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Intervertebral disc degeneration (IVDD) severely affects the work and the quality of life of people. We previously demonstrated that silencing activation transcription factor 3 (ATF3) blocked the IVDD pathological process by regulating nucleus pulposus cell (NPC) ferroptosis, apoptosis, inflammation, and extracellular matrix (ECM) metabolism. Nevertheless, whether miR‐874‐3p mediated the IVDD pathological process by targeting ATF3 remains unclear. We performed single‐cell RNA sequencing (scRNA‐seq) and bioinformatics analysis to identify ATF3 as a key ferroptosis gene in IVDD. Then, Western blotting, flow cytometry, ELISA, and animal experiments were performed to validate the roles and regulatory mechanisms of miR‐874‐3p/ATF3 signalling axis in IVDD. ATF3 was highly expressed in IVDD patients and multiple cell types of IVDD rat, as revealed by scRNA‐seq and bioinformatics analysis. GO analysis unveiled the involvement of ATF3 in regulating cell apoptosis and ECM metabolism. Furthermore, we verified that miR‐874‐3p might protect against IVDD by inhibiting NPC ferroptosis, apoptosis, ECM degradation, and inflammatory response by targeting ATF3. In vivo experiments displayed the protective effect of miR‐874‐3p/ATF3 axis on IVDD. These findings propose the potential of miR‐874‐3p and ATF3 as biomarkers of IVDD and suggest that targeting the miR‐874‐3p/ATF3 axis may be a therapeutic target for IVDD. [ABSTRACT FROM AUTHOR]

Objective: Although neck pain has become a serious economic and social problem worldwide, the etiology remains poorly understood. The aim of current study is to explore the possible pathogenesis of discogenic neck pain by analyzing the relationship between inflammatory cytokines and discogenic neck pain and provide a valuable reference for the prevention and treatment of discogenic neck pain. Methods: A total of 111 cervical disc samples were collected between October 1, 2021, and October 1, 2022: 38 samples from the discogenic neck pain group, 41 samples from the symptomatic control group, and 32 samples from the normal control group. The concentration of nitric oxide (NO), interleukin (IL)‐1, interleukin (IL)‐6, and tumor necrosis factor alpha (TNF‐α) was determined using the enzyme‐linked immunosorbent assay in each sample, and the degeneration degree of the target discs were evaluated using T2‐weighted sagittal magnetic resonance imaging (MRI) according to the Miyazaki disc degeneration grading system. Whether the differences among the three groups were statistically significant was tested using one‐way analysis of variance and an unpaired t‐test, respectively. Results: The differences of the baseline characteristics were not statistically significant between the discogenic neck pain group and the symptomatic control group (p > 0.05). The expression of inflammatory cytokines in disc samples from the discogenic neck pain group (NO: 9.89 ± 1.75, IL‐1β: 10.74 ± 1.92, IL‐6:31.65 ± 2.46, and TNF‐α: 5.96 ± 1.91) was increased in comparison with the disc samples from both the symptomatic control group (NO: 7.15 ± 2.78, IL‐1β: 8.03 ± 1.87, IL‐6: 25.79 ± 2.12, and TNF‐α: 4.18 ± 2.87) and the normal control group (NO: 6.11 ± 1.37, IL‐1β: 5.84 ± 2.25, IL‐6: 20.65 ± 1.26, and TNF‐α: 2.05 ± 0.58). The differences were statistically significant (p < 0.001). Further, there were no statistical differences in the degree of degeneration between discogenic neck pain group and symptomatic control group. Conclusions: The increased expression of inflammatory cytokines in diseased cervical intervertebral discs might play a key role in the pathogenesis of discogenic neck pain. Although inflammation is involved in intervertebral disc degeneration, there is no linear positive correlation between the concentration of inflammatory cytokines and the degree of disc degeneration. [ABSTRACT FROM AUTHOR]

