Regulatory Mechanism between Ferritin and Mitochondrial Reactive Oxygen Species in Spinal Ligament-Derived Cells from Ossification of Posterior Longitudinal Ligament Patient.

Primary spinal ligament-derived cells (SLDCs) from cervical herniated nucleus pulposus tissue (control, Ctrl) and ossification of the posterior longitudinal ligament (OPLL) tissue of surgical patients were analyzed for pathogenesis elucidation. Here, we found that decreased levels of ferritin and increased levels of alkaline phosphatase (ALP), a bone formation marker, provoked osteogenesis in SLDCs in OPLL. SLDCs from the Ctrl and OPLL groups satisfied the definition of mesenchymal stem/stromal cells. RNA sequencing revealed that oxidative phosphorylation and the citric acid cycle pathway were upregulated in the OPLL group. SLDCs in the OPLL group showed increased mitochondrial mass, increased mitochondrial reactive oxygen species (ROS) production, decreased levels of ROS scavengers including ferritin. ROS and ferritin levels were upregulated and downregulated in a time-dependent manner, and both types of molecules repressed ALP. Osteogenesis was mitigated by apoferritin addition. We propose that enhancing ferritin levels might alleviate osteogenesis in OPLL.

Exosomal miR-140-5p inhibits osteogenesis by targeting IGF1R and regulating the mTOR pathway in ossification of the posterior longitudinal ligament.

BACKGROUND: Ossification of the posterior longitudinal ligament (OPLL) is a disabling disease whose pathogenesis is still unclear, and there are no effective cures or prevention methods. Exosomal miRNA plays an important role in the osteogenesis of ectopic bone. Therefore, we focused on the downregulation of miR-140-5p in OPLL cell-derived exosomes to explore the mechanism by which exosomal miR-140-5p inhibits osteogenesis in OPLL. RESULTS: Exosomes were isolated by differential centrifugation and identified by transmission electron microscopy, nanoparticle tracking analysis, and exosomal markers. Exosomal RNA was extracted to perform miRNA sequencing and disclose the differentially expressed miRNAs, among which miR-140-5p was significantly downregulated. Confocal microscopy was used to trace the exosomal miR-140-5p delivered from OPLL cells to human mesenchymal stem cells (hMSCs). In vitro, we verified that exosomal miR-140-5p inhibited the osteoblast differentiation of hMSCs by targeting IGF1R and suppressing the phosphorylation of the IRS1/PI3K/Akt/mTOR pathway. In vivo, we verified that exosomal miR-140-5p inhibited ectopic bone formation in mice as assessed by micro-CT and immunohistochemistry. CONCLUSIONS: We found that exosomal miR-140-5p could inhibit the osteogenic differentiation of hMSCs by targeting IGF1R and regulating the mTOR pathway, prompting a further potential means of drug treatment and a possible target for molecular therapy of OPLL.

Enhanced magnetic resonance imaging manifestations of paediatric intervertebral disc calcification combined with ossification of the posterior longitudinal ligament: case report and literature review.

BACKGROUND: Since the first description of paediatric intervertebral disc calcification (IDC) by Báron in 1924, only approximately 400 cases have been reported in the literature. Paediatric IDC combined with ossification of the posterior longitudinal ligament (OPLL) is an even rarer condition, with only 8 cases described in detail to date. In this paper, we present a review of the disease characteristics described in the relevant English language literature and discuss the possible mechanisms of lesion enhancement in contrast-enhanced magnetic resonance imaging (MRI). CASE PRESENTATION: In May 2020, a 6-year-old Han nationality girl presented with the chief complaint of neck pain that had lasted for a week. She did not report a history of trauma or a past illness. On admission, there was no personal and family history, congenital diseases, or non-specific infections such as tuberculosis, among others. Further physical examination revealed that the movement of her cervical spine was limited. Computed tomography (CT) and MRI revealed ossification of the intervertebral discs and posterior longitudinal ligament (PLL) at the C4/5 levels and an absence of obvious spinal cord compression. When contrast-enhanced MRI was performed, significant enhancement was observed in the intervertebral discs and PLL at the C4/5 level. We adopted a non-interventional approach and performed an imaging re-examination 8 months later. Both the plain and contrast-enhanced MRI scans indicated swelling in the C4/5 intervertebral discs and disappearance of the previously observed enhancement in the nucleus pulposus (NP) and PLL at the corresponding levels; CT examination revealed that the ossified lesions had been completely resorbed. CONCLUSION: Obvious lesion enhancement in contrast-enhanced MRI is an extremely rare manifestation of paediatric IDC combined with OPLL. However, the exact mechanisms of this phenomenon remain unclear. We surmise that it may be caused by a series of biophysical changes related to vertebral endplate injury and repair, but further research will be required for in-depth investigation.

