JOR Spine最新文献

{"title":"Application and Validation of Semiautomatic Quantification of Immunohistochemically Stained Sections for Low Cellular Tissue Such as Intervertebral Disc Using QuPath","authors":"Andrea Nüesch,&nbsp;Maria Paola Ferri,&nbsp;Christine L. Le Maitre","doi":"10.1002/jsp2.70054","DOIUrl":"https://doi.org/10.1002/jsp2.70054","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Immunohistochemistry (IHC) is a widely used method for localizing and semi-quantifying proteins in tissue samples. Traditional IHC analysis often relies on manually counting 200 cells within a designated area, a time-intensive and subjective process that can compromise reproducibility and accuracy. Advances in digital scanning and bioimage analysis tools, such as the open-source software QuPath, enable semi-automated cell counting, reducing subjectivity and increasing efficiency.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Aims</h3>\u0000 \u0000 <p>This project developed a QuPath-based script and detailed guide for semi-automatic cell counting, specifically for tissues with low cellularity, such as intervertebral discs and cartilage.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods and Results</h3>\u0000 \u0000 <p>The methodology was validated by demonstrating no significant differences between the manual counting and the semi-automatic quantification (<i>p</i> = 0.783, <i>p</i> = 0.386) while showing a strong correlation between methods for both collagen type II staining (<i>r</i> = 0.9602, <i>p</i> &lt; 0.0001) and N-cadherin staining (<i>r</i> = 0.9044, <i>p</i> = 0.0001). Furthermore, a strong correlation (intraclass correlation coefficient (ICC) single raters = 0.853) between 3 individual raters with varying academic ranks and experiences in IHC analysis was shown using the semi-automatic quantification method.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Discusssion</h3>\u0000 \u0000 <p>The approach ensures high reproducibility and accuracy, with reduced variability between raters and laboratories. This semi-automated method is particularly suited for tissues with a high extracellular matrix to cell ratio and low cellularity. By minimizing subjectivity and evaluation time, it provides a robust alternative to manual counting, making it ideal for applications where reproducibility and standardization are critical. While the methodology was effective in low-cellularity tissues, its application in other tissue types warrants further exploration.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>These findings underscore the potential of QuPath to streamline IHC analysis and enhance inter-laboratory comparability.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143565143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Significance of Cellular Senescence Hub Genes in the Diagnosis and Subtype Classification of a Comprehensive Database of Gene Expression in Intervertebral Disc Degeneration","authors":"Fei Liu,&nbsp;Silong Gao,&nbsp;Ji Yin,&nbsp;Chao Song,&nbsp;Yongliang Mei,&nbsp;Zhaoqiang Wang,&nbsp;Zongchao Liu","doi":"10.1002/jsp2.70050","DOIUrl":"https://doi.org/10.1002/jsp2.70050","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Intervertebral disc degeneration (IVDD) is a complex age-related physiological process, with cellular senescence (CS) being a primary contributing factor. However, the precise role of CS and its associated genes in IVDD remains unclear.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In this study, we performed differential expression analysis on the GSE124272 and GSE150408 datasets from the GEO database and identified 53 differentially expressed cellular senescence-related genes (CSRGs). We then conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to explore their functions and associated pathways. We identified hub genes by constructing a protein–protein interaction (PPI) network and further validated these genes using clinical samples. We further explored the functional and prognostic significance of these genes using support vector machine recursive feature elimination (SVM-RFE), random forest (RF), and least absolute shrinkage and selection operator (LASSO) algorithms. We visualized the correlation between the differential expression levels of the four core genes and immune cell infiltration using heat maps and histograms. Finally, we performed graphene oxide enrichment analysis on 297 differentially expressed genes (DEGs) to investigate their role in IVDD.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>We ultimately identified four hub cellular CSRGs DUSP3, MAPKAPK5, SP1, and VEGFA, and further validated their expression using various algorithms and clinical samples. Our results revealed that DUSP3 and SP1 were upregulated in IVDD, while MAPKAPK5 and VEGFA were downregulated. Immune cell infiltration analysis demonstrated that DUSP3 and SP1 were positively correlated with immune cell infiltration levels, whereas VEGFA and MAPKAPK5 were negatively correlated.