Purpose: To explore the predictive factors of neck pain (NP) in patients with cervical degenerative disease by retrospectively analyzing their occupational and demographic characteristics and to provide a valuable reference for preventing and treating chronic NP.Patients and Methods: We retrospectively reviewed the occupational and demographic data of patients with cervical degenerative disease who had undergone anterior cervical surgery between June 2021 and December 2022 at our center. The patients were divided into NP and no-NP groups based on whether they had chronic NP before surgery. Relevant occupational and demographic data from all patients were statistically analyzed, and all variables were made categorical. Forward stepwise logistic regression models were constructed for preoperative chronic neck pain to explore the possible risk factors associated with chronic neck pain.Results: The differences in smoking, being an office worker, BMI, and disease types between NP and no-NP groups were statistically significant. In contrast, there were no statistically significant in age, sex, academic level, duration, and degeneration grade between the two groups. Moreover, further logistic regression analysis indicated that smoking, being an office worker, having an abnormal BMI, and cervical spondylotic radiculopathy (CSR) were related to chronic neck pain.Conclusion: The present study indicated that smoking, being an office worker, having an abnormal BMI, and CSR were predisposing risk factors for NP associated with cervical degenerative disease. Although intervertebral disc degeneration is the pathology basis of NP, the degeneration grade was not related to the occurrence of NP in our current study. Therefore, quitting smoking, avoiding sedentariness, and maintaining a normal BMI may prevent NP to some extent. [ABSTRACT FROM AUTHOR]

Study Design: This is a retrospective study. Objective: The aim of the study was to evaluate the efficacy of self-anchored lateral lumbar interbody fusion (SA-LLIF) in lumbar degenerative diseases. Methods: Forty-eight patients with lumbar degenerative disease between January 2019 and June 2020 were enrolled in this study. All patients complained of low back and leg pain, which were aggravated during standing activities and alleviated or disappeared during lying. After general anesthesia, the patient was placed in the right decubitus position. The anterior edge of the psoas major muscle was exposed through an oblique incision of approximately 6 cm, using an extraperitoneal approach. The psoas major muscle was then properly retracted dorsally to expose the disc. After discectomy, a suitable cage filled with autogenous bone graft from the ilium was implanted. Two anchoring plates were inserted separately into the caudal and cranial vertebral bodies to lock the cage. Clinical efficacy was evaluated using the visual analog scale (VAS) and Oswestry Disability Index (ODI). Lumbar lordosis, intervertebral disc height, spondylolisthesis rate, cage subsidence and fusion rate were also recorded. Results: A total of 48 patients were enrolled in this study, including 20 males and 28 females, aged 61.4 ± 7.3 (range 49–78) years old. Surgery was successfully performed in all patients. Lumbar stenosis and instability were observed in 22 cases, disc degenerative disease in eight cases, degenerative spondylolisthesis in nine cases, degenerative scoliosis in six cases, and postoperative revision in three cases. In addition, five patients were diagnosed with osteoporosis. The index levels included L2–3 in three patients, L3–4 in 13 patients, L4–5 in 23 patients, L2–4 in three patients, and L3–5 in six patients. The operation time was 81.1 ± 6.4 (range 65–102) min. Intraoperative blood loss was 39.9 ± 8.5 (range 15–72) mL. No severe complications occurred, such as nerve or blood vessel injuries. The patients were followed up for 11.7 ± 2.3 (range 4–18) months. At the last follow-up, the VAS decreased from 6.2 ± 2.3 to 1.7 ± 1.1, and the ODI decreased from 48.4% ± 11.2% to 10.9% ± 5.5%. Radiography showed satisfactory postoperative spine alignment. No cage displacement was found, but cage subsidence 2–3 mm was found in five patients without obvious symptoms, except transient low back pain in an obese patient. The lumbar lordosis recovered from 36.8° ± 7.9° to 47.7° ± 6.8°, and intervertebral disc height recovered from 8.2 ± 2.0 mm to 11.4 ± 2.5 mm. The spondylolisthesis rate decreased from 19.9% ± 4.9% to 9.4% ± 3.2%. The difference between preoperative and last follow-up was statistically significant (P<0.05). Conclusion: SA-LLIF can provide immediate stability and good results for lumbar degenerative diseases with a standalone anchored cage without posterior internal fixation. [ABSTRACT FROM AUTHOR]