Effect of teriparatide on ligamentum flavum mesenchymal stem cells isolated from patients with ossification of the posterior longitudinal ligament.

Ossification of the posterior longitudinal ligament (OPLL) within the spinal canal sometimes leads to severe myelopathy. Teriparatide (TPD) is a recombinant human parathyroid hormone (PTH) (1-34), which promotes osteogenesis of mesenchymal stem cells (MSCs) via PTH 1 receptor (PTH1R). Although ligamentum flavum (LF)-MSCs from patients with OPLL have a high osteogenic potency, the effect of TPD on them remains unknown. In this study, we determined PTH1R expression in LF-MSCs from patients with OPLL and investigated whether TPD promotes osteogenic differentiation in them. First, LF-MSCs were isolated from patients with OPLL and cervical spondylotic myelopathy (CSM) (controls). Cultured LF-MSCs were treated with different concentrations of TPD on days 0, 7, and 14. On day 21, osteogenic gene expression was quantified. Mineralization was measured based on optical density after Alizarin Red S staining. LF-MSCs from both groups expressed PTH1R at the same level. TPD did not enhance osteogenic gene expression and mineralization in LF-MSCs from both groups. TPD did not promote the osteogenic differentiation of LF-MSCs from patients with OPLL. Thus, it may be safe for patients with OPLL. However, further confirmation of our results with in vivo studies is necessary.

Ossification of the cervical anterior longitudinal ligament is an underdiagnosed cause of difficult airway: a case report and review of the literature.

BACKGROUND: Ossification of the anterior longitudinal ligament (OALL) of the cervical spine is a common, but rarely symptomatic, condition mostly observed in the geriatric population. Although the condition usually requires no intervention, it could lead to a difficult airway and compromise the patient's safety. CASE PRESENTATION: Here, we describe the case of a 50-year-old man with cervical myelopathy and OALL that resulted in difficult endotracheal intubation after induction of anesthesia. Radiography and magnetic resonance imaging findings showed OALL, with prominent osteophytes involving four cervical vertebrae, a bulge in the posterior pharyngeal wall, and a narrow pharyngeal space. Airtraq® laryngoscope-assisted intubation was accomplished with rapid induction under sevoflurane-inhaled anesthesia. CONCLUSION: Anesthesiologists should understand that OALL of the cervical spine could cause a difficult airway. However, it is difficult to recognize asymptomatic OALL on the basis of routine airway evaluation guidelines. For susceptible populations, a thorough evaluation of the airway, based on imaging studies and a history of compression symptoms, should be considered whenever possible. In case of unanticipated difficult intubation, anesthesiologists should refer to guidelines for unanticipated difficult airway management and identify OALL of the cervical spine as the cause.

On the importance of the innervation of the human cervical longitudinal ligaments at vertebral level.

