Mechanically robust and personalized silk fibroin-magnesium composite scaffolds with water-responsive shape-memory for irregular bone regeneration.

The regeneration of critical-size bone defects, especially those with irregular shapes, remains a clinical challenge. Various biomaterials have been developed to enhance bone regeneration, but the limitations on the shape-adaptive capacity, the complexity of clinical operation, and the unsatisfied osteogenic bioactivity have greatly restricted their clinical application. In this work, we construct a mechanically robust, tailorable and water-responsive shape-memory silk fibroin/magnesium (SF/MgO) composite scaffold, which is able to quickly match irregular defects by simple trimming, thus leading to good interface integration. We demonstrate that the SF/MgO scaffold exhibits excellent mechanical stability and structure retention during the degradative process with the potential for supporting ability in defective areas. This scaffold further promotes the proliferation, adhesion and migration of osteoblasts and the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro. With suitable MgO content, the scaffold exhibits good histocompatibility, low foreign-body reactions (FBRs), significant ectopic mineralisation and angiogenesis. Skull defect experiments on male rats demonstrate that the cell-free SF/MgO scaffold markedly enhances bone regeneration of cranial defects. Taken together, the mechanically robust, personalised and bioactive scaffold with water-responsive shape-memory may be a promising biomaterial for clinical-size and irregular bone defect regeneration.

Single-cell sequencing technology in diabetic wound healing: New insights into the progenitors-based repair strategies.

Diabetes mellitus (DM), an increasingly prevalent chronic metabolic disease, is characterised by prolonged hyperglycaemia, which leads to long-term health consequences. Although much effort has been put into understanding the pathogenesis of diabetic wounds, the underlying mechanisms remain unclear. The advent of single-cell RNA sequencing (scRNAseq) has revolutionised biological research by enabling the identification of novel cell types, the discovery of cellular markers, the analysis of gene expression patterns and the prediction of developmental trajectories. This powerful tool allows for an in-depth exploration of pathogenesis at the cellular and molecular levels. In this editorial, we focus on progenitor-based repair strategies for diabetic wound healing as revealed by scRNAseq and highlight the biological behaviour of various healing-related cells and the alteration of signalling pathways in the process of diabetic wound healing. ScRNAseq could not only deepen our understanding of the complex biology of diabetic wounds but also identify and validate new targets for intervention, offering hope for improved patient outcomes in the management of this challenging complication of DM.

Evaluation of genetic response of mesenchymal stem cells to nanosecond pulsed electric fields by whole transcriptome sequencing.

BACKGROUND: Mesenchymal stem cells (MSCs) modulated by various exogenous signals have been applied extensively in regenerative medicine research. Notably, nanosecond pulsed electric fields (nsPEFs), characterized by short duration and high strength, significantly influence cell phenotypes and regulate MSCs differentiation via multiple pathways. Consequently, we used transcriptomics to study changes in messenger RNA (mRNA), long noncoding RNA (lncRNA), microRNA (miRNA), and circular RNA expression during nsPEFs application. AIM: To explore gene expression profiles and potential transcriptional regulatory mechanisms in MSCs pretreated with nsPEFs. METHODS: The impact of nsPEFs on the MSCs transcriptome was investigated through whole transcriptome sequencing. MSCs were pretreated with 5-pulse nsPEFs (100 ns at 10 kV/cm, 1 Hz), followed by total RNA isolation. Each transcript was normalized by fragments per kilobase per million. Fold change and difference significance were applied to screen the differentially expressed genes (DEGs). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to elucidate gene functions, complemented by quantitative polymerase chain reaction verification. RESULTS: In total, 263 DEGs were discovered, with 92 upregulated and 171 downregulated. DEGs were predominantly enriched in epithelial cell proliferation, osteoblast differentiation, mesenchymal cell differentiation, nuclear division, and wound healing. Regarding cellular components, DEGs are primarily involved in condensed chromosome, chromosomal region, actin cytoskeleton, and kinetochore. From aspect of molecular functions, DEGs are mainly involved in glycosaminoglycan binding, integrin binding, nuclear steroid receptor activity, cytoskeletal motor activity, and steroid binding. Quantitative real-time polymerase chain reaction confirmed targeted transcript regulation. CONCLUSION: Our systematic investigation of the wide-ranging transcriptional pattern modulated by nsPEFs revealed the differential expression of 263 mRNAs, 2 miRNAs, and 65 lncRNAs. Our study demonstrates that nsPEFs may affect stem cells through several signaling pathways, which are involved in vesicular transport, calcium ion transport, cytoskeleton, and cell differentiation.