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>In summary, CSRGs play an important role in the pathogenesis of IVDD, and our study of the hub gene cluster may guide future therapeutic strategies for IVDD.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70050","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143565144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DeVa (Decay Variance): A Novel Score Calculated via Postprocessing the Changes in Signal Intensity of an Intervertebral Disc in a T2* Multi-Echo Magnetic Resonance Image Can Quantify Painful and Degenerate Lumbar Vertebral Discs","authors":"Stone Sima,&nbsp;Alisha Sial,&nbsp;Suhani Sharma,&nbsp;Dheera Ananthakrishnan,&nbsp;Jeff Kuan,&nbsp;Ashish Diwan","doi":"10.1002/jsp2.70056","DOIUrl":"https://doi.org/10.1002/jsp2.70056","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Introduction</h3>\u0000 \u0000 <p>Low back pain (LBP), a global disability leader, is often linked to intervertebral disc (IVD) degeneration. Traditional diagnostics like T2-weighted MRI provide qualitative but imprecise evaluations. A novel post-processing MRI technique, Decay Variance (DeVa), has shown promise in differentiating degenerate from healthy discs in animal studies. DeVa quantifies IVD degeneration by analyzing variations in signal intensities within each voxel in a T2* 2D FLASH multi-echo MRI sequence. This study aimed to validate DeVa clinically and explore its correlation with pain severity.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>A cross-sectional study included 77 chronic LBP patients and 8 controls, who underwent T2-weighted and T2* 2D FLASH MRI. DeVa scores (worst and sum of all discs) were recorded, alongside traditional assessments like disc bulge, stenosis, high-intensity zones, and Pfirrmann grade. Pain severity was measured with a numerical rating scale. Statistical analyses included Pearson correlation, <i>t</i>-tests, and Gardner-Altman plots to evaluate relationships between DeVa scores, degeneration, and pain.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>DeVa scores correlated strongly with Pfirrmann grade (<i>r</i> = 0.692, <i>p</i> &lt; 0.001) and were significantly higher in discs with bulge, stenosis, or high-intensity zones (<i>p</i> &lt; 0.001). Moderate correlations were observed between worst DeVa scores (<i>r</i> = 0.296, <i>p</i> &lt; 0.01), total DeVa scores (<i>r</i> = 0.323, <i>p</i> &lt; 0.005) and pain severity. Patients with chronic LBP without severe degeneration (Pfirrmann ≤ 3 with no stenosis observable on standard MRI) had significantly higher worst (1.38 ± 0.26 vs. 1.10 ± 0.29, <i>p</i> &lt; 0.005) and total (5.39 ± 0.75 vs. 4.65 ± 0.61, <i>p</i> &lt; 0.0.1) DeVa scores compared to controls.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Discussion</h3>\u0000 \u0000 <p>DeVa offers a quantitative, noninvasive approach to assessing IVD degeneration, showing strong correlations with disc health and pain. It demonstrates enhanced sensitivity over traditional MRI, enabling the identification of pain-generating discs and informing personalized treatment strategies for chronic LBP. Further validation in larger populations is needed.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70056","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143565145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ruxolitinib Delays Nucleus Pulposus Cell Senescence in Rat Intervertebral Discs","authors":"Honggang Hao,&nbsp;Weidong Liang,&nbsp;Shuwen Zhang,&nbsp;Xiaoyu Cai,&nbsp;Abulizi Yakefu,&nbsp;Shutao Gao,&nbsp;Chuanhui Xun,&nbsp;Tao Xu,&nbsp;Rui Cao,&nbsp;Weibin Sheng","doi":"10.1002/jsp2.70044","DOIUrl":"https://doi.org/10.1002/jsp2.70044","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Intervertebral disc degeneration involves aging and senescence of nucleus pulposus cells (NPCs), and JAK/STAT signaling may contribute to this process. The aim of this study was to investigate the therapeutic effect of the JAK2 inhibitor, ruxolitinib, on NPC senescence.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Control (third passage), Senescence (sixth passage), JAK inhibitor (ruxolitinib-treated), siRNA-NC (control siRNA-treated), and siRNA-JAK2 (JAK2-targeting siRNA-treated) groups of rat NPCs were established. Cell senescence ratios were determined by β-galactosidase staining and Edu staining was conducted to assess cell proliferation. Cell cycle and apoptosis were analyzed by flow cytometry and Aggrecan and Col II expression detected by immunofluorescence staining. Levels of IL-1β, IL-6, TNF-α, MMP-3, and MMP-13 were detected by ELISA, and p16, p21, p53, p-p53, JAK2, STAT3, p-JAK2, p-STAT3, ADAMTS4, and ADAMTS5 levels were examined by western blot.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>More cell senescence was detected by β-galactosidase staining in the Senescence group than in the Control group, while cell proliferation was lower, apoptosis ratio higher, and the percentage of NPCs in G0/G1 phase higher. Levels of senescence-related proteins, including p16, p21, p53, and p-p53, were higher in the Senescence group than the Control group, as were those of IL-1β, IL-6, TNF-α, MMP-3, MMP-13, ADAMTS4, and ADAMTS5. Further, Aggrecan and Col II levels were lower in the Senescence group, while those of JAK2 and STAT3 (JAK2/STAT3 signaling pathway) were higher. Ruxolitinib reversed the changes described above to varying degrees, and the results were supported by those of experiments involving targeted silencing of JAK2.