The treatment efficacy of bone tissue engineering strategy for repairing segmental bone defects under diabetic condition.

Background: Diabetes mellitus is a systematic disease which exert detrimental effect on bone tissue. The repair and reconstruction of bone defects in diabetic patients still remain a major clinical challenge. This study aims to investigate the potential of bone tissue engineering approach to improve bone regeneration under diabetic condition. Methods: In the present study, decalcified bone matrix (DBM) scaffolds were seeded with allogenic fetal bone marrow-derived mesenchymal stem cells (BMSCs) and cultured in osteogenic induction medium to fabricate BMSC/DBM constructs. Then the BMSC/DBM constructs were implanted in both subcutaneous pouches and large femoral bone defects in diabetic (BMSC/DBM in DM group) and non-diabetic rats (BMSC/DBM in non-DM group), cell-free DBM scaffolds were implanted in diabetic rats to serve as the control group (DBM in DM group). X-ray, micro-CT and histological analyses were carried out to evaluate the bone regenerative potential of BMSC/DBM constructs under diabetic condition. Results: In the rat subcutaneous implantation model, quantitative micro-CT analysis demonstrated that BMSC/DBM in DM group showed impaired bone regeneration activity compared with the BMSC/DBM in non-DM group (bone volume: 46 ± 4.4 mm3 vs 58.9 ± 7.15 mm3, *p < 0.05). In the rat femoral defect model, X-ray examination demonstrated that bone union was delayed in BMSC/DBM in DM group compared with BMSC/DBM in non-DM group. However, quantitative micro-CT analysis showed that after 6 months of implantation, there was no significant difference in bone volume and bone density between the BMSC/DBM in DM group (199 ± 63 mm3 and 593 ± 65 mg HA/ccm) and the BMSC/DBM in non-DM group (211 ± 39 mm3 and 608 ± 53 mg HA/ccm). Our data suggested that BMSC/DBM constructs could repair large bone defects in diabetic rats, but with delayed healing process compared with non-diabetic rats. Conclusion: Our study suggest that biomaterial sacffolds seeded with allogenic fetal BMSCs represent a promising strategy to induce and improve bone regeneration under diabetic condition.

Protocol for optimized dissociation of human scalp tissue for hair follicle transcriptomics by scRNA-seq.

Single-cell RNA sequencing (scRNA-seq) is a powerful tool for studying transcriptomics. Here, we present an optimized protocol for dissociating human scalp tissue and acquiring high-quality single-cell suspension for scRNA-seq to study transcriptomics of human hair follicles. We describe steps for human scalp tissue cleaning, subcutaneous fat removal, mechanical mincing, and enzymatic digestion. We then detail procedures for cleaning, resuspending, a cell viability assay, and library construction.

Clinical Observation of Posterior Approach for Surgical Treatment of Thoracolumbar Pseudarthrosis in Ankylosing Spinal Disorders.

OBJECTIVE: The objective of this study was to evaluate the surgical effectiveness of posterior procedure with long segment stabilization for treating thoracolumbar pseudarthrosis associated with ankylosing spinal disorders (ASDs) without anterior fusion or osteotomy. METHODS: Twelve patients with thoracolumbar pseudarthrosis in ASD were enrolled. All patients underwent posterior long-segment stabilization procedures. In some patients, the percutaneous technique or the aid of a robot or O-arm navigation was utilized for pedicle screw implantation. The clinical results were evaluated by means of the visual analog scale and Oswestry Disability Index. Radiological outcomes were evaluated for bone fusion, anterior column defect, local kyphotic correction, and position of the pedicle screws. RESULTS: All patients experienced effective bone fusion at the sites of pseudarthrosis. The mean operative time was 161.7 ± 57.1 minutes, and the average amount of blood loss was 305.8 ± 293.2 mL. For 6 patients who underwent surgery with the assistance of a robot or O-arm navigation, there was no statistically significant difference observed in terms of operative time and mean blood loss compared to those who used the freehand technique (P > 0.05). The visual analog scale score, Oswestry Disability Index value, and mean local kyphotic angle showed significant improvements at the final follow-up (P < 0.05). The accuracy of pedicle screw placement was 96%. CONCLUSIONS: Posterior surgery with long-segment fixation, without anterior fusion or osteotomy, can achieve satisfactory outcomes in ASD patients with thoracolumbar pseudarthrosis. The application of percutaneous techniques, as well as the assistance of robots or navigation technique may be a good choice for the treatment of pseudarthrosis in ASD patients.

Initial experience of 3-dimensional exoscope in decompression of massive lumbar disc herniation.

