INTRODUCTION: Although it is well recognised that the outer annulus is innervated, the relative densities of innervation of different regions of the disc have not been quantitated. We present here the first comparative analysis of the innervation of the innervation of different regions of the lumbar intervertebral disc. METHODS: A sheep model was used allowing evaluation of the whole motion segment. Four sheep spines were used. One was processed for PGP 9.5 immunofluorescence and three were processed for PGP 9.5 immunoperoxidase histochemistry. Serial sagittal sections were obtained and a count was made of the densities of innervation of different regions of the endplate and annulus. These were compared to identify which areas of the disc and endplate are most innervated. RESULTS: The endplate innervation is concentrated centrally adjoining the nucleus. The mean density of innervation of the central endplate was 0.44 (SEM 0.07) nerves/mm2 while the mean density of the peripheral endplate was 0.10 (SEM 0.03) nerves/ mm. 2. (p= 0.0001). There was no significant difference between the overall endplate and annulus innervation densities 0.52 (SEM 0.1) v 0.37 (SEM 0.07) p=0.2. But the peri-annular connective tissue, external to the outer annulus contained the densest innervation of any region in the motion segment 1.05 (SEM 0.16). DISCUSSION: The lumbar intervertebral disc has a meagre innervation. This is concentrated in the peri-annular connective tissue and the central endplate. While receptor threshold is more closely related to noci-ceptive function than innervation density, these findings have important implications for any treatment of discogenic pain

Introduction: Although it is well recognized that the outer annulus is innervated, the relative densities of innervation of different regions of the disc have not been quantitated. We present here the first comparative analysis of the innervation of the innervation of different regions of the lumbar intervertebral disc. Methods: A sheep model was used allowing evaluation of the whole motion segment. Four sheep spines were used. One was processed for PGP 9.5 immunoflourescence and three were processed for PGP 9.5 immunoperoxidase histochemistry. Serial sagittal sections were obtained and a count was made of the densities of innervation of different regions of the endplate and annulus. These were compared to identify which areas of the disc and endplate are most innervated. Results: The endplate innervation is concentrated centrally adjoining the nucleus. The mean density of innervation of the central endplate was 0.44 (SEM 0.07) nerves/ mm. 2. while the mean density of the peripheral endplate was 0.10 (SEM 0.03) nerves/ mm. 2. (p= 0.0001). There was no significant difference between the overall endplate and annulus innervation densities 0.52 (SEM 0.1) v 0.37 (SEM 0.07) p=0.2. But the peri-annular connective tissue, external to the outer annulus contained the densest innervation of any region in the motion segment 1.05 (SEM 0.16). Discussion: The lumbar intervertebral disc has a meagre innervation. This is concentrated in the peri-annular connective tissue and the central endplate. While receptor threshold is more closely related to nociceptive function than innervation density, these findings have important implications for any treatment of discogenic pain

Background: The Inferior Glenohumeral Ligament (IGHL) has a well known mechanical and propioceptive relevance in shoulder stability. The interrelation of the IGHL anatomical disposition and innervation has not actually been described. The studys purpose was to determine the IGHL innervation patterns and relate them to dislocation. Material & methods: Forty-five embalmed and 16 fresh-frozen human cadaveric shoulders were studied. Massons Trichrome staining was used to detail the intra-ligamentous nerve fibre arrangements. Neural behaviour of the articular nerves was studied dynamically at the apprehension position and while anteroinferior dislocation of the shoulder joint was performed. Results: The anatomy of the IGHL was clearly defined. However, in 7 out of 61 cases the anterior band was only a slight thickening of the ligament. It averaged 34 mm (range, 28 to 46 mm) in length. The posterior band was only seen in 40.98 % of the cases. The axillary nerve provided IGHL innervation in 95.08 % of the cases. We found two distinct innervation patterns originating in the axillary nerve. In Type 1 (29.5 % of the cases), one or two collaterals later diverged from the main trunk to enter the ligament. Type 2 (65.57%) showed innervation to the ligament provided by the posterior branch for three to four neural branches. In both cases, these branches enter the ligament near the glenoid rim and at 7 oclock position (right shoulder). The shortest distance to the glenohumeral capsule was noted at 5 oclock position. The radial nerve (Type 3 innervation pattern) provided IGHL innervation in 3.28 % (2 specimens). Microscopic analysis revealed wavy intraligamentous neural branches. The articular branches relaxed and separated from the capsule at external rotation and abduction and stayed intact after dislocation. Conclusions: The current results showed the IGHL to have three different innervation patterns. The special neural anatomy of the IGHL suggested it was designed to avoiding denervation when dislocated. This might contribute to understand why the neural arch remains unaffected after most dislocations. To our knowledge this is the first work that clearly describes specimens in which the main innervation of the IGHL is provided by the radial nerve. Knowledge of the neural anatomy of the shoulder will clearly help in avoiding its injury in surgical procedures