PURPOSE: In our aging society, the prevalence of degenerative spinal diseases rose drastically within the last years. However, up till now, the origin of cervical pain is incompletely understood. While animal and small cadaver studies indicate that a complex system of sensory and nociceptive nerve fibers in the anterior (ALL) and posterior longitudinal ligament (PLL) at the level of the intervertebral disc might be involved, there is a lack of data exploring whether such a network exists and is equally distributed within the cervical vertebrae (VB). We, therefore, aimed to investigate the spatial distribution of the mentioned nerve networks in human tissue. METHODS: We performed macroscopic (Sihler staining, Spalteholz technique, and Plastination) and microscopic (immunohistochemistry for PGP 9.5 and CGRP) studies to characterize spatial differences in sensory and nociceptive innervation patterns. Therefore, 23 human body donors were dissected from level C3-C6. RESULTS: We could show that there is a focal increase in sensory and nociceptive nerve fibers at the level of C4 and C5 for both ALL and PLL, while we observed less nerve fiber density at the level of C3 and C6. An anatomical vicinity between nerve and vessels was observed. CONCLUSION: To our knowledge, these findings for the first time report spatial differences in sensory and nociceptive nerve fibers in the human cervical spine at VB level. The interconnection between nerves and vessels supports the importance of the perivascular plexus. These findings might be of special interest for clinical practice as many patients suffer from pain after cervical spine surgery.

Endoplasmic reticulum stress regulates mechanical stress-induced ossification of posterior longitudinal ligament.

PURPOSE: The pathogenesis of ossification of posterior longitudinal ligament (OPLL) is not completely clear. Previous study has confirmed a single-pass type I endoplasmic reticulum (ER) membrane protein kinase (PERK), which is a major transducer of the ER stress, participates in the process of OPLL in vitro. This study aimed to demonstrate the role of ER stress in mechanical stress (MS)-induced OPLL. METHODS: The posterior longitudinal ligaments were collected intraoperatively. The expression of ER stress markers in ligament tissue samples was compared between OPLL and non-OPLL patients in vivo. Ligament fibroblasts were isolated and cultured. Loaded by MS, the expression of ER stress markers in fibroblasts deriving from non-ossified areas of the ligament tissues from OPLL patients was detected. The influence of inhibition of ER stress on MS-induced OPLL and activation of mitogen-activated protein kinase (MAPK) pathways by MS was also investigated. RESULTS: We confirmed the ER stress markers were highly expressed in non-ossified areas of the ligament tissues from OPLL patients but could barely be detected in the ligaments from non-OPLL patients in vivo. We also found ER stress could be activated by MS during the process of OPLL in vitro. Moreover, inhibition of ER stress could hinder MS-induced OPLL and activation of MAPK signaling pathways by MS in vitro. CONCLUSION: Activated ER stress was observed in OPLL patients both in vitro and in vivo. Mechanical stress could activate ER stress response in posterior longitudinal ligament fibroblasts and further promote OPLL in vitro. In this process, ER stress might work through the MAPK signaling pathways. These slides can be retrieved under Electronic Supplementary Material.

Combined use of leptin and mechanical stress has osteogenic effects on ossification of the posterior longitudinal ligament.

PURPOSE: To evaluate the effects of leptin/leptin receptor (LepR) combined with mechanical stress on the development of ossification of the posterior longitudinal ligament (OPLL), which is a disease characterized by ectopic bone formation of the posterior longitudinal ligament (PLL) and can lead to radiculopathy and myelopathy. METHODS: Six human samples of the PLL were analyzed for the expression of leptin and LepR by RT-PCR and western blotting. PLL cells were stimulated with leptin and mechanical stress delivered via a Flexcell tension system, and osteogenic differentiation was evaluated by RT-PCR and western blotting analysis of osteogenic marker expression as well as by alkaline phosphatase (ALP) staining and alizarin red S staining. Activation of mitogen-activated protein kinase (MAPK), Janus kinase (JAK) 2-signal transducer, activator of transcription (STAT) 3 and phosphatidylinositol 3-kinase (PI3K)-Akt was evaluated by western blotting. RESULTS: Samples from the OPLL group had higher LepR mRNA and protein levels and lower leptin levels than those from healthy controls. Exposure to leptin and Flexcell increased the number of ALP-positive cells and calcium nodules in a dose-dependent manner; this effect was accompanied by upregulation of the osteogenic markers osteocalcin, runt-related transcription factor 2 (RUNX2) and osteopontin. Extracellular signal-regulated kinase, P38 MAPK, JAK2, STAT3, PI3K and Akt signaling, was also activated by the combined effects of leptin and mechanical stress. CONCLUSIONS: Leptin and LepR are differentially expressed in OPLL tissues, and the combined use of leptin/LepR and mechanical stress promotes osteogenic differentiation of PLL cells via MAPK, JAK2-STAT3 and PI3K/Akt signaling. These slides can be retrieved under Electronic Supplementary Material.