Using a 3D Navigation Template to Increase the Accuracy of Thoracic Pedicle Screws in Patients with Scoliosis.

This study compares the accuracy and safety of pedicle screw placement using a 3D navigation template with the free-hand fluoroscopy technique in scoliotic patients. Fifteen scoliotic patients were recruited and divided into a template group (eight cases) and a free-hand group (seven cases). All patients received posterior corrective surgeries, and the pedicle screw was placed using a 3D navigation template or a free-hand technique. After surgery, the positions of the pedicle screws were evaluated using CT. A total of 264 pedicle screws were implanted in 15 patients. Both the two techniques were found to achieve satisfactory safety of screw insertion in scoliotic patients (89.9% vs. 90.5%). In the thoracic region, the 3D navigation template was able to achieve a much higher accuracy of screw than the free-hand technique (75.3% vs. 60.4%). In the two groups, the accuracy rates on the convex side were slightly higher than on the concave side, while no significance was seen. In terms of rotational vertebrae, no significant differences were seen in Grades I or II vertebrae between the two groups. In conclusion, the 3D navigation template technique significantly increased the accuracy of thoracic pedicle screw placement, which held great potential for extensively clinical application.

Identifying Intraoperative Spinal Cord Injury Location from Somatosensory Evoked Potentials' Time-Frequency Components.

Excessive distraction in corrective spine surgery can lead to iatrogenic distraction spinal cord injury. Diagnosis of the location of the spinal cord injury helps in early removal of the injury source. The time-frequency components of the somatosensory evoked potential have been reported to provide information on the location of spinal cord injury, but most studies have focused on contusion injuries of the cervical spine. In this study, we established 19 rat models of distraction spinal cord injury at different levels and collected the somatosensory evoked potentials of the hindlimb and extracted their time-frequency components. Subsequently, we used k-medoid clustering and naive Bayes to classify spinal cord injury at the C5 and C6 level, as well as spinal cord injury at the cervical, thoracic, and lumbar spine, respectively. The results showed that there was a significant delay in the latency of the time-frequency components distributed between 15 and 30 ms and 50 and 150 Hz in all spinal cord injury groups. The overall classification accuracy was 88.28% and 84.87%. The results demonstrate that the k-medoid clustering and naive Bayes methods are capable of extracting the time-frequency component information depending on the spinal cord injury location and suggest that the somatosensory evoked potential has the potential to diagnose the location of a spinal cord injury.

Intervertebral Disc Progenitors: Lessons Learned from Single-Cell RNA Sequencing and the Role in Intervertebral Disc Regeneration.

The tremendous personal and economic burden worldwide caused by low back pain (LBP) has been surging in recent years. While intervertebral disc degeneration (IVDD) is the leading cause of LBP and vast efforts have been made to develop effective therapies, this problem is far from being resolved, as most treatments, such as painkillers and surgeries, mainly focus on relieving the symptoms rather than reversing the cause of IVDD. However, as stem/progenitor cells possess the potential to regenerate IVD, a deeper understanding of the early development and role of these cells could help to improve the effectiveness of stem/progenitor cell therapy in treating LBP. Single-cell RNA sequencing results provide fresh insights into the heterogeneity and development patterns of IVD progenitors; additionally, we compare mesenchymal stromal cells and IVD progenitors to provide a clearer view of the optimal cell source proposed for IVD regeneration.

Upregulation of ß-catenin signaling represents a single common pathway leading to the various phenotypes of spinal degeneration and pain.