Purpose of the study: The piriform syndrome is treated medically: functional rehabilitation and injections. If the medical treatment fails, tenotomy of the piriform muscle can be proposed. Published studies report good outcome in 66 to 87% Of patients. The purpose of this study was to examine the extrapelvic innervations of this muscle in order to assess the feasibility of neurotomy of he piriform muscle. Material and method: Twenty gluteal regions were dissected. We studied first the relations between the piriform muscle and the ischiatic nerve. Then the innervations branches of the piriform muscle were localized in three landmarks. Results: We found the of the six types of relation between the ischiatic nerve and the piriform muscle described by Beaton, with frequencies comparable to reports in the literature. Innervation of the piriform muscle does not follow a standard pattern, even though the innervations generally comes from the ischiatic nerve; the nerve branches come from the superior and inferior gluteal pedicles. In addition, these nerve branches penetrate the deep aspect of the muscle in random fashion. In addition, accessibility to the deep aspect of the piriform muscle cannot be achieved easily but requires prior section of its insertion on the greater trochanter. Discussion: In our opinion, these results suggest that isolated neurotomy of the piriform muscle is not clinically feasible; it might be possible to improve function results of isolated tenotomy by performing a neurotomy of the nerve branches visible during the tenotomy procedure. A greater benefit might be expected in forms where the ischiatic nerve crosses the piriform muscle

Purpose of the study: Spastic hypertony of the upper limb produces pronation of the forearm with flexion of the wrist and fingers. Treatment is generally based on injections of botulinum toxin and sometimes on selective neurotomy. Material and methods: In order to achieve better selection of the motor branches innervating the muscles requires a precise knowledge of the extramuscular innervation. Similarly, for botulinum toxin, injections must be made as close as possible to zones with the greatest density of intramuscular nerve endings, considered as the zones having the greatest number of neuromuscular junctions. Knowledge of these zones is currently insufficient. We therefore conducted a macroscopic then microscopic dissection of the muscles of the ventral forearm in 30 specimens to study extra- and intra-muscular innervations and the distributions of the nerve endings. Results: Surface maps were drawn to describe the precise localization of the motor branches for each muscle. These maps were designed as guides for surgical approaches for selective neurotomy. Then for each muscle, the zones with the greatest density of nerve endings were delimited in segments which could be used to define optimal zones of injection of botulinum toxin

Purpose of the study: Intramuscular injection of the botulinum toxin into the psoas can be proposed for permanent hip flexion due to spastic disorders. Several approaches have been described: retrograde subinguinal, anterolateral suprailiac, and posterior. Ultrasound or computed tomography can be used to guide needle position. These approaches are however limited to access to the L4 region, i.e. far from the motor points and with the risk of injury to the ureter. The purpose of this work was to determine the innervations of the psoas muscle that would be best adapted to this type of injection and thus to describe the most effective and reliable approach. Material and methods: This anatomy study included 20 dissections to: describe vertebral insertions of the psoas major and the psoas minor and to measure their distance from the iliac crest; define the region where the ureter crosses in front of the psoas. Results: More than 80% of the psoas muscles presented a proximal insertion on the transverse process of T12 and the body of L1; the mean length of the psoas in the adult is 27 cm above the inguinal ligament; the nerve roots collateral to the lumbar plexus are: 33% L2, 25% L3, 19% L1, 9% L4, 3% L5 and 1% T12, the remainder arising directly from the femoral nerve; the L2-L3 region is situated 4.6 cm on average above the iliac crest. Discussion: The region facing the L2-L3 space enables access to more than 50% of the psoas nerve branches. Injection via a posterior approach situated in adults 4.6 cm above the iliac crest and identified fluoroscopically is the most reliable access. This will avoid injury to the ureter which lies lower. Conclusion: This anatomy study described a new more effective less dangerous approach for botulinum toxin injections into the psoas muscle