Myelopathy associated with instability consequent to resection of ossification of anterior longitudinal ligament in DISH.

PURPOSE: The presence of prominent OALL (ossification of anterior longitudinal ligament) in the anterior cervical spine has been implicated as a cause of dysphagia. Surgical resection of the OALL is considered effective for the management of diffuse idiopathic skeletal hyperostosis (DISH)-related dysphagia. Although many reports have been published on DISH-related dysphagia, no cases of postoperative cervical instability have been reported thus far. We present a case in which the patient developed myelopathy associated with instability consequent to resection of OALL in DISH. METHODS: A 62-year-old man presented with progressive dysphagia that persisted for a year. The patient's symptoms were successfully resolved by resection of OALL. Five years after the surgery, the dysphagia resurfaced and was found to be caused by the regrowth of the OALL. A repeat surgery was performed, and the dysphagia disappeared. Eleven months after the second surgery, he visited the hospital with progressive quadriparesis and pain in the cervical region. RESULTS: Nine-month follow-up radiologic study revealed cervical instability at the level of C5-6 resulting in myelopathy. The patient underwent decompressive laminectomy and posterior fusion surgery. CONCLUSION: Surgical resection of DISH-related dysphagia typically yields excellent outcomes, but our experience in this case highlights the possibility of OALL regrowth and subsequent cervical instability after resection of OALL.

Effects of multilevel posterior ligament dissection after spinal instrumentation on adjacent segment biomechanics as a potential risk factor for proximal junctional kyphosis: a biomechanical study.

BACKGROUND: Spinous processes and posterior ligaments, such as inter- and supraspinous ligaments are often sacrificed either deliberately to harvest osseous material for final spondylodesis e.g. in deformity corrective surgery or accidentally after posterior spinal instrumentation. This biomechanical study evaluates the potential destabilizing effect of a progressive dissection of the posterior ligaments (PL) after instrumented spinal fusion as a potential risk factor for proximal junctional kyphosis (PJK). METHODS: Twelve calf lumbar spines were instrumented from L3 to L6 (L3 = upper instrumented vertebra, UIV) and randomly assigned to one of the two study groups (dissection vs. control group). The specimens in the dissection group underwent progressive PL dissection, followed by cyclic flexion motion (250 cycles, moment: + 2.5 to + 20.0 Nm) to simulate physical activity and range of motion (ROM) testing of each segment with pure moments of ±15.0 Nm after each dissection step. The segmental ROM in flexion and extension was measured. The control group underwent the same loading and ROM testing protocol, but without PL dissection. RESULTS: In the treatment group, the normalized mean ROM at L2-L3 (direct adjacent segment of interest, UIV/UIV + 1, PJK-level) increased to 104.7%, 107.3%, and 119.4% after dissection of the PL L4-L6, L3-L6, and L2-L6, respectively. In the control group the mean ROM increased only to 103.2%, 106.7%, and 108.7%. The ROM difference at L2-L3 with regard to the last dissection of the PL was statistically significant (P = 0.017) and a PL dissection in the instrumented segments showed a positive trend towards an increased ROM at UIV/UIV + 1. CONCLUSIONS: A dissection of the PL at UIV/UIV + 1 leads to a significant increase in ROM at this level which can be considered to be a risk factor for PJK and should be definitely avoided during surgery. However, a dissection of the posterior ligaments within the instrumented segments while preserving the ligaments at UIV/UIV + 1 leads to a slight but not significant increase in ROM in the adjacent cranial segment UIV/UIV + 1 in the used experimental setup. Using this experimental setup we could not confirm our initial hypothesis that the posterior ligaments within a long posterior instrumentation should be preserved.