OBJECTIVES: To investigate the effect of a three-dimensional (3D) exoscope for decompression of single-segment massive lumbar disc herniation (LDH). METHODS: The study included 56 consecutive patients with single segment massive LDH who underwent decompression assisted by a 3D exoscope from October 2019 to October 2022 at a university hospital. The analysis was based on comparison of perioperative metrics including decompression time, estimated blood loss (EBL) during decompression and postoperative length of stay (PLS); clinical outcomes including assessment using the visual analogue scale (VAS) and the Oswestry disability index (ODI); and incidence of reoperation and complications. RESULTS: The mean decompression time was 28.35 ± 8.93 min (lumbar interbody fusion (LIF)) and 15.50 ± 5.84 min (fenestration discectomy (LOVE surgery)), the mean EBL during decompression was 42.65 ± 12.42 ml (LIF) and 24.32 ± 8.61 ml (LOVE surgery), and the mean PLS was 4.56 ± 0.82 days (LIF) and 2.00 ± 0.65 days (LOVE surgery). There were no complications such as cerebrospinal fluid leakage, nerve root injury and epidural hematoma. All patients who underwent decompression assisted by a 3D exoscope were followed up for 6 months. At the last follow-up, the VAS and ODI scores were significantly improved from the preoperative period to the last follow-up (P < 0.05). CONCLUSIONS: A 3D exoscope provides a visually detailed, deep and clear surgical field, which makes decompression safer and more effective and reduces short-term complications. A 3D exoscope may be a good assistance tool during decompression for single-segment massive LDH.

Integrin αVß3 Signaling in the Progression of Osteoarthritis Induced by Excessive Mechanical Stress.

Osteoarthritis (OA) is believed to be linked with cartilage degeneration, subchondral bone sclerosis, and synovial inflammation that lead to joint failure, and yet treatment that can effectively reverse the pathological process of the disease still not exists. Recent evidence suggests excessive mechanical stress (eMS) as an essential role in the pathogenesis of OA. Increased levels of integrin αVß3 have been detected in osteoarthritic cartilage and were previously implicated in OA pathogenesis. However, the role of integrin αVß3 in the process of eMS-induced OA remains unclear. Here, histologic and proteomic analyses of osteoarthritic cartilage in a rat destabilization of the medial meniscus model demonstrated elevated expression of integrin αVß3 as well as more serious cartilage degeneration in the medial weight-bearing area. Furthermore, results of in vitro study demonstrated that eMS led to a significant increase of integrin αVß3 expression and phosphorylation of downstream signaling molecules such as FAK and ERK, as well as upregulated expressions of inflammatory and degradative mediators. In addition, we found that inhibition of integrin αVß3 could alleviate chondrocyte inflammation triggered by eMS both in vivo and in vitro. Our findings suggest a central role for upregulation of integrin αVß3 signaling in OA pathogenesis and demonstrate that activation of integrin αVß3 signaling in cartilage contributes to inflammation and joint destruction in eMS-induced OA. Taken together, our data presented here provide a possibility for targeting integrin αVß3 signaling pathway as a disease-modifying therapy.

Size selection and placement of pedicle screws using robot-assisted versus fluoroscopy-guided techniques for thoracolumbar fractures: possible implications for the screw loosening rate.

BACKGROUND: There has been increased development of robotic technologies for the accuracy of percutaneous pedicle screw placement. However, it remains unclear whether the robot really optimize the selection of screw sizes and enhance screw stability. The purpose of this study is to compare the sizes (diameter and length), placement accuracy and the loosening rate of pedicle screws using robotic-assisted versus conventional fluoroscopy approaches for thoracolumbar fractures. METHODS: A retrospective cohort study was conducted to evaluate 70 consecutive patients [34 cases of robot-assisted percutaneous pedicle screw fixation (RAF) and 36 of conventional fluoroscopy-guided percutaneous pedicle screw fixation (FGF)]. Demographics, clinical characteristics, and radiological features were recorded. Pedicle screw length, diameter, and pedicle screw placement accuracy were assessed. The patients' sagittal kyphosis Cobb angles (KCA), anterior vertebral height ratios (VHA), and screw loosening rate were evaluated by radiographic data 1 year after surgery. RESULTS: There was no significant difference in the mean computed tomography (CT) Hounsfield unit (HU) values, operation duration, or length of hospital stay between the groups. Compared with the FGF group, the RAF group had a lower fluoroscopy frequency [14 (12-18) vs. 21 (16-25), P < 0.001] and a higher "grade A + B" pedicle screw placement rate (96.5% vs. 89.4%, P < 0.05). The mean screw diameter was 6.04 ± 0.55 mm in the RAF group and 5.78 ± 0.50 mm in the FGF group (P < 0.001). The mean screw length was 50.45 ± 4.37 mm in the RAF group and 48.63 ± 3.86 mm in the FGF group (P < 0.001). The correction loss of the KCA and VHR of the RAF group was less than that of the FGT group at the 1-year follow-up [(3.8 ± 1.8° vs. 4.9 ± 4.2°) and (5.5 ± 4.9% vs. 6.4 ± 5.7%)], and screw loosening occurred in 2 out of 34 patients (5.9%) in the RAF group, and 6 out of 36 patients (16.7%) in the FGF group, but there were no significant differences (P > 0.05). CONCLUSION: Compared with the fluoroscopy-guided technique, robotic-assisted spine surgery decreased radiation exposure and optimizes screw trajectories and dimensions intraoperatively. Although not statistically significant, the loosening rate of the RAF group was lower that of than the FGT group.