Purpose: Isolated paralysis of the great toe long extensor is a rare complication of leg fractures. In certain patients, an erroneous diagnosis of compartment syndrome or muscle incarceration may be made. Material and methods: We dissected ten fresh cadavers. Results: The great toe long extensor was innervated by a branch of the deep fibular nerve which arose 15 cm from the talocrural joint space and directly in contact with the periosteum of the tibial shaft. In this localisation, the branch can be directly sectioned during trauma, reduction, or reaming. Discussion: We had one patient aged 30 years with a fracture of the mid third of the leg who was treated by centromedullar nailing. Postoperatively, this patient developed isolated paralysis of the great toe long extensor. The isolated neurogenic origin of this paralysis was confirmed by electromyography. Conclusion: This case is illustrative of direct injury of the great toe long extensor innervation, a cause which to our knowledge has not been previously described

The distal interphalangeal (DIP) joints of the fingers are prone to functional impotence in some degenerative diseases. In this case, different surgical techniques can be used, from DIP arthrodesis to joint denervation, much more confidential, which aims to preserve an already reduced mobility.

The four fingers (except the thumb) of 6 fresh hands from different cadavers were dissected under optic magnification. Two DIP joints were harvested from fresh dissected hands, in order to follow with the microscope the course of the nerve branchlets up to their articular entry. These two specimens were decalcified, and then embedded in paraffin. The blocks were serially cut in 5μm slices (1 slice each 250μm), which were observed at 25 and x100 magnification, after Masson’s trichrom staining.

A constant proximal articular branch, arising from the proper digital palmar nerve, was exclusively devoted to joint supply. This branch was located medially and arose in average at 7 mm from its entry point in the joint, where it was accompanied by small arterial branches. Before its entry into the inferomedial part of the DIP joint, it ran under the flexor digitorum profun-dus tendon. It then could divide into 2 or 3 branchlets. The proper digital palmar nerve abandoned, along its course, some nerve fibers to the tendinous synovium and neighboring structures. Then, ending its course, it gave off a distal articular branch, hidden among numerous cutaneous branches for the fingertip. The DIP joint nerve supply seems so under the exclusive dependence of the proper digital palmar nerve without any input from the dorsal side. On the histological slices, the nerves were mainly observed in peri- and intracapsular situation.

Could cutting these two articular nerves be sufficient to relieve pain from the DIP? This is what we are investigating through a clinical series; the first results are presented here.

Anatomically, bone consists of building blocks called osteons, which in turn comprise a central canal that contains nerves and blood vessels. This indicates that bone is a highly innervated and vascularized tissue. The function of vascularization in bone (development) is well-established: providing oxygen and nutrients that are necessary for the formation, maintenance, and healing. As a result, in the field of bone tissue engineering many research efforts take vascularization into account, focusing on engineering vascularized bone. In contrast, while bone anatomy indicates that the role of innervation in bone is equally important, the role of innervation in bone tissue engineering has often been disregarded. For many years, the role of innervation in bone was mostly clear in physiology, where innervation of a skeleton is responsible for sensing pain and other sensory stimuli. Unraveling its role on a cellular level is far more complex, yet more recent research efforts have unveiled that innervation has an influence on osteoblast and osteoclast activity. Such innervation activities have an important role in the regulation of bone homeostasis, stimulating bone formation and inhibiting resorption. Furthermore, due to their anatomical proximity, skeletal nerves and blood vessels interact and influence each other, which is also demonstrated by pathways cross-over and joint responses to stimuli. Besides those closely connected sytems, the immune system plays also a pivotal role in bone regeneration. Certain cytokines are important to attract osteogenic cells and (partially) inhibit bone resorption. Several leukocytes also play a role in the bone regeneration process. Overall, bone interacts with several systems. Aberrations in those systems affect the bone and are important to understand in the context of bone regeneration. This crosstalk has become more evident and is taken more into consideration. This leads to more complex tissue regeneration, but may recapitulate better physiological situations