Natural history of the ossification of cervical posterior longitudinal ligament: a three dimensional analysis.

PURPOSE: Three-dimensional (3D) imaging using computed tomography (CT) has made it possible to accurately evaluate ossification of the posterior longitudinal ligament (OPLL). Recently, we developed a novel technique to measure ossification volume using the 3D analysis. The purpose of this study was to investigate the natural course of OPLL and the risk factors for volume progression. METHODS: Forty-one patients (22 males and 19 females) diagnosed with cervical OPLL who had been non-surgically treated were included in this study. We evaluated clinical examination, radiological findings, and the volume of ossified lesions during at least 1-year intervals. Furthermore, we performed risk factor analysis for OPLL volume progression. RESULTS: The mean ossification volume was 2047.4 ± 1437.3 mm3 in the first examination and 2201.0 ± 1524.1 mm3 in the final examination, indicating a significant increase during the follow-up period (p < 0.001). The mean annual rate of lesion increase was 4.1 ± 2.7%. Univariate regression analysis demonstrated significant relationships between the annual rate of lesion increase and age (ß = -0.48; p = 0.001), body weight (BW) (ß = 0.36; p = 0.02), and body mass index (BMI) (ß = 0.35; p = 0.03). Furthermore, age was the only significant predictor of OPLL progression (R2 = 0.23; p = 0.001) in multivariate liner regression analysis. CONCLUSIONS: Younger age, higher BW, and higher BMI are predictors of OPLL progression. Younger age is the most significant predictor in non-surgically treated patients.

Ossification of the posterior ligament is mediated by osterix via inhibition of the ß-catenin signaling pathway.

Ossification of the posterior longitudinal ligament (OPLL) involves ectopic calcification of the spinal ligament preferentially at the cervical spine. OPLL is associated with different diseases and occurs by endochondral ossification, which is associated with the activity of different transcription factors. However, the pathogenesis of OPLL remains unclear. Here, we investigated the role of osterix (Osx), a transcription factor that functions downstream of Runx2 and is an important regulator of osteogenesis, in the process of OPLL in a dexamethasone (Dex)-induced model of spinal ligament ossification. Our results showed that Osx is upregulated in patients with OPLL and during the ossification of ligament cells in parallel with the upregulation of osteogenic markers including osteocalcin (OCN), alkaline phosphatase (ALP) and collagen-1 (Col-1). Dex-induced ossification of ligament cells was associated with the downregulation and inactivation of ß-catenin, and these effects were offset by Osx knockdown. Activation of ß-catenin signaling abolished the effect of Dex on ossification and the upregulation of osteogenic markers. Taken together, our results suggest that OPLL is mediated by Osx via a mechanism involving the Wnt/ß-catenin signaling pathway, providing a basis for further research to identify potential targets for the treatment of OPLL.

Clinical image of anterior longitudinal ligament syndrome - an interdisciplinary problem.

INTRODUCTION: Approximately 1.5-2.5% of all patients treated due to discopathy have anterior longitudinal ligament lesions. An intervertebral disc moving under the anterior longitudinal ligament causes ligament displacement and irritation of the autonomic nervous system structures, resulting in a disturbed function of the organs controlled by this system. Clinical image: Increased sympathetic system activity in the thoracic section may cause symptoms that mimic coronary heart disease while irritation of the autonomic structures in the lumbosacral section of the spine results in a clinical presentation similar to that of gastrointestinal or gynecological disorders. The clinical image of the most common disorders is presented. The diagnosis may be formulated with the use of magnetic resonance imaging. TREATMENT: pharmacotherapy is ineffective. Physiotherapy and rehabilitation constitute a method of choice in the treatment of this syndrome.

Connexin 43 promotes ossification of the posterior longitudinal ligament through activation of the ERK1/2 and p38 MAPK pathways.

