affect expression of either aquaporin. However, suppression of HIF-1α by lentiviral delivery of shRNA significantly decreased both mRNA and protein expression of AQP1 and AQP5. In addition, we have found changes in expression of these aquaporins in human tissue samples from patients with varying degrees of intervertebral disc degeneration. Conclusion These results indicate that, under hypoxic conditions, HIF-1α maintains basal expression of both AQP1 and AQP5 in the NP; however, this regulation is independent of HIF binding to the identified HREs in their promoters. Analysis of human tissue samples suggests that aquaporin expression may be linked to health of the intervertebral disc. Disclosure of Interest None declared References Richardson SM, Knowles R, Marples D, Hoyland JA, Mobasheri A. Aquaporin expression in the human intervertebral disc. J Mol Histol 2008;39(3):303–309 Gajghate S, Hiyama A, Shah M, et al. Osmolarity and intracellular calcium regulate aquaporin2 expression through TonEBP in nucleus pulposus cells of the intervertebral disc. J Bone Miner Res 2009;24(6):992–1001 Tanaka A, Sakurai K, Kaneko K, et al. The role of the hypoxia-inducible factor 1 binding site in the induction of aquaporin-1 mRNA expression by hypoxia. DNA Cell Biol 2011;30(8):539–544 Tie L, Lu N, Pan XY, et al. Hypoxia-induced up-regulation of aquaporin-1 protein in prostate cancer cells in a p38-dependent manner. Cell Physiol Biochem 2012;29(1-2):269–280 Zhang J, Xiong Y, Lu LX, et al. AQP1 expression alterations affect morphology and water transport in Schwann cells and hypoxia-induced up-regulation of AQP1 occurs in a HIF-1α-dependent manner. Neuroscience 2013;252(252):68–79 Kawedia JD, Yang F, Sartor MA, Gozal D, Czyzyk-Krzeska M, Menon AG. Hypoxia and hypoxia mimetics decrease aquaporin 5 (AQP5) expression through both hypoxia inducible factor-1α and proteasome-mediated pathways. PLoS ONE 2013;8(3):e57541

Global Spine Journal, 2014

Oncotarget, Jan 20, 2015

Objectives of this study were to investigate whether AQP1 and AQP5 expression is altered during i... more Objectives of this study were to investigate whether AQP1 and AQP5 expression is altered during intervertebral disc degeneration and if hypoxia and HIF-1 regulate their expression in NP cells. AQP expression was measured in human tissues from different degenerative grades; regulation by hypoxia and HIF-1 was studied using promoter analysis and gain- and loss-of-function experiments. We show that both AQPs are expressed in the disc and that mRNA and protein levels decline with human disease severity. Bioinformatic analyses of AQP promoters showed multiple evolutionarily conserved HREs. Surprisingly, hypoxia failed to induce promoter activity or expression of either AQP. While genomic chromatin immunoprecipitation showed limited binding of HIF-1α to conserved HREs, their mutation did not suppress promoter activities. Stable HIF-1α suppression significantly decreased mRNA and protein levels of both AQPs, but HIF-1α failed to induce AQP levels following accumulation. Together, our resul...

Journal of neurosurgical sciences, 2015

This review paper discusses the process of disc degeneration and the current understanding of cel... more This review paper discusses the process of disc degeneration and the current understanding of cellular degradation in patients who present with low back pain. The role of surgical treatment for low back pain is analysed with emphasis on the proven value of spinal fusion. The interesting and novel developments of stem cell research in the treatment of low back pain are presented with special emphasis on the importance of the cartilaginous end plate and the role of IL-1 in future treatment modalities.