Osteoarthritis (OA) is a highly prevalent degenerative joint disorder characterized by joint pain and physical disability. Aberrant subchondral bone induces pathological changes and is a major source of pain in OA. In the subchondral bone, which is highly innervated, nerves have dual roles in pain sensation and bone homeostasis regulation. The interaction between peripheral nerves and target cells in the subchondral bone, and the interplay between the sensory and sympathetic nervous systems, allow peripheral nerves to regulate subchondral bone homeostasis. Alterations in peripheral innervation and local transmitters are closely related to changes in nociception and subchondral bone homeostasis, and affect the progression of OA. Recent literature has substantially expanded our understanding of the physiological and pathological distribution and function of specific subtypes of neurones in bone. This review summarizes the types and distribution of nerves detected in the tibial subchondral bone, their cellular and molecular interactions with bone cells that regulate subchondral bone homeostasis, and their role in OA pain. A comprehensive understanding and further investigation of the functions of peripheral innervation in the subchondral bone will help to develop novel therapeutic approaches to effectively prevent OA, and alleviate OA pain. Cite this article: Bone Joint Res 2022;11(7):439–452

Introduction. The objective of the work is construction of a multi-bioactive scaffold based on that allows a space/time control over the regeneration of damaged bones by Medication-Related Osteonecrosis of the Jaw using a minimal invasive approach based on the injection of the fast-degrading pro neuro and angiogenic ELR (Elastin-Like Recombinamers) based hydrogels. Method. Chemical crosslinking facilitated the creation of multi-bioactive scaffolds using ELRs with reactive groups. Cell-loaded multi-bioactive scaffolds, prepared and incubated, underwent evaluation for adhesion, proliferation, angiogenic, and neurogenic potential. In vitro assessments utilized immunofluorescence staining and ELISA assays, while live-recorded monitoring and live-dead analysis ensured cytocompatibility. In rat and rabbit models, preformed scaffolds were subcutaneously implanted, and the regenerative process was evaluated over time. Rabbit models with MRONJ underwent traditional or percutaneous implantation, with histological evaluation following established bone histological techniques. Result. A 3D scaffold using ELR that combines various peptides with different degradation rates to guide both angiogenesis and neurogenesis has been developed. Notably, scaffolds with different degradation rates promoted distinct patterns of vascularization and innervation, facilitating integration with host tissue. This work demonstrates the potential for tailored tissue engineering, where the scaffold's bioactivities and degradation rates can control angiogenesis and neurogenesis. In an animal model of medication-related osteonecrosis of the jaw (MRONJ), the scaffold showed promising results in promoting bone regeneration in a necrotic environment, as confirmed by histological and imaging analyses. This study opens avenues for novel tissue-engineering strategies where precise control over vascularization and nerve growth is crucial. Conclusion. A groundbreaking dual approach, simultaneously targeting angiogenesis and innervation, addresses the necrotic bone in MRONJ syndrome. Vascularization and nerve formation play pivotal roles in driving reparative elements for bone regeneration. The scaffold achieves effective time/space control over necrotic bone regeneration. The authors are grateful for funding from the Spanish Government (PID2020-118669RA-I00)

