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

1 . Voluntary activity of any given muscle in the hand is not an absolute indication of the state of the nerve which usually supplies it. 2. Significant variations in the standard pattern of innervation have been found in 20 per cent of 226 cases studied. 3. The pattern of innervation described in standard text-books occurred in only 33 per cent. of cases. 4. A striking variation is the supply of every thenar muscle by the ulnar nerve. In other cases the first dorsal interosseous muscle may be supplied by the median nerve. 5. In order to arrive at an accurate diagnosis when anomalous innervation is suspected, nerve blocks at appropriate levels are required. 6. Great care must be taken during operations to avoid damage to connections between the ulnar and the median nerves, especially in patients with anomalous innervation of the hand muscles

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

1. Pain and pressure sensibility has been studied in the fibrous articular ligament and synovial membrane of the knee joint in normal human subjects and in patients subjected to arthrotomy under local anaesthesia. 2. The fibrous ligament was found to be a highly sensitive structure, containing many spots which give rise to sensations of pain or pressure when stimulated mechanically or chemically. Synovial membrane was found to be a relatively insensitive structure which only occasionally contains pain-sensitive spots. 3. Histological examination of articular capsule from the knee joints of normal and sympathectomised cats shows that articular ligaments have a rich nerve plexus and a variety of specialised and unspecialised nerve endings, most of which are somatic in origin. Synovial membrane contains a more delicate nerve network and also a variety of nerve endings, the majority of which are autonomic in origin. But a substantial number of somatic nerves enter the synovial membrane, some of which terminate in nerve loops, globular endings or simple unspecialised endings. 4. Histological examination of normal human articular capsule shows that its innervation closely resembles that of the cat, but no sympathectomised human material was examined. 5. The bearing of these findings on the symptomatology of joint disease is discussed briefly

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

Using a rat model the characteristics of the sensory neurones of the dorsal-root ganglia (DRG) innervating the hip were investigated by retrograde neurotransport and immunohistochemistry.

Fluoro-Gold solution (FG) was injected into the left hip of ten rats. Seven days later the DRG from both sides between T12 and L6 were harvested. The number of FG-labelled calcitonin gene-related peptide-immunoreactive or isolectin B4-binding neurones were counted.

The FG-labelled neurones were distributed throughout the left DRGs between T13 and L5, primarily at L2, L3, and L4. Few FG-labelled isolectin B4-binding neurones were present in the DRGs of either side between T13 and L5, but calcitonin gene-related peptide-immunoreactive neurones made up 30% of all FG-labelled neurones.

Our findings may explain the referral of pain from the hip to the thigh or lower leg corresponding to the L2, L3 and L4 levels. Since most neurones are calcitonin gene-related peptide-immunoreactive peptide-containing neurones, they may have a more significant role in the perception of pain in the hip as peptidergic DRG neurones.

Our understanding of the origin of hip pain in degenerative disorders of the hip, including primary osteoarthritis, avascular necrosis and femoroacetabular impingement (FAI), is limited. We undertook a histological investigation of the nociceptive innervation of the acetabular labrum, ligamentum teres and capsule of the hip, in order to prove pain- and proprioceptive-associated marker expression. These structures were isolated from 57 patients who had undergone elective hip surgery (44 labral samples, 33 ligamentum teres specimens, 34 capsular samples; in 19 patients all three structures were harvested). A total of 15 000 histological sections were prepared that were investigated immunohistochemically for the presence of protein S-100, 68 kDa neurofilament, neuropeptide Y, nociceptin and substance P. The tissues were evaluated in six representative areas.

Within the labrum, pain-associated free nerve ending expression was located predominantly at its base, decreasing in the periphery. In contrast, the distribution within the ligamentum teres showed a high local concentration in the centre. The hip capsule had an almost homogeneous marker expression in all investigated areas.

This study showed characteristic distribution profiles of nociceptive and pain-related nerve fibres, which may help in understanding the origin of hip pain.

Cite this article: Bone Joint J 2013;95-B:770–6.

We obtained intervertebral discs with cartilage endplates and underlying cancellous bone at operation from patients with degenerative disc disease and then used immunohistochemical techniques to localise the nerves and nerve endings in the specimens. We used antibodies for the ubiquitous neuronal protein gene product 9.5 (PGP 9.5). Immunoreactivity to neuropeptide Y was used to identify autonomic nerves and calcitonin gene-related peptide (CGRP) and substance P to identify sensory nerves. Blood vessels were identified by immunoreactivity with platelet-endothelial cell-adhesion molecule (CD31; PECAM).

In a control group with no known history of chronic back pain, nerve fibres immunoreactive to PGP 9.5 and neuropeptide Y were most closely related to blood vessels, with occasional substance P and CGRP immunoreactivity. In patients with severe back pain and markedly reduced disc height, proliferation of blood vessels and accompanying nerve fibres was observed in the endplate region and underlying vertebral bodies. Many of these nerves were immunoreactive to substance P or CGRP, and in addition, substance P- and CGRP-immunoreactive nociceptors were seen unrelated to blood vessels. Quantification by image analysis showed a marked increase in CGRP-containing sensory nerve fibres compared with normal control subjects.

We speculate that a chemotactic response to products of disc breakdown is responsible for the proliferation of vascularity and CGRP-containing sensory nerves found in the endplate region and vertebral body adjacent to degenerate discs. The neuropeptides substance P and CGRP have potent vasodilatory as well as pain-transmitting effects. The increase in sensory nerve endings suggests increase in blood flow, perhaps as an attempt to augment the nutrition of the degenerate disc. The increase in the density of sensory nerves, and the presence of endplate cartilage defects, strongly suggest that the endplates and vertebral bodies are sources of pain; this may explain the severe pain on movement experienced by some patients with degenerative disc disease.

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