Spine degeneration is an aging-related disease, but its molecular mechanisms remain unknown, although elevated ß-catenin signaling has been reported to be involved in intervertebral disc degeneration. Here, we determined the role of ß-catenin signaling in spinal degeneration and in the homeostasis of the functional spinal unit (FSU), which includes the intervertebral disc, vertebra and facet joint and is the smallest physiological motion unit of the spine. We showed that pain sensitivity in patients with spinal degeneration is highly correlated with ß-catenin protein levels. We then generated a mouse model of spinal degeneration by transgenic expression of constitutively active ß-catenin in Col2+ cells. We found that ß-catenin-TCF7 activated the transcription of CCL2, a known critical factor in osteoarthritic pain. Using a lumbar spine instability model, we showed that a ß-catenin inhibitor relieved low back pain. Our study indicates that ß-catenin plays a critical role in maintaining spine tissue homeostasis, its abnormal upregulation leads to severe spinal degeneration, and its targeting could be an avenue to treat this condition.

Amelioration of ligamentum flavum hypertrophy using umbilical cord mesenchymal stromal cell-derived extracellular vesicles.

Ligamentum flavum (LF) hypertrophy (LFH) has been recognised as one of the key contributors to lumbar spinal stenosis. Currently, no effective methods are available to ameliorate this hypertrophy. In this study, human umbilical cord mesenchymal stromal cell-derived extracellular vesicles (hUCMSC-EVs) were introduced for the first time as promising vehicles for drug delivery to treat LFH. The downregulation of miR-146a-5p and miR-221-3p expressions in human LF tissues negatively correlated with increased LF thickness. The hUCMSC-EVs enriched with these two miRNAs significantly suppressed LFH in vivo and notably ameliorated the progression of transforming growth factor ß1(TGF-ß1)-induced fibrosis in vitro after delivering these two miRNAs to mouse LF cells. The results further demonstrated that miR-146a-5p and miR-221-3p directly bonded to the 3'-UTR regions of SMAD4 mRNA, thereby inhibiting the TGF-ß/SMAD4 signalling pathway. Therefore, this translational study determined the effectiveness of a hUCMSC-EVs-based approach for the treatment of LFH and revealed the critical target of miR-146a-5p and miR-221-3p. Our findings provide new insights into promising therapeutics using a hUCMSC-EVs-based delivery system for patients with lumbar spinal stenosis.

Annular Defects Impair the Mechanical Stability of the Intervertebral Disc.

STUDY DESIGN: A biomechanical study. OBJECTIVES: The purpose of this study was to investigate the effects of cruciform and square incisions of annulus fibrosus (AF) on the mechanical stability of bovine intervertebral disc (IVD) in multiple degrees of freedom. METHODS: Eight bovine caudal IVD motion segments (bone-disc-bone) were obtained from the local abattoir. Cruciform and square incisions were made at the right side of the specimen's annulus using a surgical scalpel. Biomechanical testing of three-dimensional 6 degrees of freedom was then performed on the bovine caudal motion segments using the mechanical testing and simulation (MTS) machine. Force, displacement, torque and angle were recorded synchronously by the MTS system. P value <.05 was considered statistically significant. RESULTS: Cruciform and square incisions of the AF reduced both axial compressive and torsional stiffness of the IVD and were significantly lower than those of the intact specimens (P < .01). Left-side axial torsional stiffness of the cruciform incision was significantly higher than a square incision (P < .01). Neither incision methods impacted flexional-extensional stiffness or lateral-bending stiffness. CONCLUSIONS: The cruciform and square incisions of the AF obviously reduced axial compression and axial rotation, but they did not change the flexion-extension and lateral-bending stiffness of the bovine caudal IVD. This mechanical study will be meaningful for the development of new approaches to AF repair and the rehabilitation of the patients after receiving discectomy.

Identification of injury type using somatosensory and motor evoked potentials in a rat spinal cord injury model.