Osteoarthritis (OA) is a common cause of chronic pain. Subchondral bone is highly innervated, and bone structural changes directly correlate with pain in OA. Mechanisms underlying skeletal–neural interactions are under-investigated. Bone derived axon guidance molecules are known to regulate bone remodelling. Such signals in the nervous system regulate neural plasticity, branching and neural inflammation. Perturbation of these signals during OA disease progression may disrupt sensory afferents activity, affecting tissue integrity, nociception, and proprioception. Osteocyte mechanical loading and IL-6 stimulation alters axon guidance signalling influencing innervation, proprioception, and nociception. Human Y201 MSC cells, embedded in 3D type I collagen gels (0.05 × 106 cell/gel) in 48 well plastic or silicone (load) plates, were differentiated to osteocytes for 7 days before stimulation with IL-6 (5ng/ml) with soluble IL-6 receptor (sIL-6r (40ng/ml) or unstimulated (n=5/group), or mechanically loaded (5000 μstrain, 10Hz, 3000 cycles) or not loaded (n=5/group). RNA extracted 1hr and 24hrs post load was quantified by RNAseq whole transcriptome analysis (NovaSeq S1 flow cell 2 × 100bp PE reads and differentially expressed neurotransmitters identified (>2-fold change in DEseq2 analysis on normalised count data with FDR p<0.05). After 24 hours, extracted IL-6 stimulated RNA was quantified by RT-qPCR for neurotrophic factors using 2–∆∆Ct method (efficiency=94-106%) normalised to reference gene GAPDH (stability = 1.12 REfinder). Normally distributed data with homogenous variances was analysed by two-tailed t test. All detected axonal guidance genes were regulated by mechanical load. Axonal guidance genes were both down-regulated (Netrin1 0.16-fold, p=0.001; Sema3A 0.4-fold, p<0.001; SEMA3C (0.4-fold, p<0.001), and up-regulated (SLIT2 2.3-fold, p<0.001; CXCL12 5-fold, p<0.001; SEMA3B 13-fold, p<0.001; SEMA4F 2-fold, p<0.001) by mechanical load. IL6 and IL6sR stimulation upregulated SEMA3A (7-fold, p=0.01), its receptor Plexin1 (3-fold, p=0.03). Neutrophins analysed in IL6 stimulated RNA did not show regulation. Here we show osteocytes regulate multiple factors which may influence innervation, nociception, and proprioception upon inflammatory or mechanical insult. Future studies will establish how these factors may combine and affect nerve activity during OA disease progression

Abstract. OBJECTIVE. Changes in subchondral bone are one of few disease characteristics to correlate with pain in OA. 1. Profound neuroplasticity and nociceptor sprouting is displayed within osteoarthritic (OA) subchondral bone and is associated with pain and pathology. 2. The cause of these neural changes remains unestablished. Correct innervation patterns are indispensable for bone growth, homeostasis, and repair. Axon guidance signalling factor, Sema3A is essential for the correct innervation patterning of bony tissues. 3. , expressed in osteocytes. 4. and known to be downregulated in bone OA mechanical loading. 5. Bioinformatic analysis has also shown Sema3a as a differentially expressed pathway by bone in human OA patients. 6. HYPOTHESIS: Pathological mechanical load and inflammation of bone causes dysregulation of Sema3A signalling leading to perturbed sensory nerve plasticity and pain. METHODS. Human KOLF2-C1 iPSC derived nociceptors were generated by TALEN-mediated insertion of transcription factors NGN2+Brn3A and modified chambers differentiation protocol to produce nociceptor-like cells. Nociceptor phenotype was confirmed by immunocytochemistry. Human Y201-MSC cells were embedded in 3D type-I collagen gels (0.05 × 106 cell/gel), in 48-well plates and silicone plates, were differentiated to osteocytes for 7 days before stimulation with IL-6 (5ng/ml) and soluble IL-6 receptor (sIL-6r (40ng/ml), IL6/sIL6r and mechanical load mimetic Yoda1 (5μM) or unstimulated (n=5/group) (48-well plates) or were mechanically loaded in silicone plates (5000μstrain, 10Hz, 3000 cycles) or not loaded (n=5/group). Conditioned media transfer was performed from osteocyte to nociceptor cultures assessed by continuous 24-hour phase contrast confocal microscopy. 24-hours after stimulation RNA was quantified by RT-qPCR (IL6) or RNAseq whole transcriptome analysis/DEseq2 analysis (Load). Protein release was quantified by ELISA. Normally distributed data with homogenous variances was analysed by two-tailed t test. RESULTS. IPSC-derived nociceptor-like cells display elongated (>5mm) dendritic projections and nociceptive molecular markers such as TUJ1, PrPH and Neun and TrkA. Sema3A signalling ligands were expressed in 100% of osteocyte cultures. Mechanical loading regulated the Sema3 pathway; Sema3A (0.4-fold, p<0.001), Sema3B (13-fold, p<0.001), Sema3C (0.4-fold, p<0.001). Under inflammatory stimulation by IL6/IL6sR, SEMA3A (7-fold, p=0.01) and receptor Plexin1 (3-fold, p=0.03) show significant regulation. Sema3A protein release showed a significant downregulation of Sema3A release by IL6/sIL6r+Yoda1 (2-fold, p=0.02). Continuous 24-hour phase contrast confocal microscopy measuring the number of extending/retreating dendritic projections revealed that sensory nerve cultures exposed to media from osteocytes stimulated with IL-6/sIL-6R+Yoda1 displayed significantly more invading dendritic projections (p=0.0175, 12-fold±SEM 3.5) across 3 random fields of view within a single stimulated neural culture and significantly fewer retracting dendritic projections (p=0.0075, 2-fold±SEM 0.33) compared to controls. CONCLUSIONS. Here we show osteocytic regulation of Sema3A under pathological mechanical loading and the ability of media pathologically loaded osteocyte cultures to induce the branching and invasion of cultured nociceptor-like cells as displayed in OA subchondral bone. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Tendinopathy is the most common form of chronic tendon disorders, accounting for up 30% of all musculoskeletal clinic visits [1]. In tendon disease, the largely avascular tendon tissue often becomes hypervascularized and fibrotic [2]. As blood vessel growth and angiogenic signaling molecules are often induced by the lack of adequate nutrients and oxygen, hypoxic signaling is speculated to be a root cause of tendon neovascularization and tendinopathy [3,4,5]. However, how the vascular switch is initiated in tendons, and how vascularization contributes to tendon pathology remains unknown. In this talk, we provide evidence that HIF-1α is implicated in tendon disease and HIF-1α stabilization in human tendon cells induces vascular recruitment of endothelial cells via VEGFa secretion. More interesting, HIF-1α stabilization in tendon cells in vivo, seems to recapitulate all main features of fibrotic human tendon disease, including vascular ingrowth, matrix disorganization, changes in tissue mechanics, cell proliferation and innervation. Surprisingly, in vivo knock-out of VEGFa rescued angiogenesis in the tendon core but it did not affect tendon mechanical properties and tissue pathophysiological changes, suggesting that blood vessels ingrowth might not be a primary cause but a consequence of HIF-1α activation