Although cervical ossification of the posterior longitudinal ligament (OPLL) is one of the most common spinal diseases, the pathogenic mechanism is still not fully understood. Abnormal mechanical stress distribution is believed to be one of the main causes of OPLL. We have previously found that mechanical stress can up-regulate connexin 43 (Cx43) expression in ligament fibroblasts; this transduces mechanical signals to promote osteoblastic differentiation. In the present study, in order to explore further the intracellular mechanisms of Cx43-induced osteoblast differentiation of ligament fibroblasts, we investigate the potential roles of the osteogenic signaling pathway components ERK1/2, p38 MAPK and JNK in Cx43-mediated mechanical signal transduction. We first confirm higher Cx43 levels in both in vivo ligament tissue from OPLL patients and in vitro cultured OPLL cells. We find that ERK1/2, p38 MAPK and the JNK pathway are all activated both in vivo and in vitro. The activation of these signals was dependent upon Cx43, as its knock-down resulted in diminished mechanical effects and reduced signaling. Moreover, its knock-down almost reversed the osteogenic effect of mechanical stress on ligament fibroblasts and the blocking of the ERK1/2 and p38 MAPK pathways but not the JNK pathway, partly diminished this effect. Therefore, Cx43, which is up-regulated by mechanical stress, seems to function partly via the activation of ERK1/2 and p38 MAPK signals to promote the osteoblastic differentiation of ligament fibroblasts.

Morphological changes in the human cervical intervertebral disc post trauma: response to fracture-type and degeneration grade over time.

PURPOSE: In the first 24 h post-intervertebral disc (IVD) trauma, up to 75 % cell death has been reported. In addition, burst fractures cause post-traumatic disc degeneration by elevated pro-apoptotic and pro-inflammatory gene transcription. Moreover, some patients have pre-trauma degenerative disc disease. The aim of the study was to assess histological changes and cell-death over a time period of up to 1 year caused by mechanical and structural factors. METHODS: 116 anterior portions of IVDs of the cervical spine were studied histologically by light microscopy and ultrastructurally by transmission electron microscopy (TEM). The group was investigated with regard to three main parameters: fracture mechanism (compressive vs. tensile/shear loads), degeneration grade (low vs. high) and endplate fracture (with vs. without). Disc architecture (e.g. ruptures) was studied histologically. Cell morphology was examined ultrastructurally to quantify cell-death, healthy and balloon cells. According to ultrastructural observations, two time-groups (up to 6 days vs. later) were established. Statistical analyses were carried out within and between time-groups. RESULTS: Histological changes were obvious in the annulus fibrosus where ruptures with haematoma were replaced by granulation tissue. Significant differences in cell-death were seen in the first few days due to different loads. In contrast to the more degenerated segments, low degenerated ones revealed significantly less cell death with time post-trauma. Interestingly, no difference was found between groups after the sixth day. Cell-death (mean 44 % for all investigated groups) remained high after day 6 post-trauma. CONCLUSION: IVDs retrieved from low grade degenerated segments revealed a significant recovery, with less cell-death and a partially restored disc matrix, although cell-death remained high. Long-term clinical studies of stabilized segments arising from different fracture mechanisms are required.

[Reoperation for cervical myelopathy duo to progressing ossification of the posterior longitudinal ligaments in patients with expansive laminoplasty].

