Conditioned medium from human dental pulp stem cells treats spinal cord injury by inhibiting microglial pyroptosis.

Human dental pulp stem cell transplantation has been shown to be an effective therapeutic strategy for spinal cord injury. However, whether the human dental pulp stem cell secretome can contribute to functional recovery after spinal cord injury remains unclear. In the present study, we established a rat model of spinal cord injury based on impact injury from a dropped weight and then intraperitoneally injected the rats with conditioned medium from human dental pulp stem cells. We found that the conditioned medium effectively promoted the recovery of sensory and motor functions in rats with spinal cord injury, decreased expression of the microglial pyroptosis markers NLRP3, GSDMD, caspase-1, and interleukin-1ß, promoted axonal and myelin regeneration, and inhibited the formation of glial scars. In addition, in a lipopolysaccharide-induced BV2 microglia model, conditioned medium from human dental pulp stem cells protected cells from pyroptosis by inhibiting the NLRP3/caspase-1/interleukin-1ß pathway. These results indicate that conditioned medium from human dental pulp stem cells can reduce microglial pyroptosis by inhibiting the NLRP3/caspase-1/interleukin-1ß pathway, thereby promoting the recovery of neurological function after spinal cord injury. Therefore, conditioned medium from human dental pulp stem cells may become an alternative therapy for spinal cord injury.

Focused ultrasound-mediated blood-brain barrier opening combined with magnetic targeting cytomembrane based biomimetic microbubbles for glioblastoma therapy.

Glioblastoma is the most common type of brain tumor. Due to the presence of the blood-brain barrier, the effects of chemotherapy have been unsatisfactory. The combination of focused ultrasound and microbubbles to reversibly open the blood-brain barrier is now considered a key factor in improving treatment outcomes of glioblastoma. In this study, we developed bionic drug delivery microbubbles, which in combination with focused ultrasound had an obvious inhibitory effect on glioblastoma. We extracted the brain microvascular cell membranes, combined them with lipid components, and loaded them with superparamagnetic iron oxide and doxorubicin to prepare biomimetic drug delivery microbubbles (FeDOX@cellMBs). We demonstrated that FeDOX@cellMBs retained the intrinsic properties of loading, such as magnetic properties and drug toxicity, both in vitro and in vivo. FeDOX@cellMBs exhibited good tumor targeting and uptake under the combined action of magnetic and focused ultrasound. Importantly, the FeDOX@cellMBs demonstrated excellent internal stability and effectively inhibited tumor growth in orthotopic glioblastoma mice. Finally, organ H&E staining confirmed that FeDOX@cellMBs were safe for use. In conclusion, FeDOX@cellMBs successfully penetrated the blood-brain barrier and effectively inhibited glioblastoma growth under the combined effects of focused ultrasound and magnetic stimulation. These results provide a new approach for the treatment of glioblastoma, with implications for future clinical translation.

Characteristics of interbody bone graft fusion after transforaminal lumbar interbody fusion according to intervertebral space division.

Background: According to intervertebral space division, the characteristics of interbody bone graft fusion after transforaminal lumbar interbody fusion (TLIF) were assessed via computed tomography (CT) scan to provide a theoretical basis for selecting the bone grafting site of interbody fusion. Methods: The medical records of 57 patients with lumbar spinal stenosis and disc herniation treated with TLIF were analysed retrospectively. In total, 57 segments received lumbar interbody fusion. A thin-layer CT scan was performed to evaluate fusion in each zone of the fusion space. Results: The fusion rates were 57.89% (n = 33) in the anterior cage zone, 73.68% (n = 42) in the posterior cage zone, 66.67% (n = 38) in the decompression zone, 26.32% (n = 15) in the contralateral decompression zone and 94.74% (n = 54) in the inner cage zone. There were significant differences among the fusion rates of the five zones (P < 0.001). Further pairwise comparison revealed that the fusion rates in the inner cage significantly differed from the anterior and posterior cages and decompression and contralateral decompression zones (P = 0.001, 0.002, 0.001 and 0.001, respectively). Conclusion: We think the central cage zone (i.e., inner cage) should be the focus of bone grafting. Although there is small volume of bone graft on the posterior cage zone, the fusion rate is relatively high, only secondary to the inner cage zone. The fusion rate is of the contralateral decompression zone is lower although there is a bone graft.