The suprascapular nerve is an ideal target for nerve blockade to alleviate shoulder pain given its widespread innervation to the shoulder girdle. Many techniques have been described. To widen the availability of this treatment we investigate whether an anatomical landmark technique can be easily learned by novice injectors to provide efficacious blockade. Five injectors were recruited with varying experience; from the novice medical student to an orthopaedic consultant. Five torsos (10 shoulders) were used. A single page of written instruction and illustration of the Dangoisse landmark technique was provided prior to injection of a Thiel embalmed cadaver bilaterally. A pre-mixed injectate with blue dye was used. Cadavers were dissected and the presence or absence of dye staining reported by 3 observers and a consensus agreement reached. Dissection demonstrated diffuse staining in the suprascapular fossa. 90% of shoulders were found to have adequate staining of the suprascapular nerve directly, or its distal branches, in a manner which would provide adequate anaesthesia. The inter-observer agreement was good (k = 0.73) for staining at the supraspinous fossa and excellent (k=0.87) for staining distally. The technique was easily performed by novice injectors with a 100% success rate. We demonstrate that this technique is reproducible by a range of clinicians to effectively provide anaesthesia of the SScN. The main risks are ineffective block (10% in this series) and of intravascular injection. Within a resource strained healthcare environment greater uptake of this technique is likely to be of benefit to a wider array of patients

Purpose of the study: There is no satisfactory surgical solution for symptomatic osteoarthritis of the elbow joint with preserved functional motion if arthroplasty is not indicated (age, functional demand). The same is true for resistant epicondylalgia. The joint denervation techniques applied for the wrist and proximal inter-phalangeal joints have demonstrated their efficacy. We conducted an anatomic study of elbow innervation as a preliminary step to the development of a standardized surgical procedure for complete denervation of the elbow compartment. Material and methods: The study was conducted on 15 right and left unprepared fresh cadaver specimens. A standardized dissection method was used. The terminal branches of the brachial plexus were dissected proximally to distally under magnification, from the root of the arm to the mid third of the forearm. Results: Innervation of the medial compartment arose: anteriorly, from one of the two capsuloperiosteal branches arising from the medial nerve; in the epitrochleo-olecraneal gutter, from capsular branches issuing from the trunk of the radial nerve at the root of the arm and running with the ulnar nerve. The innervation of the lateral compartment arose: anteriorly, from an inconstant capsular branch issuing from the musculo-cutaneous nerve arising 4 to 7 cm downstream from the joint space and running between the bones. In the other cases, this zone was innervated by a nerve branch coming from the dorsal cutaneous nerve of the forearm issuing from the radial nerve. This branch innervated the apex of the laeral epicondyle in all cases. The posterior part of the lateral compartment was constantly innervated by a branch arising from the radial nerve in the proximal part of the arm, running between the deep hed of the triceps and the vastus lateralis, giving rise of nerves innervating the joint and terminating in the body of the anconeus muscle. Discussion: Our study enabled the description of new sources of elbow innervation not reported by Wilhelm. Conclusion: This systematization study of elbow joint innervation is a preliminary step to the development of a complete procedure for unicompartmental lateral or medial denervation of the elbow joint. The fields of application are the treatment of symptomatic osteoarthritis of the elbow joint in patients with preserved joint motion and resistant epicondylalgia