The spinal cord is at risk of injury during spinal surgery. If intraoperative spinal cord injury is identified early, irreversible impairment or loss of neurological function can be prevented. Different types of spinal cord injury result in damage to different spinal cord regions, which may cause different somatosensory and motor evoked potential signal responses. In this study, we examined electrophysiological and histopathological changes between contusion, distraction, and dislocation spinal cord injuries in a rat model. We found that contusion led to the most severe dorsal white matter injury and caused considerable attenuation of both somatosensory and motor evoked potentials. Dislocation resulted in loss of myelinated axons in the lateral region of the injured spinal cord along the rostrocaudal axis. The amplitude of attenuation in motor evoked potential responses caused by dislocation was greater than that caused by contusion. After distraction injury, extracellular spaces were slightly but not significantly enlarged; somatosensory evoked potential responses slightly decreased and motor evoked potential responses were lost. Correlation analysis showed that histological and electrophysiological findings were significantly correlated and related to injury type. Intraoperative monitoring of both somatosensory and motor evoked potentials has the potential to identify iatrogenic spinal cord injury type during surgery.

Tumor necrosis factor-α inhibition restores matrix formation by human adipose-derived stem cells in the late stage of chondrogenic differentiation.

BACKGROUND: Cartilage tissue engineering is a promising strategy for treating cartilage damage. Matrix formation by adipose-derived stem cells (ADSCs), which are one type of seed cell used for cartilage tissue engineering, decreases in the late stage of induced chondrogenic differentiation in vitro, which seriously limits research on ADSCs and their application. AIM: To improve the chondrogenic differentiation efficiency of ADSCs in vitro, and optimize the existing chondrogenic induction protocol. METHODS: Tumor necrosis factor-alpha (TNF-α) inhibitor was added to chondrogenic culture medium, and then Western blotting, enzyme linked immunosorbent assay, immunofluorescence and toluidine blue staining were used to detect the cartilage matrix secretion and the expression of key proteins of nuclear factor kappa-B (NF-κB) signaling pathway. RESULTS: In this study, we found that the levels of TNF-α and matrix metalloproteinase 3 were increased during the chondrogenic differentiation of ADSCs. TNF-α then bound to its receptor and activated the NF-κB pathway, leading to a decrease in cartilage matrix synthesis and secretion. Blocking TNF-α with its inhibitors etanercept (1 µg/mL) or infliximab (10 µg/mL) significantly restored matrix formation. CONCLUSION: Therefore, this study developed a combination of ADSC therapy and targeted anti-inflammatory drugs to optimize the chondrogenesis of ADSCs, and this approach could be very beneficial for translating ADSC-based approaches to treat cartilage damage.

Regulated cell death: Implications for intervertebral disc degeneration and therapy.

As a controllable biological process, regulated cell death (RCD) extensively participates in cellular homeostasis, organismal development, and the pathogenesis of diseases. This review addresses the research gaps by synthesising the findings on the complexity of RCD modes and their role in disc degeneration, and summarises the preclinical strategies to alleviate disc degeneration and promote disc repair by regulating RCD. Background: Intervertebral disc degeneration (IDD) is the major source of chronic low back pain. As a controllable biological process, regulated cell death (RCD) extensively participates in the pathogenesis of IDD. Nevertheless, the initiation and progression of RCD remain unclear, and more importantly, the interaction between different RCD modes during IDD and therapy is far from well understood. Methods: Literature search was performed using "regulated cell death AND intervertebral disc degeneration" in PubMed, Embase, and Web of Science. Meanwhile, relevant findings have been reviewed and quoted. Results: In this review, we discuss the inducing factors of IDD, various modes of RCD in intervertebral disc, the interactions between different RCD modes, as well as the obstacles to achieve disc regeneration. Meanwhile, the research gaps and perspective in studies that targeting RCD are also presented. Conclusion: Increasing evidence demonstrated the presence of different RCD modes in intervertebral disc during the progression of IDD. RCD in the resident disc cells is probably induced by the multiple factors such as abnormal mechanical loading, nutritional imbalance, inflammation microenvironment, circadian rhythm changes, withdraw of hormones, and other biomechanical factors. A better understanding of the fundamental mechanisms and the interactions between different RCD modes might contribute to the rescuing of disc degeneration and development of promising therapeutics. Translational potential statement: The Translational potential of this article. This review aims to demonstrate a better understanding of the fundamental mechanisms governing RCD, which might contribute to the rescuing of disc degeneration and to the development of promising therapeutics in a clinical setting.