Summary Anabolic and catabolic signalling processes within IVDs display overlapping pathways, how... more Summary Anabolic and catabolic signalling processes within IVDs display overlapping pathways, however some pathways were identified as selective to catabolic signalling and inhibition of one of these pathways inhibited some of the catabolic factors induced by IL-1 although NFkB inhibition also affected anabolic expression. Degeneration of intervertebral discs (IVDs) is implicated in 40% of low back pain cases. In the normal disc the balance between anabolic and catabolic processes are carefully balanced. During degeneration this balance is lost in favour of catabolic processes which lead to degradation of the IVD, infiltration of blood vessels and nerves and release of cytokines which sensitise nerves to pain. Interleukin 1 (IL-1) is known to be important in the pathogenesis of IVD degeneration, here we investigated the intracellular signalling pathways activated by IL-1 and those activated by an anabolic factor (CDMP-1) to investigate differential pathways. Human nucleus pulposus cells (NP) removed during discetomy for nerve root pain were stimulated with IL-1 or CDMP-1 for 30 minutes. Site-specific phosphorylation of 46 signalling molecules were identified using R&D proteome array. The activation of ERK1/2, p38, c-jun, and IkB were confirmed using cell based ELISAs, in addition pNFκB localisation in stimulated cells was determined using immunohistochemisty. Pre-treatment with inhibitors to p38, and NFkB for 30 minutes, followed by stimulation with IL-1 (10ng/mL) or CDMP-1 (10ng/mL) for 24 hours was investigated to determine effects on anabolic and catabolic factors. In addition localisation of phosphorylated c-jun, p38 and NFkB were investigated within paraffin embedded sections of human IVD to investigate the presence of active pathways in vivo . Twenty intracellular signalling pathways were activated following CDMP-1 treatment and 8 signalling pathways activated by IL-1. Of note key classical IL-1 signalling pathways p38 MAPK, ERK 1/2 and JNK were activated by IL-1, however of these ERK 1/2 particularly was also activated by CDMP-1, whilst p38 and c-jun were only activated by IL-1. IL-1 induced activation of NFkB signalling to a greater extent than CDMP-1, these results were confirmed by the ‘in cell ELISAs’. IVD tissue samples displayed immunopositive staining for phosphorylated c-jun, NFkB and p38. Inhibition of p38 signalling inhibited IL-1 induced MMP 13 expression, but had little effect on the induction of IL-8. However inhibitors of NFkB signalling pathway failed to inhibit the induction of MMP 13 but abrogated the induced IL-6 and IL-8 expression. IL-1 induced a complete aberration of aggrecan expression by NP cells in alginate culture, this effect was partly inhibited by p38 MAPK inhibitor but was completely restored by inhibiting NFkB signalling. However the aggrecan expressed in CDMP-1 treated cells was decreased by inhibiting NFkB but not p38. Here, we have shown that anabolic and catabolic signalling processes within IVDs show a number of overlapping pathways, however a number of differential pathways were identified and inhibition of p38 MAPK and NFkB pathways inhibited a number of catabolic processes investigated which were induced by IL-1. Thus inhibition of signalling pathways could be a novel mechanism of inhibiting catabolic processes which could hold promise to inhibit degeneration at early stages of disease but also create the correct tissue niche to promote regeneration of the disc.

Summary Cytokines produced within the degenerate disc induce expression of neurotrophic factors a... more Summary Cytokines produced within the degenerate disc induce expression of neurotrophic factors and pain related peptides which could be important in nerve ingrowth and pain sensitisation leading to low back pain. The intervertebral disc (IVD) is considered the largest aneural and avascular structure within the human body, yet during degeneration vascularisation of the IVD is seen to be accompanied by nociceptive nerves. Low back pain is a highly debilitating condition affecting around 80% of the population, 40% of which are attributed to IVD degeneration. Discogenic pain was largely thought to be a result of irritation and compression of the nerve root, yet recent data suggests that pain may be attributed to the sensitisation of sensory nerves by the synthesis of pain related peptides, calcitonin gene related peptide (CGRP) and substance P. It is known that cytokines and chemokines produced by nucleus pulposus cells elicit various effects including the production of matrix degrading enzymes, and decreased matrix molecules. Here, we investigate the hypothesis that cytokines regulate both neurotrophic factor and pain related peptide synthesis within nucleus pulposus and nerve cells which may elicit algesic effects. Real-Time PCR was performed to investigate gene expression of the neurotrophic factors NGF, BDNF, NT3 and their receptors Trk A, B and C along with Substance P and CGRP on directly extracted RNA from human NP cells and NP cells cultured in alginate for 2 weeks prior to treatment for 48hours with IL-1, IL-6 or TNFα at 0–100ng/mL. Similarly SH-SY5Y neuroblastoma cells were differentiated in retinoic acid for 7 days prior to stimulation with IL-1, IL-6 or TNFα at 0ng/mL and 10ng/mL for 48hours. Immunohistochemistry was used to localise neurotrophic factor receptors Trk A, B and C in both degenerate discs and neuronal cells. NGF expression was present in normal and degenerate disc samples, however only degenerate discs expressed the high affinity receptor TrkA. Similarly Trk B was present in 22% of normal samples increasing to 100% expression within degenerate disc samples. All cytokines increased expression of NGF in NP cells (P≤0.05). TNFα also increased BDNF significantly, whereas no significant affects were seen in NT3 expression in NP cells. Trk B expression was significantly increased by IL-1 and TNFα treatment of NP cells. Conversely Trk C was down regulated by IL-6. Substance P was significantly increased by IL-1 and TNFα treatments whilst IL-6 and TNFα increased CGRP expression in NP cells. In SH-SY5Y cells, IL-1 significantly increased BDNF whilst IL-6 and TNFα failed to induce significant differences in neurotrophic factors. All cytokines increased Trk expression in the nerve cell line; however this failed to reach significance. Immunohistochemistry confirmed the presence of Trk receptors within the neuronal cell line. Here we have demonstrated that a number of cytokines known to be up regulated during disc degeneration and disc prolapse, induce expression of various neurotrophic factors, their receptors and pain related peptides within human NP cells, as well as SH-SY5Y cells. This data suggests that the presence and production of cytokines within the degenerate disc may be responsible for nerve ingrowth and sensitisation of nerves which may result in discogenic pain.