Nrf2 signaling activation by a small molecule activator compound 16 inhibits hydrogen peroxide-induced oxidative injury and death in osteoblasts.

We explored the potential activity of compound 16 (Cpd16), a novel small molecule Nrf2 activator, in hydrogen peroxide (H2O2)-stimulated osteoblasts. In the primary murine/human osteoblasts and MC3T3-E1 murine osteoblastic cells, Cpd16 treatment at micro-molar concentrations caused disassociation of Keap1-Nrf2 and Nrf2 cascade activation. Cpd16 induced stabilization of Nrf2 protein and its nuclear translocation, thereby increasing the antioxidant response elements (ARE) reporter activity and Nrf2 response genes transcription in murine and human osteoblasts. Significantly, Cpd16 mitigated oxidative injury in H2O2-stimulited osteoblasts. H2O2-provoked apoptosis as well as programmed necrosis in osteoblasts were significantly alleviated by the novel Nrf2 activator. Cpd16-induced Nrf2 activation and osteoblasts protection were stronger than other known Nrf2 activators. Dexamethasone- and nicotine-caused oxidative stress and death in osteoblasts were attenuated by Cpd16 as well. Cpd16-induced osteoblast cytoprotection was abolished by Nrf2 short hairpin RNA or knockout, but was mimicked by Keap1 knockout. Keap1 Cys151S mutation abolished Cpd16-induced Nrf2 cascade activation and osteoblasts protection against H2O2. Importantly, weekly Cpd16 administration largely ameliorated trabecular bone loss in ovariectomy mice. Together, Cpd16 alleviates H2O2-induced oxidative stress and death in osteoblasts by activating Nrf2 cascade.

Utility of a fulcrum for positioning support during flexion-extension radiographs for assessment of lumbar instability in patients with degenerative lumbar spondylolisthesis.

OBJECTIVE: The authors investigated a new standardized technique for evaluating lumbar stability in lumbar lateral flexion-extension (LFE) radiographs. For patients with lumbar spondylolisthesis, a three-part fulcrum with a support platform that included a semiarc leaning tool with armrests, a lifting platform for height adjustment, and a base for stability were used. Standard functional radiographs were used for comparison to determine whether adequate flexion-extension was acquired through use of the fulcrum method. METHODS: A total of 67 consecutive patients diagnosed with L4-5 degenerative lumbar spondylolisthesis were enrolled in the study. The authors analyzed LFE radiographs taken with the patient supported by a fulcrum (LFEF) and without a fulcrum. Sagittal translation (ST), segmental angulation (SA), posterior opening (PO), change in lumbar lordosis (CLL), and lumbar instability (LI) were measured for comparison using functional radiographs. RESULTS: The average value of SA was 5.76° ± 3.72° in LFE and 9.96° ± 4.00° in LFEF radiographs, with a significant difference between them (p < 0.05). ST and PO were also significantly greater in LFEF than in LFE. The detection rate of instability was 10.4% in LFE and 31.3% in LFEF, and the difference was significant. The CLL was 27.31° ± 11.96° in LFE and 37.07° ± 12.963.16° in LFEF, with a significant difference between these values (p < 0.05). CONCLUSIONS: Compared with traditional LFE radiographs, the LFEF radiographs significantly improved the detection rate of LI. In addition, this method may reduce patient discomfort during the process of obtaining radiographs.

A New Few-Shot Learning Method of Bacterial Colony Counting Based on the Edge Computing Device.