Is the Zero-P Spacer Suitable for 3-Level Anterior Cervical Discectomy and Fusion Surgery in Terms of Sagittal Alignment Reconstruction: A Comparison Study with Traditional Plate and Cage System.

The Zero-P spacer was primarily developed aiming to reduce the morbidity associated with the traditional anterior cervical plate. During the past decade, many authors have reported the use of Zero-P spacers for anterior cervical discectomy and fusion (ACDF) of one or two segments. Nevertheless, there is still a paucity of knowledge on the safety and feasibility of using Zero-P spacers for 3-level fixation. The objective of this study was to investigate the clinical and radiological outcomes, with a focus on the sagittal alignment reconstruction of 3-level ACDF surgery using Zero-P spacers versus those using a traditional plate and cage system. From Sep 2013 to Aug 2016, a total of 44 patients who received 3-level ACDF surgery due to cervical spondylotic myelopathy were recruited. The Zero-P spacer was used in 23 patients (group ZP) and the traditional plate and cage system in 21 (group PC). Clinical outcomes were analyzed by Neck Disability Index (NDI) and Japanese Orthopedic Association (JOA) scores, and dysphagia was evaluated using the Bazaz score. Radiological outcomes, including fusion rate, adjacent segment degeneration (ASD), and especially changes in cervical sagittal alignment, were analyzed. The NDI and JOA scores did not differ significantly between the two groups postoperatively (p > 0.05); however, there was significantly less dysphagia in patients using Zero-P spacers at the 3- and 6-month follow-up (p < 0.05). At the 24-month follow-up, the fusion rate and ASD were similar between the two groups (p > 0.05). Interestingly, patients using Zero-P spacers had a significantly lower postoperative C2-7 Cobb angle and fused segment Cobb angle, compared to those using a traditional plate and cage system (p < 0.05); meanwhile, the fused segment disc wedge was also found to be significantly smaller in patients using Zero-P spacers after surgery (p < 0.05). Moreover, we further divided patients into subgroups according to their cervical lordosis. In patients with a preoperative C2-7 Cobb angle ≤ 10°, significantly less cervical and local lordosis, as well as disc wedge, were seen in group ZP after surgery (p < 0.05), while in others with a preoperative C2-7 Cobb angle > 10°, no significant difference in postoperative changes of the cervical sagittal alignment was seen between group ZP and group PC (p > 0.05). Zero-P spacers used in 3-level ACDF surgery could provide equivalent clinical outcomes and a lower rate of postoperative dysphagia, compared to the traditional plate and cage system. However, our results showed that it was inferior to the cervical plate in terms of sagittal alignment reconstruction for 3-level fixation. We recommend applying Zero-P spacers for 3-level ACDF in patients with good preoperative cervical lordosis (C2-7 Cobb angle > 10°), in order to restore and maintain physiological curvature of the cervical spine postoperatively.

Species variation in the cartilaginous endplate of the lumbar intervertebral disc.

Backgrounds: Cartilaginous endplate (CEP) plays an essential role in intervertebral disc (IVD) health and disease. The aim was to compare the CEP structure of lumbar IVD and to reveal the detailed pattern of integration between the CEP and bony endplate (BEP) from different species. Methods: A total of 34 IVDs (5 human, 5 goat, 8 pig, 8 rabbit, and 8 rat IVDs) were collected, fixed and midsagittally cut; in each IVD, one-half was used for histological staining to observe the CEP morphology, and the other half was used for scanning electron microscopy (SEM) analysis to measure the diameters and distributions of collagen fibers in the central and peripheral CEP areas and to observe the pattern of CEP-BEP integration from different species. Results: The human, pig, goat, and rabbit IVDs had the typical BEP-CEP structure, but the rat CEP was directly connected with the growth plate. Human CEP was the thickest (896.95 ± 87.71 µm) among these species, followed by pig, goat, rat, and rabbit CEPs. Additionally, the mean cellular density of the rabbit CEP was the highest, which was 930 ± 202 per mm2, followed by the rat, goat, pig, and human CEPs. In all the species, the collagen fiber diameter in the peripheral area was much bigger than that in the central area. The collagen fiber diameters of CEP from the human, pig, goat, and rat were distributed between 35 nm and 65 nm. The BEP and CEP were connected by the collagen from the CEP, aggregating into bundles or cross links with each other to form a network, and anchored to BEP. Conclusions: Significant differences in the thickness, cellular density, and collagen characterization of CEPs from different species were demonstrated; the integration of BEP-CEP in humans, pigs, goats, and rabbits was mainly achieved by the collagen bundles anchoring system, while the typical BEP-CEP interface did not exist in rats.