Background. The competing endogenous RNA- (ceRNA-) mediated regulatory mechanisms are known to play a pivotal role in intervertebral disc degeneration (IDD). Our research intended to establish a ceRNA regulatory network related to IDD through bioinformatics analyses. Methods. The expression profiles of circRNA, miRNA, and mRNA were obtained from the public Gene Expression Omnibus (GEO) datasets. Then, we use sequence-based bioinformatics methods to select differentially expressed mRNAs (DEmRNAs), microRNAs (DEmiRNAs), or circRNAs (DEcircRNAs) related to IDD. We used ChEA3 to verify the targets of transcription factors (TFs). Then, we used DAVID to annotate the DEmRNAs. Finally, we constructed a potentially circRNA-miRNA-mRNA network related to IDD by predicting in the database (ENCORI, TargetScan, miRecords, miRmap, and circBank). Results. We identified 31 common DEmRNAs by Venn analysis, of which MMP2 was regarded as the key hub genes. Simultaneously, miR-423-5p and miR-185-5p were predicted as the upstream molecules of MMP2. Furthermore, a total of six DEcircRNAs were predicted as the upstream circRNAs of miR-423-5p and miR-185-5p. Then, a potential circRNA-miRNA-mRNA network related to IDD was constructed by bioinformatics analysis. Conclusion. A comprehensive ceRNA regulatory network was constructed, which was found to be significant in IDD progression. [ABSTRACT FROM AUTHOR]

Objective. Discectomy remains the classic procedure for treating lumbar intervertebral disc (IVD) herniation, but the occurrence of defects after discectomy is thought to be an important cause generating recurrent and accelerated IVD degeneration. Previous studies attempted suture of the annulus fissure, but the validity of this technique on restraining the degenerative process is controversial. On the other hand, cell therapies have been shown in multiple clinical and basic studies. Our purpose was to investigate the effectiveness of selective retention of autologous Bone Marrow Stromal Cells (BMSCs) with gelatin sponge in combination with annulus fibrosus suture (AFS) for the repair of IVD defects following mobile microendoscopic discectomy (MMED). Methods. This prospective, two-armed, and controlled clinical study was conducted from December 2016 to December 2018. Written informed consent was obtained from each patient. Forty-five patients with typical symptoms, positive signs of radiculopathy, and obvious lumbar disc herniation observed by MRI were enrolled. Patients were divided into 3 groups with different treating methods: MMED (n = 15), MMED+AFS (n = 15), and MMED+AFS+BMSCs (n = 15). A postoperative 2-year follow-up was performed to evaluate the patient-reported outcomes of VAS, ODI, and SF-36. The improvement rate of VAS and ODI was calculated as latest ‐ preoperative / preoperative to evaluate the therapeutic effect of the three groups. Assessment parameters included Pfirrmann grade, intervertebral disc height (IDH), and disc protrusion size (DPS), as measured by MRI to evaluate the morphological changes. Results. All patients enrolled had a postoperative follow-up at 3, 6, 12, and 24 months. VAS and ODI scores were significantly improved compared to the preoperative status in all three groups with a mean DPS reduction rate over 50%. At the final follow-up, the improvement rate of the VAS score in the MMED+AFS+BMSCs group was significantly higher than the MMED+AFS and MMED groups (80.1 % ± 7.6 % vs. 71.3 % ± 7.0 % vs. 70.1 % ± 7.8 %), while ODI improvement showed a significant change (65.6 % ± 8.8 % vs. 59.9 % ± 5.5 % vs. 57.8 % ± 8.1 %). All participants showed significant improvement in SF-36 PCS and MCS; the differences between each group were not significant. The mean IDH loss rate of the MMED+AFS+BMSCs group was also significantly lower than other groups (− 17.2 % ± 1.3 % vs. − 27.6 % ± 0.7 % vs. − 29.3 % ± 2.2 %). The Pfirrmann grade was aggravated in the MMED and MMED+AFS groups while maintained at the preoperative grade in the MMED+AFS+BMSCs group. No adverse events of cell transplantation or recurrence were found in all patients during the postoperative follow-up period. Conclusions. It is feasible and effective to repair lumbar IVD defects using SCR-enriched BMSCs with gelatin sponges, which warrants further study and development as a cell-based therapy for IVD repair. [ABSTRACT FROM AUTHOR]