Introduction and Objective. Intervertebral disc (IVD) degeneration is one of the major contributors to low back pain, the leading cause of disability worldwide. This multifactorial pathological process involves the degradation of the extracellular matrix, inflammation, and cell loss due to apoptosis and senescence. While the deterioration of the extracellular matrix and cell loss lead to structural collapse of the IVD, increased levels of inflammation result in innervation and the development of pain. Amongst the known regulators of inflammation, toll-like receptors (TLRs) and more specifically TLR-2 have been shown to be specifically relevant in IVD degeneration. As strong post-transcriptional regulators, microRNAs (miRNAs) and their dysregulation has been connected to multiple pathologies, including degenerative diseases such as osteoarthritis and IVD degeneration. However, the role of miRNAs in TLR signalling in the IVD is still poorly understood and was hence investigated in this study. Materials and Methods. Human Nucleus pulposus (hNP) and Annulus fibrosus (hAF) cells (n=5) were treated with the TLR-2/6 specific agonist PAM2CSK4 (100 ng/mL for 6 hours) in order to activate the TLR2 signalling pathway. After the activation both miRNA and mRNA were isolated, followed by next-generation sequencing and qPCR analysis of proinflammatory cytokines respectively. Furthermore, cell supernatants were used to analyze the secretion of proinflammatory cytokines with enzyme-linked immunosorbent assay. TLR-2 knockdown (siRNA) cells were used as a control. Statistical analysis was conducted by performing Kolmogorov-Smirnov test and a two-tailed Student's t-test using GraphPad Prism version 9.0.2 for Windows (GraphPad Software, La Jolla California USA). Results. TLR-2 activation resulted in the induction of an inflammatory cell response, with a significant increase in gene expression of interleukin (IL)-6 (525 ± 180 fold change, p < 0.05) and IL-8 (7513 ± 1907 fold change, p < 0.05) and protein secretion of IL-6 (30.5 ± 8.1 pg/mL) and IL-8 (28.9 ± 5.4 pg/mL). TLR-2 activation was furthermore associated with changes in the miRNA profile of hNP and hAF cells. Specifically, we identified 10 differentially expressed miRNAs in response to TLR-2 activation, amongst which were miR-335–3p (1.45 log2 FC, p < 0.05), miR-125b-1–3p (0.55 log2 FC, p < 0.05), and miR-181a-3p (−1.05 log2 FC, p < 0.05). Conclusions. The identified miRNAs are known to be associated with osteoarthritis (miR-335-3p), inflammation and IVD degeneration (mir-125-1-3p and miR-181a-3p), but the link to TLR signalling has not been previously reported. Experiments to validate the identified miRNAs and elucidate their functional role are undergoing. The identification of these miRNAs provides an opportunity to further investigate miRNAs in the context of TLR activation and inflammation and to enhance our understanding of underlying molecular mechanisms behind disc degeneration, inflammation, and TLR dysregulation