Bacterial colony counting is a time consuming but important task for many fields, such as food quality testing and pathogen detection, which own the high demand for accurate on-site testing. However, bacterial colonies are often overlapped, adherent with each other, and difficult to precisely process by traditional algorithms. The development of deep learning has brought new possibilities for bacterial colony counting, but deep learning networks usually require a large amount of training data and highly configured test equipment. The culture and annotation time of bacteria are costly, and professional deep learning workstations are too expensive and large to meet portable requirements. To solve these problems, we propose a lightweight improved YOLOv3 network based on the few-shot learning strategy, which is able to accomplish high detection accuracy with only five raw images and be deployed on a low-cost edge device. Compared with the traditional methods, our method improved the average accuracy from 64.3% to 97.4% and decreased the False Negative Rate from 32.1% to 1.5%. Our method could greatly improve the detection accuracy, realize the portability for on-site testing, and significantly save the cost of data collection and annotation over 80%, which brings more potential for bacterial colony counting.

Phosphoglycerate kinase 1 silencing by a novel microRNA microRNA-4523 protects human osteoblasts from dexamethasone through activation of Nrf2 signaling cascade.

Nuclear-factor-E2-related factor 2 (Nrf2) cascade activation can ameliorate dexamethasone (DEX)-induced oxidative injury and death in human osteoblasts. Phosphoglycerate kinase 1 (PGK1) depletion is shown to efficiently activate Nrf2 signaling by inducing methylglyoxal modification of Kelch-like ECH-associated protein 1 (Keap1). We here identified a novel PGK1-targeting microRNA: microRNA-4523 (miR-4523). RNA fluorescent in situ hybridization, RNA pull-down, and Argonaute-2 RNA immunoprecipitation results confirmed a direct binding between miR-4523 and PGK1 mRNA in primary human osteoblasts and hFOB1.19 osteoblastic cells. Forced overexpression of miR-4523, using a lentiviral construct, robustly decreased PGK1 3'-UTR (untranslated region) luciferase activity and downregulated its expression in human osteoblasts and hFOB1.19 cells. Furthermore, miR-4523 overexpression activated the Nrf2 signaling cascade, causing Keap1-Nrf2 disassociation, Nrf2 protein stabilization, and its nuclear translocation as well as transcription activation of Nrf2-dependent genes (NQO1, GCLC, and HO1) in human osteoblasts. By expressing a UTR-null PGK1 construct, miR-4523 overexpression-induced Nrf2 cascade activation was however largely inhibited. Importantly, DEX-induced reactive oxygen species production, oxidative injury, and cell apoptosis were significantly attenuated by miR-4523 overexpression in human osteoblasts and hFOB1.19 cells. Such actions by miR-4523 were abolished by Nrf2 shRNA or knockout, but mimicked by PGK1 knockout (using CRISPR/Cas9 method). In PGK1 knockout human osteoblasts, miR-4523 overexpression failed to further increase Nrf2 cascade activation and offer osteoblast cytoprotection against DEX. Significantly, miR-4523 is downregulated in human necrotic femoral head tissues of DEX-taking patients. Together, PGK1 silencing by miR-4523 protected human osteoblasts from DEX through activation of the Nrf2 signaling cascade.

[Study on Influencing Factors of the Tip Softness of Epidural Anesthesia Catheter].

This article aims to study the factors affecting the flexibility of the tip of an epidural anesthesia catheter. The flexibility of the tip of the epidural anesthesia catheter was tested with a softness tester from four aspects:raw materials, tip structure, tip processing technology, and the outer diameter of the catheter. Highly flexible and malleable polymer material with a smooth tip, the tip softening process and the proper outer diameter can effectively improve the tip flexibility of the epidural anesthesia catheter.

DCAF1-targeting microRNA-3175 activates Nrf2 signaling and inhibits dexamethasone-induced oxidative injury in human osteoblasts.