The functional alteration of nucleus pulposus cells (NPCs) exerts a crucial role in the occurrence and progression of intervertebral disk degeneration (IDD). Circular RNAs and microRNAs (miRs) are critical regulators of NPC metabolic processes such as growth and apoptosis. In this study, bioinformatics tools, encompassing Gene Ontology pathway and Venn diagrams analysis, and protein–protein interaction (PPI) network construction were used to identify functional molecules related to IDD. PPI network unveiled that ESR1 was one of the most critical genes in IDD. Then, a key IDD-related circ_0040039-miR-874-3p-ESR1 interaction network was predicted and constructed. Circ_0040039 promoted miR-874-3p and repressed ESR1 expression, and miR-874-3p repressed ESR1 expression in NPCs, suggesting ESR1 might be a direct target of miR-874-3p. Functionally, circ_0040039 could enhance NPC apoptosis and inhibit NPC growth, revealing that circ_0040039 might aggravate IDD by stabilizing miR-874-3p and further upregulating the miR-874-3p-ESR1 pathway. This signaling pathway might provide a novel therapeutic strategy and targets for the diagnosis and therapy of IDD-related diseases. [ABSTRACT FROM AUTHOR]

Background. Intervertebral disc degeneration (IDD) disease is a global challenge because of its predominant pathogenic factor in triggering low back pain, whereas cartilaginous endplate degeneration (CEPD) is the main cause of IDD. Accumulating evidence have indicated that the differentially expressed microRNAs (DEMs) and differentially expressed genes (DEGs) have been determined to be involved in multiple biological processes to mediate CEPD progression. However, the differentially expressed circular RNAs (DECs) and their potential biofunctions in CEPD have not been identified. Methods. GSE153761 dataset was analyzed using R software to predict DECs, DEMs, and DEGs. Pathway enrichment analysis of DEGs and host genes of DECs and protein-protein interaction network of DEGs were conducted to explore their potential biofunctions. Furthermore, we explore the potential relationship between DEGs and DECs. Results. There were 74 DECs, 17 DEMs, and 68 DEGs upregulated whereas 50 DECs, 16 DEMs, and 67 DEGs downregulated in CEPD group. Pathway analysis unveiled that these RNAs might regulate CEPD via mediating inflammatory response, ECM metabolism, chondrocytes apoptosis, and chondrocytes growth. A total of 17 overlapping genes were predicted between the host genes of DEGs and DECs, such as SDC1 and MAOA. Moreover, 6 upregulated DECs, of which hsa_circ_0052830 was the most upregulated circRNA in CEPD, were derived from the host genes SDC1, whereas 8 downregulated DECs were derived from the host genes MAOA. Conclusion. This will provide novel clues for future experimental studies to elucidate the pathomechanism of CEPD and therapeutic targets for CEPD-related diseases. [ABSTRACT FROM AUTHOR]