[Comparison of short-term effectiveness of Prodisc-C Vivo artificial disc replacement and Zero-P fusion for treatment of single-segment cervical spondylosis].

Objective: To compare the short-term effectiveness and the impact on cervical segmental range of motion using Prodisc-C Vivo artificial disc replacement and Zero-P fusion for the treatment of single-segment cervical spondylosis. Methods: The clinical data of 56 patients with single-segment cervical spondylosis who met the selection criteria between January 2015 and December 2018 were retrospectively analyzed, and they were divided into study group (27 cases, using Prodisc-C Vivo artificial disc replacement) and control group (29 cases, using Zero-P fusion) according to different surgical methods. There was no significant difference between the two groups in terms of gender, age, type of cervical spondylosis, disease duration, involved segments and preoperative pain visual analogue scale (VAS) score, Japanese Orthopaedic Association (JOA) score, neck disability index (NDI), surgical segments range of motion, upper and lower adjacent segments range of motion, overall cervical spine range of motion, and cervical curvature (P>0.05). The operation time, intraoperative blood loss, postoperative hospitalization stay, time of returning to work, clinical effectiveness indicators (VAS score, JOA score, NDI, and improvement rate of each score), and imaging indicators (surgical segments range of motion, upper and lower adjacent segments range of motion, overall cervical spine range of motion, and cervical curvature, prosthesis position, bone absorption, heterotopic ossification, etc.) were recorded and compared between the two groups. Results: There was no significant difference in operation time and intraoperative blood loss between the two groups (P>0.05); the postoperative hospitalization stay and time of returning to work in the study group were significantly shorter than those in the control group (P<0.05). Both groups were followed up 12-64 months, with an average of 26 months. There was no complication such as limb or organ damage, implant failure, and severe degeneration of adjacent segments requiring reoperation. The VAS score, JOA score, and NDI of the two groups at each time point after operation significantly improved when compared with those before operation (P<0.05); there was no significant difference in the above scores at each time point after operation between the two groups (P>0.05); there was no significant difference in the improvement rate of each score between the two groups at last follow-up (P>0.05). The surgical segments range of motion in the study group maintained to varying degrees after operation, while it in the control group basically disappeared after operation, showing significant differences between the two groups (P<0.05). At last follow-up, there was no significant difference in the upper and lower adjacent segments range of motion in the study group when compared with preoperative ones (P>0.05), while the upper adjacent segments range of motion in the control group increased significantly (P<0.05). The overall cervical spine range of motion and cervical curvature of the two groups decreased at 3 months after operation, and increased to varying degrees at last follow-up, but there was no significant difference between groups and within groups (P>0.05). At last follow-up, X-ray films and CT examinations showed that no prosthesis loosening, subsidence, or displacement was found in all patients; there were 2 cases (7.4%) of periprosthetic bone resorption and 3 cases (11.1%) of heterotopic ossification which did not affect the surgical segments range of motion. Conclusion: Both the Prodisc-C Vivo artificial disc replacement and Zero-P fusion have satisfactory short-term effectiveness in treatment of single-segment cervical spondylosis. Prodisc-C Vivo artificial disc replacement can also maintain the cervical spine range of motion to a certain extent, while reducing the occurrence of excessive motion of adjacent segments after fusion.

Different Time-Frequency Distribution Patterns of Somatosensory Evoked Potentials in Dual- and Single-Level Spinal Cord Compression.