Introduction: Increased cell senescence has been reported in the human intervertebral disc (IVD) and was associated with degenerative pathology, particularly herniation. Increased IVD innervation and blood vessel ingrowth is associated with disc degeneration and the development of back pain. This preliminary study examines whether there is a relationship between the prevalence of senescent IVD cells and the extent to which the tissue is innervated and/or vascularised. Methods: Specimens of herniated IVD (n=16 patients: aged 36–71) were stained for senescence associated β-galactosidase activity (SA β-gal), then snap frozen and cryosectioned prior to immunolocalisation procedures to detect nerves (NF200) or blood vessels (CD34). Stained sections were counterstained with DAPI to reveal cell nuclei. The proportion of SA β-gal +ve cells was scored and the extent of neural and blood vessel ingrowth semi-quantitated. Results: The proportion of SA β gal +ve IVD cells ranged from 6% – 91% (median=16%) and was significantly correlated with age. The degree of neural or blood vessel ingrowth ranged from tissue which contained numerous (i.e. ≥10) positive cells/cell processes to tissue which was completely aneural or avascular. However, there was no clear relationship between the presence of SA β-gal +ve IVD cells and IVD innervation or vascularisation. Conclusions: Cell senescence has been associated with up-regulated expression of catabolic enzymes, e.g. MMPs and increased synthesis of trophic cytokines, e.g. VEGF. Such cellular activity might by thought to contribute to the pathological ingrowth of nerves or blood vessels into the IVD. The data presented here, however, does not support such a hypothesis. Conflicts of Interest: None. Source of Funding: Institute of Orthopaedics, RJAH Orthopaedic Hospital

Background: Increased nerve growth into degenerated intervertebral discs is associated with discogenic low back pain [. 1. ]. Many of these growing nerves are in neo-vascularised areas of the tissue [. 1. ,. 2. ] and endothelial cells that penetrate the disc express neurotrophic factors [. 3. ]. Thus, disc neovascularisation and disc innervation may be closely linked. Whilst disc aggrecan has been found to inhibit sensory nerve growth in vitro [. 4. ], the effects of disc aggrecan on endothelial cells are unknown. Methods/Results: Adapting in vitro assays used previously [. 4. ], with HMEC-1 and EAhy-926 cell lines as models of endothelial cell growth, we found that disc aggrecan inhibited endothelial cell migration in a dose-dependent manner. Endothelial cells traversed over collagen substrates until they encountered disc aggrecan substrates (1mg/ml human aggrecan), where they either stopped migrating or, more commonly, changed their direction of movement and aligned to the collagen:aggrecan border (Figure 1). After reaching the aggrecan border, some endothelial cells also migrated away from the disc aggrecan. At lower concentrations of disc aggrecan (0.01mg/ml), no such inhibition of endothelial cell growth was seen. Conclusions: Loss of aggrecan, increased innervation and neovascularisation are all marked features of disc degeneration [. 1. ,. 2. ,. 5. ]. This study provides evidence that disc aggrecan inhibits endothelial migration and therefore supports a hypothesis that a loss of aggrecan from degenerated discs predisposes the tissue to vascular invasion

To assess the potential for IDET to ablate nerve fibres in an experimentally induced peripheral annular lesion. Intradiscal electrothermal therapy (IDET) is being increasingly used as a minimally-invasive treatment for discogenic low back pain, with success reported in up to 70% of cases. One proposed mechanism of IDET is ablation reported in up to 70% of cases. One proposed mechanism of IDET is ablation of nerve fibres in the peripheral annulus. An ovine model was used to assess the innervation of peripheral annular lesions and the potential for IDET to denervate this region of the disc. Postero-lateral annular incisions were made in 32 lumbar discs of 16 sheep. At twelve weeks the sheep underwent IDET at one level and a sham treatment at the other level. IDET was performed using a modified Intradiscal Catheter (SpineCath, Oratec Interventions Inc., Menlo Park, CA). The spines were harvested at intervals up to six months. Histological sections of the discs were stained with H& E and an antibody to the general neuronal marker PGP 9.5. Vascular granulation tissue consistent with a healing posterior annular tear was observed in all incised discs from 12 weeks, extending to an average depth of 850 μm at 0 weeks to 690 μm at 6 months. PGP 9.5 positive nerve fibres were clearly identified outside the discs but were scarce within the discs. Nerves were identified up to 300 μm inside the annulus, from the earliest time point, and there was a trend towards less innervation with time. There were no fewer nerve fibres identified in those specimens that had undergone IDET. Specimens obtained six weeks after IDET showed evidence of thermal necrosis in the inner annulus, sparing the periphery of the disc. The reported benefit from IDET appears to be related to factors other than denervation. Thermal necrosis within the annulus six weeks after IDET