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>NPC senescence is characterized by low cell proliferation, a high apoptosis ratio, cell cycle arrest, and generation of senescence-associated secretory phenotypes. NPC senescence can be delayed by inhibiting JAK2/STAT3 signaling using ruxolitinib.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70044","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143554538","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Range of Motion and Neutral Zone of All Human Spinal Motion Segments: A Data Collection of 30 Years of In Vitro Experiments Performed Under Standardized Testing Conditions","authors":"Hans-Joachim Wilke,&nbsp;Annette Kienle,&nbsp;Karin Werner,&nbsp;Christian Liebsch","doi":"10.1002/jsp2.70052","DOIUrl":"https://doi.org/10.1002/jsp2.70052","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Spinal flexibility can vary among spinal sections and single motion segments. The purpose of this work was to provide a comprehensive overview of the range of motion (RoM) and neutral zone (NZ) values of all spinal levels collected during 30 years of in vitro experiments under standardized testing conditions.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>RoM and NZ data obtained from in vitro testing of intact human mono- and polysegmental specimens with pure moments of 2.5 Nm for the cervical, 5 Nm for the thoracic, and 7.5 Nm for the lumbar spine were collated from the internal database of the authors' institution. Descriptive statistics were performed with median values and median absolute deviations. Outliers were defined as values beyond twofold standard deviation and excluded from evaluation. Normal distribution was verified using the Shapiro–Wilk test.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>RoM and NZ data of <i>N</i> = 1139 functional spinal units were collected with sample sizes ranging from <i>n</i> = 6 to <i>n</i> = 224 per segmental level. The cervical spine was very flexible in flexion/extension and moderately flexible in lateral bending, while in axial rotation, the motion segment C1-C2 was as flexible as the subaxial cervical spine combined. The thoracic spine was the least flexible section in flexion/extension but allowed moderate lateral bending and axial rotation. The rib cage had a strong effect on thoracic spinal flexibility, particularly in axial rotation and at the mid-thoracic spinal levels. The lumbar spine exhibited moderate flexibility in flexion/extension and lateral bending but showed the lowest RoM and NZ in axial rotation.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This unique summary of RoM and NZ data, acquired under the same loading conditions in the same spine tester, provides a detailed insight into overall spinal flexibility and will serve as a valid dataset for the validation of in vitro studies and numerical models of the single motion segments of the spine.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143554535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanism of Intervertebral Disc Degeneration via the β-Catenin/CCL2 Pathway in Sox9 Conditional Knockout Mice","authors":"Khaled Aboushaala,&nbsp;Ana Chee,&nbsp;Frank Ko,&nbsp;Jad Alkhudari,&nbsp;Saurav Sumughan,&nbsp;Howard S. An,&nbsp;Dino Samartzis,&nbsp;Chun-do Oh","doi":"10.1002/jsp2.70053","DOIUrl":"https://doi.org/10.1002/jsp2.70053","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Introduction</h3>\u0000 \u0000 <p>Degenerative changes in the intervertebral disc (IVD) are known to be a main cause of low back pain (LBP), oftentimes necessitating interventions that may or may not be successful due to a lack of understanding in the degenerative phenotype and its mechanisms. Understanding the molecular mechanisms of disc degeneration can help design new therapies to induce disc regeneration and reduce back pain. This work aimed to understand the effects of conditional deletion of Sox9 in aggrecan-expressing cells on intervertebral disc degeneration and its underlying mechanisms in mice.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>This study utilized <i>Agc1-CreERT2;Sox9</i>^{<i>flox/flox</i>} mice to investigate the effects of SOX9 deletion on IVD degeneration and associated pain behaviors. Mice were administered tamoxifen to induce conditional gene deletion of Sox9. Structural and degenerative phenotypes of the spine were assessed by a histological scoring system and micro-computed tomography (microCT). Pain behaviors were evaluated through mechanical allodynia testing and the LABORAS system for spontaneous behavior assessment. Immunohistochemistry identified the expression of proteins of interest, which were further examined by Western blotting. Lastly, quantitative real-time PCR and promoter assays on IVD cells were used to examine inflammatory and signaling pathways induced by Sox9 deletion.