Riluzole Promotes Neurite Growth in Rats after Spinal Cord Injury through the GSK-3ß/CRMP-2 Pathway.

Spinal cord injury (SCI) is a disastrous event that often leads to permanent neurological deficits involving motor, sensory, and autonomic dysfunctions in patients. Accumulating research has demonstrated that riluzole may play crucial roles in the process of spinal tissue repair, but the underlying mechanisms remain elusive. This study verified the effectiveness of riluzole and speculated that a riluzole-afforded protection mechanism may be associated with the glycogen synthase kinase-3 beta (GSK-3ß)/collapsin response mediator protein-2 (CRMP-2) pathway in rats after spinal cord injury. Here, a modified Allen's weight dropping model was generated and riluzole at 4 mg/kg was injected intraperitoneally after surgery and twice a day for 7 consecutive days. At 6 weeks after SCI, we found that riluzole treatment reduced the central cavity size of the spinal cord and improved neurological functions. Meanwhile, riluzole-treated rats exhibited shorter latency and larger amplitude in motor evoked potentials and somatosensory evoked potentials, compared with vehicle-treated rats. Furthermore, Western blotting and immunofluorescence data revealed that the expression levels of GSK-3ß and phosphorylated-GSK-3ß were lower in riluzole-treated SCI rats compared with vehicle-treated rats. We next detected the expression CRMP-2 and phosphorylated CRMP-2 and found that the expression of CRMP-2 showed no difference between the riluzole-treated and vehicle-treated groups; however, administration of riluzole downregulated phosphorylated CRMP-2 expression. The current findings suggest that after SCI, administration of riluzole promotes neurological functional restoration, which may be associated, in part, with its activation of the GSK-3ß/CRMP-2 signaling pathway.

The Roles of Blood Lipid-Metabolism Genes in Immune Infiltration Could Promote the Development of IDD.

Objectives: Intervertebral disc degeneration is a progressive and chronic disease, usually manifesting as low back pain. This study aimed to screen effective biomarkers for medical practice as well as figuring out immune infiltration situations between circulation and intervertebral discs. Methods: Gene expression profiles of GSE124272 was included for differentially analysis, WGCNA and immune infiltration analysis from GEO database, and other GSE series were used as validation datasets. A series of validation methods were conducted to verify the robustness of hub genes, such as principal component analysis, machine learning models, and expression verification. Lastly, nomogram was established for medical practice. Results: 10 genes were commonly screened via combination of DEGs, WGCNA analysis and lipid metabolism related genes. Furthermore, 3 hub gens CYP27A1, FAR2, CYP1B1 were chosen for subsequent analysis based on validation of different methods. GSEA analysis discovered that neutrophil extracellular traps formation and NOD-like receptor signaling pathway was activated during IDD. Immune infiltration analysis demonstrated that the imbalance of neutrophils and γδT cells were significantly correlated with IDD progression. Nomogram was established based on CYP27A1, FAR2, CYP1B1 and age, the calibration plot confirmed the stability of our model. Conclusion: CYP27A1, FAR2, CYP1B1 were considered as hub lipid metabolism related genes (LMRGs) in the development of IDD, which were regarded as candidate diagnostic biomarkers especially in circulation. The effects are worth expected in the early diagnosis of IDD through detecting these genes in blood.

The impact of intravertebral cleft on cement leakage in percutaneous vertebroplasty for osteoporotic vertebral compression fractures: a case-control study.