Tissue engineering has provided an alternative therapeutic possibility for degenerative disc diseases. However, we lack an ideal scaffold for IVD tissue engineering. The goal of this study is to fabricate a novel biomimetic biphasic scaffold for IVD tissue engineering and evaluate the feasibility of developing tissue-engineered IVD in vitro and in vivo. In present study we developed a novel integrated biphasic IVD scaffold using a simple freeze-drying and cross-linking technique of pig bone matrix gelatin (BMG) for the outer annulus fibrosus (AF) phase and pig acellular cartilage ECM (ACECM) for the inner nucleus pulposus (NP) phase. Histology and SEM results indicated no residual cells remaining in the scaffold that featured an interconnected porous microstructure (pore size of AF and NP phase 401.4±13.1 μm and 231.6±57.2 μm, respectively). PKH26-labeled AF and NP cells were seeded into the scaffold and cultured in vitro. SEM confirmed that seeded cells could anchor onto the scaffold. Live/dead staining showed that live cells (green fluorescence) were distributed in the scaffold, with no dead cells (red fluorescence) being found. The cell—scaffold constructs were implanted subcutaneously into nude mice and cultured for 6 weeks in vivo. IVD-like tissue formed in nude mice as confirmed by histology. Cells in hybrid constructs originated from PKH26-labeled cells, as confirmed by in vivo fluorescence imaging system. In conclusion, the study demonstrates the feasibility of developing a tissue-engineered IVD in vivo with a BMG- and ACECM-derived integrated AF-NP biphasic scaffold. As well, PKH26 fluorescent labeling with in vivo fluorescent imaging can be used to track cells and analyse cell—scaffold constructs in vivo. [ABSTRACT FROM AUTHOR]

Objective To investigate the application of PKH26 fluorescent labeling on nucleus pulposus cells isolated from bovine coccyx disc, and to provide nucleus pulposus tissue engineering with traceable nucleus pulposus cells by PKH26 fluorescence labelling. Methods Nucleus pulposus primary cells were isolated from the nucleus pulposus tissue detached from bovine coccyx disc by enzymatic digestion, and observed under the inverted microscope. Safranin O, toluidine blue and type II collagen immunocytochemistry methods used to stain for passage one generation cells. Nucleus pulposus cells were labeled with PKH26 fluorescence in accordance with the instructions. The cell activity, fluorescence intensity at d0, d14 and d28 of culture, characteristics of proliferation and the expression of gene in labeled cells were assessed. Results Isolated nucleus pulposus cells amounted to (1.56 ± 0.35) x 106/g. Under the inverted microscope, primary cells adhered at the 4 th day of culture, grew in groups, and covered the bottom of culture flask at the 13 th day. Both primary cells and the P1 generation cells were chondrocyte-like morphology. The staining of safranin O, toluidine blue and type II collagen immunocytochemistry for P1 generation of nucleus pulposus cells showed positive results. The cell activity before and after PKH26 labeling showed more than 95%, and the fluorescence intensity at d0, d14 and d28 performed a decreasing trend, but still showed detect strong fluorescence at d28. There were no significant differences in proliferation and the expression of gene (collagen type I and II, aggrecan) before and after cell labeling (P > 0.05). Conclusion As the seed cells of tissue engineering, nucleus pulposus cells isolated from bovine coccyx can reach a satisfactory number and maintain cartilage-like phenotype, and no changes shown in the biological characteristics after labeling. PKH26 labeled nucleus pulposus cells are suitable for the traceable cells in vivo study. [ABSTRACT FROM AUTHOR]

Abstract: The tissue-engineered intervertebral disc (IVD) construct offers an alternative therapeutic possibility for symptomatic degenerative disc diseases. A porcine-derived acellular matrix as an integrated biphasic IVD scaffold was fabricated using an optimal decellularization procedure with a simple freeze-drying and cross-linking technique. The biphasic construct contained annulus fibrosus (AF) phase with cancellous bone matrix gelatin (CBMG) and the nucleus pulposus (NP) phase with cartilage extracellular matrix (CECM). In addition, this decellularized IVD scaffold possessed interconnected and tightly integrated porous microstructure (pore size of AF and NP phase at 401.4±13.1μm and 231.6±57.2μm, respectively), and had appropriate mechanical property. Histological results showed no residual cells remaining within the decellularized scaffold but abundant collagen proteins retained. Fluorescence microscope and scanning electron microscopy demonstrated that the novel scaffold could provide a suitable 3-D environment to support adhesion and proliferation of both AF and NP cells. In conclusion, this xenogenous ECM-derived IVD scaffold provides potential as an alternative for IVD tissue engineering. [Copyright &y& Elsevier]