Activation of nuclear-factor-E2-related factor 2 (Nrf2) signaling can protect human osteoblasts from dexamethasone-induced oxidative injury. DDB1 and CUL4 associated factor 1 (DCAF1) is a novel ubiquitin E3 ligase for Nrf2 protein degradation. We identified a novel DCAF1-targeting miRNA, miR-3175. RNA pull-down, Argonaute 2 RNA-immunoprecipitation, and RNA fluorescent in situ hybridization results confirmed a direct binding between miR-3175 and DCAF1 mRNA in primary human osteoblasts. DCAF1 3'-untranslated region luciferase activity and its expression were significantly decreased after miR-3175 overexpression but were augmented with miR-3175 inhibition in human osteoblasts and hFOB1.19 osteoblastic cells. miR-3175 overexpression activated Nrf2 signaling, causing Nrf2 protein stabilization, antioxidant response (ARE) activity increase, and transcription activation of Nrf2-dependent genes in human osteoblasts and hFOB1.19 cells. Furthermore, dexamethasone-induced oxidative injury and apoptosis were largely attenuated by miR-3175 overexpression in human osteoblasts and hFOB1.19 cells. Importantly, shRNA-induced silencing or CRISPR/Cas9-mediated Nrf2 knockout abolished miR-3175 overexpression-induced osteoblast cytoprotection against dexamethasone. Conversely, DFAC1 knockout, by the CRISPR/Cas9 method, activated the Nrf2 cascade and inhibited dexamethasone-induced cytotoxicity in hFOB1.19 cells. Importantly, miR-3175 expression was decreased in necrotic femoral head tissues of dexamethasone-taking patients, where DCAF1 mRNA was upregulated. Together, silencing DCAF1 by miR-3175 activated Nrf2 signaling to inhibit dexamethasone-induced oxidative injury and apoptosis in human osteoblasts.

[Research progress of hair follicle and related stem cells in scar-free wound healing].

OBJECTIVE: To explore the research progress of hair follicle and related stem cells in scar-free skin healing in recent years. METHODS: The literature related to hair follicle and related stem cells, wound healing, and scar formation in recent years was extensively reviewed and summarized from the aspects of cell function and molecular mechanism. RESULTS: Scar tissue after wound healing treated with hair follicle transplantation and related stem cell therapy is more mild or even without scar formation. The cell types and molecular mechanisms of the above phenomena are complex, and the bone morphogenetic protein signal transduction pathway and Wnt signal transduction pathway are strongly correlated. CONCLUSION: The research of hair follicle and related stem cells in scar-free skin healing is at the initial stage at present. Strengthening the mechanism research may provide new ideas for the treatment of wound and scar.

Nitrogen vacancy mediated exciton dissociation in carbon nitride nanosheets: Enhanced hydroxyl radicals generation for efficient photocatalytic degradation of organic pollutants.

Polymeric materials are promising candidates as photocatalysts for environmental purification, however their catalytic performance are still unsatisfactory mainly due to the strong Coulomb interactions between electron and hole that leads to fast charge recombination. Herein, taking graphitic carbon nitride as an example, we verify that installing carbon nitride nanosheets with nitrogen vacancy could break the intrinsic electronic state distribution, forming energy disordered interfaces around the vacancies with the energy difference as large as 0.35 eV. Such a large energy difference is found energetic enough to overcome the strong Coulomb interactions between electron and hole for hot electron and hole generation, as a result showing high electron-hole separation efficiency. Benefited from these advantages, the as prepared material shows remarkable photocatalytic performance toward organic pollutants degradation. The improved catalytic performance is originated from the promoted exciton dissociation that leads to ultra high hydroxyl radical generation. This study offers a new understanding of the excitonic effects for designing advanced polymeric photocatalyst for energy and environment related applications.

Correlation Between Dural Sac Size in Dynamic Magnetic Resonance Imaging and Clinical Symptoms in Patients with Lumbar Spinal Stenosis.

OBJECTIVE: To assess the dynamic change of the dural sac size in patients with lumbar spinal stenosis (LSS) from supine to standing position and their correlation with clinical symptoms. METHODS: A total of 110 patients with LSS were prospectively enrolled to undergo both supine (0°) and standing (78°) magnetic resonance imaging (MRI). Dural sac cross-sectional area (DCSA) and dural sac anteroposterior diameter (DAPD) at the most constricted spinal level in supine and standing MRI were measured and compared. Clinical symptoms were assessed by duration of disease, claudication distance, visual analog scale (VAS) score of leg pain, and Chinese Oswestry Disability Index score of low back pain. The correlation between the parameters and clinical symptoms was analyzed by Pearson correlation coefficient (r). RESULTS: Mean minimum DCSA and DAPD in the standing position were significantly smaller (both P < 0.01) than in the supine position. DCSA and DAPD in standing MRI and their changes had better correlation with the intermittent claudication distance and VAS score of leg pain than in the supine position. A more than 15 mm2 reduction of DSCA was observed in patients with shorter claudication distance and more severe VAS score of leg pain (both P < 0.01). CONCLUSIONS: Dural sac size on MRI was reduced significantly from supine to standing position. Standing MRI and the changes of DCSA significantly correlated with claudication distance and VAS score of leg pain in patients with LSS. Therefore, standing MRI provides more radiologic information correlating with clinical symptoms in patients with LSS than supine MRI.