Among patients with cervical myelopathy, the most common level of stenosis at spinal cord of all ages was reported to be between cervical levels C5-6. Previous studies found that time-frequency components (TFCs) of somatosensory evoked potentials (SEPs) possess location information of spinal cord injury (SCI) in single-level deficits in the spinal cord. However, the clinical reality is that there are multiple compressions at multiple spinal cord segments. This study proposed a new algorithm to differentiate distribution patterns of SEP TFCs between the dual-level compression and the corresponding single-level compression, which is potential in providing precise diagnosis of cervical myelopathy. In the present animal study, a group of rats with dual-level compressive (C5+6) injury to cervical spinal cord was investigated. SEPs were collected at 2 weeks after surgery, while SEP TFCs were calculated. The SEP TFCs under dual-level compression were compared to an existent dataset with one sham control group and three single level compression groups at C4, C5, C6. Behavioral evaluation showed very similar scale of injury severity between individual rats, while histology evaluation confirmed the precise location of injury. According to time-frequency distribution patterns, it showed that the middle-energy components of dual-level showed similar patterns as that of each single-level group. In addition, the low-energy components of the dual-level C5+6 group had the highest correlation with C5 (R = 0.3423, p < 0.01) and C6 (R = 0.4000, p < 0.01) groups, but much lower with C4 group (R = 0.1071, p = 0.012). These results indicated that SEP TFCs components possess information regarding the location of neurological lesion after spinal cord compression. It preliminarily demonstrated that SEP TFCs are likely a useful measure to provide location information of neurological lesions after compression SCI.

Membranous Extracellular Matrix-Based Scaffolds for Skin Wound Healing.

Membranous extracellular matrix (ECM)-based scaffolds are one of the most promising biomaterials for skin wound healing, some of which, such as acellular dermal matrix, small intestinal submucosa, and amniotic membrane, have been clinically applied to treat chronic wounds with acceptable outcomes. Nevertheless, the wide clinical applications are always hindered by the poor mechanical properties, the uncontrollable degradation, and other factors after implantation. To highlight the feasible strategies to overcome the limitations, in this review, we first outline the current clinical use of traditional membranous ECM scaffolds for skin wound healing and briefly introduce the possible repair mechanisms; then, we discuss their potential limitations and further summarize recent advances in the scaffold modification and fabrication technologies that have been applied to engineer new ECM-based membranes. With the development of scaffold modification approaches, nanotechnology and material manufacturing techniques, various types of advanced ECM-based membranes have been reported in the literature. Importantly, they possess much better properties for skin wound healing, and would become promising candidates for future clinical translation.

Role of the miR-133a-5p/FBXO6 axis in the regulation of intervertebral disc degeneration.

OBJECTIVE: Low back pain is a leading cause of disabilities worldwide, and intervertebral disc degeneration (IVDD)-related disorders have been recognised as one of the main contributors. Nevertheless, the underlying mechanism has not yet been fully understood. The aim of this study was to investigate the role of the miR-133a-5p/FBXO6 axis in the regulation of IVDD. METHODS: RT-qPCR, WB and IHC were performed to assess the expression of FBXO6 in human IVD tissues. Nucleus pulposus (NP) cells were treated with IL-1ß to induce IVDD cellular model. Silence of FBXO6 was achieved using specific siRNAs. CCK-8 assay, flow cytometry, TUNEL assay, RT-qPCR and WB were used to evaluate the role and mechanism of FBXO6 in the process of IVDD. Online tools, GSE datasets and RT-qPCR were used to search the candidate miRNAs targeting FBXO6. The direct binding sites between FBXO6 and miR-133a-5p were further verified by a dual luciferase assay. RT-qPCR, WB and rescue experiments were conducted to identify the regulatory function of miR-133a-5p on the expression of aggrecan, collagen Ⅱ, MMP3, ADAMTS5, IL-6 and COX2. In addition, the role of the NF-κB pathway in regulating miR-133a-5p was studied using lentiviral shRNA, WB and RT-qPCR. RESULTS: Results showed that FBXO6 mainly expressed in the NP tissue of IVD and the expression of FBXO6 decreased with the process of IVDD as well as under IL-1ß stimulation. The silence of FBXO6 led to the decreased expression of aggrecan and collagen Ⅱ and the increased expression of MMP3, ADAMTS5, IL-6 and COX2, which further induced the degeneration of NP cells. The bioinformatic analysis showed that miR-133a-5p was the candidate miRNA targeting FBXO6. miR-133a-5p was upregulated in IVDD tissues and significantly inhibited the expression of FBXO6. The inhibition of miR-133a-5p ameliorated the acceleration of IVDD induced by the silence of FBXO6 in vitro. Moreover, it was demonstrated that IL-1ß regulated the expression of the miR-133a-5p/FBXO6 axis via the NF-κB pathway in NP cells. CONCLUSION: miR-133a-5p was upregulated by IL-1ß to aggravate intervertebral disc degeneration via sponging FBXO6. Inhibiting miR-133a-5p expression or rescuing FBXO6 expression may be promising strategies for the treatment of IVDD. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: This study suggests that the miR-133a-5p/FBXO6 axis could regulate NP cells proliferation, apoptosis, synthesis and degradation of extracellular matrix, which provides a promising therapeutic target and strategy for the treatment of IVDD.