OBJECTIVE: To retrospectively analyze the clinical data of the patients with reoperation for cervical myelopathy due to progressing ossification of the posterior longitudinal ligaments, with previous open-door expansive laminoplasty, and to evaluate the outcomes. METHODS: From May 2006 to July 2012, a retrospective study was performed on a consecutive series of 17 patients with previous open-door expansive laminoplasty, who had received the reoperation for cervical myelopathy due to progressing ossification of the posterior longitudinal ligaments. The reoperation was performed based on the clinical manifestations and segments of responsibility. The anterior approaches were performed in 12 cases, and the posterior approaches in 5 cases. The correlation between the clinical factors and Japanese Orthopedic Association (JOA) scores or the JOA recovery rate was evaluated by Pearson or Spearman correlation test. The pre- and post-operative JOA scores were analyzed by repeated measures ANOVA and the JOA recovery rates were compared with paired t test. RESULTS: The mean follow-up was 137.5 months (range 60-348 months). There were no serious complications after surgical procedures. There was one case that had C5 palsy in the first operation and had recovery after one week. Another case had C5 palsy in the reoperation with posterior approach, which had recovery at the end of 6 months postoperation. Three cases had the cerebrospinal fluid leakage of the reoperation, with two cases in the anterior approaches and one case in the posterior approach. There was no significant correlation between the clinical variables and JOA scores or JOA recovery rates. The JOA scores of the patients in the first operation were improved from 9.4±4.1 to 12.8±2.8 (P<0.01), and the JOA recovery rate was 45.6%. The JOA scores of the reoperation were improved from 10.2±2.8 to 12.7±2.4 (P<0.05) at the end of 6 months and 14.3±1.9 (P<0.01) by the last follow-up. There were significant differences between the JOA recovery rates by the last follow-up (63.2%) and at the end of 6 months (39.3%) of the reoperation or 45.6% of the first operation (P<0.01). CONCLUSION: The reoperation for cervical myelopathy duo to progressing ossification of the posterior longitudinal ligaments can significantly promote the recovery of the spinal cord, based on the clinical manifestations combined with segments of responsibility of the imaging.

Ossification of the Posterior Longitudinal Ligament: Imaging Findings in the Era of Cross-Sectional Imaging.

Imaging appearance and classification systems of ossification of the posterior longitudinal ligament (OPLL) on computed tomography and magnetic resonance imaging will be reviewed. Computed tomography evaluation most accurately demonstrates OPLL length and thickness, whereas magnetic resonance imaging has the advantage of demonstrating abnormal signal in the cord. Neurologic symptoms are most common in the cervical spine and are related to the degree of spinal stenosis and presence of cord edema. Surgical treatment usually involves cases of cervical OPLL and includes anterior or posterior decompression.

[Difficult Airway Management of a Patient with the Ossification of Anterior Longitudinal Ligament].

We report a case of difficult airway management (DAM) with the ossification of anterior longitudinal ligament (OALL). A 66-year-old man complained of pharyngeal discomfort. He was diagnosed with OALL, and planned to have a surgery under general anesthesia. We expected DAM due to the limitation of cervical mobility and airway obstruction caused by OALL. We succeeded in awake intubation with video laryngoscope and tracheal tube introducer.

Osteogenic lineage commitment of mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament.

Ectopic bone formation is thought to be responsible for ossification of the posterior longitudinal ligament of the spine (OPLL). Mesenchymal stem cells (MSCs) were isolated from spinal ligaments and shown to play a key role in the process of ectopic ossification. The purpose of this study was to explore the capacity of these MSCs to undergo lineage commitment and to assess the gene expression changes between these committed and uncommitted MSCs between OPLL and non-OPLL patients. Spinal ligament-derived cells were obtained from OPLL patients or patients with cervical spondylotic myelopathy (non-ossified) for comparison (n=8 in each group). MSCs from the two patient cohorts were evaluated for changes in colony forming ability; osteogenic, adipogenic and chondrogenic differentiation potential; and changes in gene expression following induction with lineage-specific conditions. We show that the osteogenic differentiation potential was significantly higher in MSCs from OPLL patients than in those from non-OPLL patients. In addition, alkaline phosphatase activity and several osteogenic-related genes expressions (bone morphogenetic protein 2, runt-related transcription factor 2 and alkaline phosphatase) were significantly higher in the OPLL group than in the non-OPLL group. However, single cell cloning efficiency, adipogenic and chondrogenic differentiation, and the expression of adipogenic and chondrogenic-related genes were equivalent between MSCs harvested from OPLL and non-OPLL patient samples. These findings suggest an increase in the osteogenic differentiation potential of MSCs from OPLL patients and that this propensity toward the osteogenic lineage may be a causal factor in the ossification in these ligaments.