BACKGROUND: The impact of intravertebral cleft (IVC) on cement leakage in percutaneous vertebroplasty (PVP) for osteoporotic vertebral compression fractures (OVCFs) has been discussed. However, the results were conflicting, as the study population and cement leakage classification were heterogeneous. The aim of the study was to evaluate the impact of IVC on the incidence of leakage through vein, leakage through cortex as well as general leakage in PVP for OVCFs. METHODS: All patients with OVCFs who underwent PVP between January 2016 and June 2019 at our institution were retrospectively reviewed. Patients were eligible for this case-control study if they were diagnosed as single level fracture in spine. After inclusive and exclusive criteria were met, a total of 139 patients with IVC were enrolled as the study group. Non-IVC controls were matched in a 1:1 ratio in age (within 3 years), sex and fracture severity with patients in study group. Cement leakage were classified into four types [type B (through basivertebral vein), type S (through segmental vein), type-C (through a cortical defect), and type D (intradiscal leakage)], furtherly into two types [venous type (type-B or/and type S) and cortical type (type-C or/and type-D)]. A general leakage rate and a specific leakage rate per each type were compared between both groups. RESULTS: Each group included 139 patients. Groups were homogenous for age, sex, fracture severity, fracture location, fracture type, cement volume, puncture approach and property of cement. Compared with control group, IVC group had a significantly lower rate of type-B (20.9% vs. 31.7%, P = 0.041), type-S (24.5% vs. 52.5%, P = 0.000), and venous type leakage (37.4% vs. 67.6%, P = 0.000), a significantly higher rate of type-C (25.9% vs. 12.2%, P = 0.004), type-D (16.5% vs. 6.5%, P = 0.009), and cortical type leakage (40.3% vs. 16.5%, P = 0.000), no significant difference on the rate of general leakage (67.6% vs. 76.3%, P = 0.109). CONCLUSION: IVC decreased the risk of cement leakage through vein and increased the risk of cement leakage through cortex. However, it had no significant effect on the occurrence of general leakage.

Exosomes Immunity Strategy: A Novel Approach for Ameliorating Intervertebral Disc Degeneration.

Low back pain (LBP), which is one of the most severe medical and social problems globally, has affected nearly 80% of the population worldwide, and intervertebral disc degeneration (IDD) is a common musculoskeletal disorder that happens to be the primary trigger of LBP. The pathology of IDD is based on the impaired homeostasis of catabolism and anabolism in the extracellular matrix (ECM), uncontrolled activation of immunologic cascades, dysfunction, and loss of nucleus pulposus (NP) cells in addition to dynamic cellular and biochemical alterations in the microenvironment of intervertebral disc (IVD). Currently, the main therapeutic approach regarding IDD is surgical intervention, but it could not considerably cure IDD. Exosomes, extracellular vesicles with a diameter of 30-150 nm, are secreted by various kinds of cell types like stem cells, tumor cells, immune cells, and endothelial cells; the lipid bilayer of the exosomes protects them from ribonuclease degradation and helps improve their biological efficiency in recipient cells. Increasing lines of evidence have reported the promising applications of exosomes in immunological diseases, and regarded exosomes as a potential therapeutic source for IDD. This review focuses on clarifying novel therapies based on exosomes derived from different cell sources and the essential roles of exosomes in regulating IDD, especially the immunologic strategy.

Riluzole improves functional recovery after acute spinal cord injury in rats and may be associated with changes in spinal microglia/macrophages polarization.

Spinal cord injury (SCI) triggers pronounced inflammatory responses that are accompanied by neuronal disruption and functional deficits. SCI treatment remains an unmet clinical need. Emerging evidence suggests that riluzole may exert a neuroprotective effect due to its anti-inflammatory properties. However, details of the underlying mechanisms remain poorly defined. The polarization of microglial/macrophages has an important role in neuroinflammation. Here, we examined whether riluzole can exert a neuroprotective effect after acute SCI, and whether this effect is associated with changes in microglia/macrophages polarization. Riluzole (4 mg/kg) or vehicle were injected intraperitoneally (i.p.) in female rats immediately following SCI and repeated for 7 consecutive days (b.i.d.). Compared with vehicle treatment, riluzole-treated SCI rats showed significant higher locomotor scores (Basso, Beattie, and Bresnahan score, Inclined Plane test score, n = 18/group). Riluzole-treated rats also developed smaller spinal cavities, showed higher levels of myelin basic protein (MBP) and neurofilament (NF)200 immunoreactivities, and lower levels of proinflammatory cytokines in the spinal cord at 7 days post-SCI. Immunofluorescence study revealed more CD206+ cells and less iNOS+ cells in the injured spinal cord of riluzole-treated SCI rats, as compared to vehicle control. Using real-time PCR, we found that riluzole upregulated the mRNA levels of M2 markers, but downregulated that of M1 markers, as compared to the vehicle treatment. Current findings suggest that systemic administration of riluzole after acute SCI facilitated motor function recovery and inhibited inflammatory responses, which may be associated with polarization of M2 microglia/macrophages.