Lithium and Copper Induce the Osteogenesis-Angiogenesis Coupling of Bone Marrow Mesenchymal Stem Cells via Crosstalk between Canonical Wnt and HIF-1α Signaling Pathways.

The combination of osteogenesis and angiogenesis dual-delivery trace element-carrying bioactive scaffolds and stem cells is a promising method for bone regeneration and repair. Canonical Wnt and HIF-1α signaling pathways are vital for BMSCs' osteogenic differentiation and secretion of osteogenic factors, respectively. Simultaneously, lithium (Li) and copper (Cu) can activate the canonical Wnt and HIF-1α signaling pathway, respectively. Moreover, emerging evidence has shown that the canonical Wnt and HIF signaling pathways are related to coupling osteogenesis and angiogenesis. However, it is still unclear whether the lithium- and copper-doped bioactive scaffold can induce the coupling of the osteogenesis and angiogenesis in BMSCs and the underlying mechanism. So, we fabricated a lithium- (Li+-) and copper- (Cu2+-) doped organic/inorganic (Li 2.5-Cu 1.0-HA/Col) scaffold to evaluate the coupling osteogenesis and angiogenesis effects of lithium and copper on BMSCs and further explore its mechanism. We investigated that the sustained release of lithium and copper from the Li 2.5-Cu 1.0-HA/Col scaffold could couple the osteogenesis- and angiogenesis-related factor secretion in BMSCs seeding on it. Moreover, our results showed that 500 µM Li+ could activate the canonical Wnt signaling pathway and rescue the XAV-939 inhibition on it. In addition, we demonstrated that the 25 µM Cu2+ was similar to 1% oxygen environment in terms of the effectiveness of activating the HIF-1α signaling pathway. More importantly, the combination stimuli of Li+ and Cu2+ could couple the osteogenesis and angiogenesis process and further upregulate the osteogenesis- and angiogenesis-related gene expression via crosstalk between the canonical Wnt and HIF-1α signaling pathway. In conclusion, this study revealed that lithium and copper could crosstalk between the canonical Wnt and HIF-1α signaling pathways to couple the osteogenesis and angiogenesis in BMSCs when they are sustainably released from the Li-Cu-HA/Col scaffold.

Modified surgical treatment for a patient with neurofibromatosis scoliosis: A case report.

BACKGROUND: The correction surgery for severely multidimensional spinal deformity in neurofibromatosis type I is very difficult and it is still a very big challenge for spine surgeons. CASE SUMMARY: A 44-year-old woman presented with progressive kyphosis for more than 10 years and low back pain for 2 years. She had been diagnosed with neurofibromatosis at a local hospital many years ago. Conservative treatments had been applied, but the symptoms got worse rather than alleviated. Therefore, surgery was required. CONCLUSION: For this patient with severe deformity, the correction treatment of Ponte osteotomy followed by satellite rod technique in the region of the apical vertebra and the technique of pedicle screws and dual iliac screws had been applied, and successful clinical outcomes were achieved.