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Crossing <i>Agc1-CreERT2</i> mice with <i>Sox9</i>^{<i>flox/flox</i>} mice revealed that <i>Sox9</i> conditional deletion (<i>Sox9</i>^<i>cKO</i>) in cartilage tissues causes IVD degeneration and pain behavior. <i>Sox9</i>^<i>cKO</i> mice spines had narrowed intervertebral disc spaces and disorganized IVD tissues. <i>Sox9</i> deletion also increased β-catenin, C-C motif chemokine ligand 2 (CCL2), and Glial cell line-derived neurotrophic factor (GDNF) expression in the IVD, suggesting their roles in disc pain and degeneration and the importance of the β-catenin/CCL2 pathway in these processes.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Deletion of <i>Sox9</i> in Aggrecan-expressing IVD tissues affects disc degeneration and associated pain behaviors through the β–catenin–CCL2 pathway. Such findings can lead to more targeted, personalized therapeutics in the future to address discogenic origins of LBP.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70053","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143497306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gene Expression Changes Precede Elevated Mechanical Sensitivity in the Mouse Intervertebral Disc Injury Model","authors":"Zuozhen Tian,&nbsp;Ken Chen,&nbsp;Frances S. Shofer,&nbsp;Brianna Ciesielski,&nbsp;Huan Wang,&nbsp;W. Timothy O'Brien,&nbsp;Ling Qin,&nbsp;Yejia Zhang","doi":"10.1002/jsp2.70049","DOIUrl":"https://doi.org/10.1002/jsp2.70049","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Back pain after intervertebral disc (IVD) injury is a common clinical problem. Previous work examining early molecular changes post injury mainly used a candidate marker approach.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In this study, gene expression in the injured and intact mouse tail IVDs was determined with a nonbiased whole transcriptome approach and related to subsequent pain behavior. Mouse tail IVD injury was induced by a needle puncture. Whole murine transcriptome was determined by RNASeq. Transcriptomes of injured IVDs were compared with those of intact controls by bioinformatic methods. Mechanical allodynia was assessed by the Von Frey method.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Among the 17,722 murine genes with meaningful expressions, 7242 genes were differentially expressed (<i>P.adj</i> ⟨ 0.01). Ontology study of upregulated genes revealed that leukocyte migration was the most enriched biological process, and network analysis showed that Tnfa had the most protein–protein interactions. The most enriched downregulated pathways were related to the pattern specification process. Mechanical allodynia persisted at the 4-week end point.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The RNASeq data revealed numerous early genes that participate in inflammation and repair processes post IVD injury. Mechanical allodynia followed these gene expression changes.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143475409","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Parallel Networks to Predict TIMP and Protease Cell Activity of Nucleus Pulposus Cells Exposed and Not Exposed to Pro-Inflammatory Cytokines","authors":"L. Baumgartner,&nbsp;S. Witta,&nbsp;J. Noailly","doi":"10.1002/jsp2.70051","DOIUrl":"https://doi.org/10.1002/jsp2.70051","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Intervertebral disc (IVD) degeneration is characterized by a disruption of the balance between anabolic and catabolic cellular processes. Within the nucleus pulposus (NP), this involves increased levels of the pro-inflammatory cytokines interleukin 1beta (IL1B) and tumor necrosis factor (TNF) and an upregulation of the protease families matrix metalloproteinase (MMP) and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS). Primary inhibitors of these proteases are the tissue inhibitors of matrix metalloproteinases (TIMP). This work aims at contributing to a better understanding of the dynamics among proteases, TIMP, and pro-inflammatory cytokines within the complex, multifactorial environment of the NP.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>The Parallel Network (PN)-Methodology was used to estimate relative mRNA expressions of TIMP1–3, MMP3, and ADAMTS4 for five simulated human activities: walking, sitting, jogging, hiking with 20 kg extra weight, and exposure to high vibration. Simulations were executed for nutrient conditions in non- and early-degenerated IVD approximations. To estimate the impact of cytokines, the PN-Methodology inferred relative protein levels for IL1B and TNF, reintegrated as secondary stimuli into the network.