[Radiological characteristics and clinical manifestation of isolated lumbar foraminal stenosis].

OBJECTIVE: To discuss radiological characteristics and clinical manifestation of isolated lumbar foraminal stenosis. METHODS: From March 2011 to March 2014, 21 patients with isolated degenerative lumbar foraminal stenosis accepted lumbar decompression and fusion in Beijing Luhe Hospital. Intervertebral disc space was evaluated by measuring the position of joint-body line on preoperative X-ray. Bilateral foraminal area of the corresponding segment in CT (sagittal view of 2D reconstruction) and MRI (T2W1 sagittal view) were measured by Surgimap software. For patients with unilateral symptoms, foraminal area of the affected side was compared with that of the contralateral side. Foraminal area of the same segment on CT was also compared with that on MRI. Preoperatively and at the final follow-up, visual analogue score (VAS) and Oswestry Disability Index (ODI) were used to evaluate clinical outcomes. RESULTS: All patients had a follow-up over 6 months and the average follow-up was 16.8 months (7-42 months). Of the 21 patients (26 segments), 12 segments showed gross narrowing and 14 segments showed slight narrowing. After preoperative measurement on MRI, 6 patients had foraminal stenosis of grade 2, and 15 patients had foraminal stenosis of grade 3, showing no significant difference in clinical outcomes. Compared with the foraminal area of the unaffected side, the affected side showed a decrease of 16% on CT and 28% on MRI, and the difference was statistically significant (t = 3.453, P < 0.05). The foraminal area measured on CT was larger than that measured on MRI (P < 0.05). Compared with that preoperatively, VAS (back pain), VAS (leg pain) and ODI showed significant improvement at the final follow-up (P < 0.05). CONCLUSIONS: Radiological examinations as X-ray, CT, MRI and intervertebral foramen block technique play an important role in the diagnosis of foraminal stenosis. Soft oppression caused by hyperplasia and hypertrophy of transforaminal ligment or joint capsule may be important promoters of degenerative lumbar foraminal stenosis. Lumbar foraminal decompression and interbody fusion can satisfactorily improve preoperative symptoms.

Injury and regeneration of intramuscular connective tissue subjected to various regimes of distraction.

OBJECTIVE: To investigate the effect on intramuscular connective tissue and passive range of joint motion by the stress produced in limb lengthening. METHODS: An animal model of limb lengthening was established in the tibia of rabbits. Distraction was initiated at a rate of 1 mm/d and 2 mm/d in two steps respectively, and both proceeded until 10% and 20% of the tibia length was achieved. Muscle samples were harvested at the time when distraction ended and at the 4th week of consolidation after the distraction. Scanning electron microscope was applied to observe the morphological changes of the perimysium. The goniometer, which we made for this study, was used to measure the passive range of joint motion. RESULTS: The collagen fibers were partitioned in bundles, crimped and interconnected closely and orderly. In the regime of 1 mm/d distraction with 10% lengthening, no apparent changes of the collagen fiber and passive range of joint motion was demonstrated. When tibia was increased to 20%, the crimped fibers showed a tendency of being straightened while the passive range of joint motion was reduced. The findings remained the same at the 4th week of consolidation. In the regime of 2 mm/d distraction with 10% lengthening, the crimped structure of the collagen fibers in the perimysium disappeared and the fibers were almost straightened. Additionally, the interconnection of the collagen fibers became loosened and interstice was presented among the fibers. At the 4th week of consolidation, the restoration to the original crimped structure was not completed. When the lengthening ratio was increased to 20%, the collagen fibers were straightened completely. This condition remained unchanged throughout all 4 weeks. The passive range of joint motion was reduced dramatically in the regime of 2 mm/d distraction. CONCLUSION: The ultrastructure of perimysium and the passive range of joint motion in the regime of 1mm/d lengthening shows the condition closest to the normal ones. The regime of 2 mm/d lengthening may cause an apparent change in the ultrastructure of perimysium and passive range of joint motion.