Arthritis Research & Therapy, 2014

Introduction: The degenerate intervertebral disc (IVD) becomes innervated by sensory nerve fibres... more Introduction: The degenerate intervertebral disc (IVD) becomes innervated by sensory nerve fibres, and vascularised by blood vessels. This study aimed to identify neurotrophins, neuropeptides and angiogenic factors within native IVD tissue and to further investigate whether pro-inflammatory cytokines are involved in the regulation of expression levels within nucleus pulposus (NP) cells, nerve and endothelial cells. Methods: Quantitative real-time PCR (qRT-PCR) was performed on 53 human IVDs from 52 individuals to investigate native gene expression of neurotrophic factors and their receptors, neuropeptides and angiogenic factors. The regulation of these factors by cytokines was investigated in NP cells in alginate culture, and nerve and endothelial cells in monolayer using RT-PCR and substance P (SP) protein expression in interleukin-1 (IL-1β) stimulated NP cells. Results: Initial investigation on uncultured NP cells identified expression of all neurotrophins by native NP cells, whilst the nerve growth factor (NGF) receptor was only identified in severely degenerate and infiltrated discs, and brain derived neurotrophic factor (BDNF) receptor expressed by more degenerate discs. BDNF expression was significantly increased in infiltrated and degenerate samples. SP and vascular endothelial growth factor (VEGF) were higher in infiltrated samples. In vitro stimulation by IL-1β induced NGF in NP cells. Neurotropin-3 was induced by tumour necrosis factor alpha in human dermal microvascular endothelial cells (HDMECs). SP gene and protein expression was increased in NP cells by IL-1β. Calcitonin gene related peptide was increased in SH-SY5Y cells upon cytokine stimulation. VEGF was induced by IL-1β and interleukin-6 in NP cells, whilst pleiotrophin was decreased by IL-1β. VEGF and pleiotrophin were expressed by SH-SY5Y cells, and VEGF by HDMECs, but were not modulated by cytokines. Conclusions: The release of cytokines, in particular IL-1β during IVD degeneration, induced significant increases in NGF and VEGF which could promote neuronal and vascular ingrowth. SP which is released into the matrix could potentially up regulate the production of matrix degrading enzymes and also sensitise nerves, resulting in nociceptive transmission and chronic low back pain. This suggests that IL-1β is a key regulatory cytokine, involved in the up regulation of factors involved in innervation and vascularisation of tissues.

Global Spine Journal, 2014

Biomacromolecules, 2021

Nature Reviews Rheumatology, 2021

JOR SPINE, 2020

Nature Communications, 2020

JOR SPINE, 2019

Osteoarthritis and Cartilage, 2017

Journal of Orthopaedic Research, 2016

Oncotarget, Jan 19, 2016

Matrix Biology, 2016

Oncotarget, 2015

Global Spine Journal, 2014

Oncotarget, Jan 20, 2015

Journal of neurosurgical sciences, 2015

Arthritis Research & Therapy, 2014

Global Spine Journal, 2014