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>TIMP1 and TIMP2 expressions were found to be overall lower than TIMP3 expression. In the absence of pro-inflammatory cytokines, MMP3 and/or ADAMTS4 expressions were strongly downregulated in all conditions but vibration and hiking with extra weight. Pro-inflammatory cytokine exposure resulted in an impaired inhibition of MMP3, rather than of ADAMTS4, progressively rising with increasing nutrient deprivation. TNF mRNA was less expressed than IL1B. However, at the protein level, TNF was mainly responsible for the catabolic shift in the simulated pro-inflammatory environment. Overall, results agreed with previous experimental findings.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The PN-Methodology successfully allowed the exploration of the relative dynamics of TIMP and protease regulations in different mechanical, nutritional, and inflammatory environments in the NP. It shall stand as a comprehensive tool to integrate in vitro model results in IVD research and approximate NP cell activities in complex multifactorial environments.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70051","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143447211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Do we know more about the mechanobiology of the intervertebral disc in space than on Earth?","authors":"Timothy Patrick Holsgrove,&nbsp;Isabelle Ebisch,&nbsp;Daniela Lazaro-Pacheco","doi":"10.1002/jsp2.70024","DOIUrl":"https://doi.org/10.1002/jsp2.70024","url":null,"abstract":"<p>This work provides a perspective on the loading protocols used in whole-organ interverterbal disc culture studies using bioreactors. We put this in the context of in vivo spinal loading, and we put forward the case that the majority of previous bioreactor studies have more in common with spinal loading in space than on Earth. Finally, we provide an outlook for the future of bioreactor research, to provide data more relevant to spinal loading on Earth, and maximize the translational potential of findings to the clinical setting.\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure></p>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143431737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transient Receptor Potential Vanilloid 4 Knockdown Decreases Extracellular Matrix Synthesis via Autophagy Suppression in the Rat Intervertebral Disc","authors":"Tomoya Matsuo,&nbsp;Yoshiki Takeoka,&nbsp;Takashi Yurube,&nbsp;Takeru Tsujimoto,&nbsp;Yutaro Kanda,&nbsp;Kunihiko Miyazaki,&nbsp;Hiroki Ohnishi,&nbsp;Masao Ryu,&nbsp;Naotoshi Kumagai,&nbsp;Kohei Kuroshima,&nbsp;Yoshiaki Hiranaka,&nbsp;Ryosuke Kuroda,&nbsp;Kenichiro Kakutani","doi":"10.1002/jsp2.70046","DOIUrl":"https://doi.org/10.1002/jsp2.70046","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Transient receptor potential vanilloid 4 (TRPV4) has been identified as a Ca²⁺-permeable channel and is activated under physiological mechanical stimulation in disc nucleus pulposus (NP) cells. Meanwhile, the Ca²⁺-dependent AMP-activated protein kinase (AMPK)/mTOR pathway activates autophagy in notochordal cells. We hypothesized that TRPV4 is involved in the maintenance of intradiscal homeostasis via autophagy. Our objective was to elucidate the role of TRPV4 in extracellular matrix (ECM) metabolism and autophagy in the rat intervertebral disc through a loss-of-function study with the RNA interference (RNAi) technique.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In vitro study: Small interfering RNA (siRNA) was applied to knockdown TRPV4 by the reverse transfection method in rat disc NP cells. Expression of TRPV4, AMPK/mTOR pathway-related markers, and autophagy markers were measured by Western blotting (WB). Next, ECM metabolism was assessed under serum starvation and/or proinflammatory interleukin-1 beta (IL-1β) stimulation. In vivo study: TRPV4 and control siRNAs were injected into rat discs. To confirm in vivo transfection, WB for TRPV4 was conducted in rat disc NP-tissue protein extracts 2, 28, and 56 days after injection. Furthermore, 24-h temporary static compression-induced disruption of TRPV4 siRNA-injected discs was observed by radiography, histomorphology, and immunofluorescence.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>In vitro study: In disc cells, three different TRPV4 siRNAs consistently suppressed autophagy with TRPV4 protein knockdown (mean 33.2% [95% CI: −50.8, −15.5], 44.1% [−61.7, −26.4], 58.3% [−76.0, −40.7]). ECM metabolism was significantly suppressed by TRPV4 RNAi under proinflammatory IL-1β stimulation. In vivo study: The WB displayed sustained decreases in TRPV4 protein expression 2, 28, and 56 days after injection. Under the loaded condition, TRPV4 siRNA-injected discs presented radiographic height loss ([−31.7, −7.75]), histomorphological damage ([0.300, 4.70]), and immunofluorescent suppression of autophagy ([1.61, 20.5]) and ECM metabolism ([−25.2, −6.41]) compared to control siRNA-injected discs at 56 days.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The TRPV4 could be a therapeutic target for intervertebral disc diseases via modulating autophagy.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143431573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}