JOR Spine最新文献_第7页

Rapamycin mitigates inflammation-mediated disc matrix homeostatic imbalance by inhibiting mTORC1 and inducing autophagy through Akt activation 雷帕霉素通过抑制 mTORC1 和激活 Akt 诱导自噬，缓解炎症介导的椎间盘基质平衡失调

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2024-01-02 DOI: 10.1002/jsp2.1303

Takashi Yurube, William J. Buchser, Zhongying Zhang, Prashanta Silwal, Michael T. Lotze, James D. Kang, Gwendolyn A. Sowa, Nam V. Vo

Background

Low back pain is a global health problem that originated mainly from intervertebral disc degeneration (IDD). Autophagy, negatively regulated by the phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway, prevents metabolic and degenerative diseases by removing and recycling damaged cellular components. Despite growing evidence that autophagy occurs in the intervertebral disc, the regulation of disc cellular autophagy is still poorly understood.

Methods

Annulus fibrosus (rAF) cell cultures derived from healthy female rabbit discs were used to test the effect of autophagy inhibition or activation on disc cell fate and matrix homeostasis. Specifically, different chemical inhibitors including rapamycin, 3-methyladenine, MK-2206, and PP242 were used to modulate activities of different proteins in the PI3K/Akt/mTOR signaling pathway to assess IL-1β-induced cellular senescence, apoptosis, and matrix homeostasis in rAF cells grown under nutrient-poor culture condition.

Results

Rapamycin, an inhibitor of mTOR complex 1 (mTORC1), reduced the phosphorylation of mTOR and its effector p70/S6K in rAF cell cultures. Rapamycin also induced autophagic flux as measured by increased expression of key autophagy markers, including LC3 puncta number, LC3-II expression, and cytoplasmic HMGB1 intensity and decreased p62/SQSTM1 expression. As expected, IL-1β stimulation promoted rAF cellular senescence, apoptosis, and matrix homeostatic imbalance with enhanced aggrecanolysis and MMP-3 and MMP-13 expression. Rapamycin treatment effectively mitigated IL-1β-mediated inflammatory stress changes, but these alleviating effects of rapamycin were abrogated by chemical inhibition of Akt and mTOR complex 2 (mTORC2).

Conclusions

These findings suggest that rapamycin blunts adverse effects of inflammation on disc cells by inhibiting mTORC1 to induce autophagy through the PI3K/Akt/mTOR pathway that is dependent on Akt and mTORC2 activities. Hence, our findings identify autophagy, rapamycin, and PI3K/Akt/mTOR signaling as potential therapeutic targets for IDD treatment.

腰痛是一个全球性的健康问题，主要源于椎间盘退变（IDD）。自噬受磷脂酰肌醇 3- 激酶（PI3K）/Akt/哺乳动物雷帕霉素靶标（mTOR）信号通路的负调控，通过清除和回收受损细胞成分来预防代谢性和退行性疾病。尽管越来越多的证据表明自噬发生在椎间盘中，但人们对椎间盘细胞自噬的调控仍然知之甚少。我们用来自健康雌兔椎间盘的纤维环（rAF）细胞培养物来测试自噬抑制或激活对椎间盘细胞命运和基质平衡的影响。具体来说，研究人员使用不同的化学抑制剂（包括雷帕霉素、3-甲基腺嘌呤、MK-2206和PP242）来调节PI3K/Akt/mTOR信号通路中不同蛋白的活性，以评估IL-1β诱导的细胞衰老、凋亡和在营养缺乏培养条件下生长的rAF细胞的基质稳态。雷帕霉素是mTOR复合体1（mTORC1）的抑制剂，它能减少rAF细胞培养物中mTOR及其效应因子p70/S6K的磷酸化。雷帕霉素还能诱导自噬通量，其测量方法是增加关键自噬标记物的表达，包括 LC3 点的数量、LC3-II 的表达和细胞质 HMGB1 的强度，以及降低 p62/SQSTM1 的表达。正如预期的那样，IL-1β的刺激促进了rAF细胞的衰老、凋亡和基质平衡失调，增强了凝集素溶解、MMP-3和MMP-13的表达。这些研究结果表明，雷帕霉素通过抑制mTORC1，通过依赖于Akt和mTORC2活性的PI3K/Akt/mTOR途径诱导自噬，从而减轻炎症对椎间盘细胞的不利影响。因此，我们的研究结果将自噬、雷帕霉素和PI3K/Akt/mTOR信号转导确定为治疗IDD的潜在治疗靶点。

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引用次数: 0

Finite element modeling to predict the influence of anatomic variation and implant placement on performance of biological intervertebral disc implants 通过有限元建模预测解剖变异和植入物位置对生物椎间盘植入物性能的影响

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-27 DOI: 10.1002/jsp2.1307

Maho Koga, Byumsu Kim, Marianne Lintz, Sertaç Kirnaz, Jacob L. Goldberg, Ibrahim Hussain, Branden Medary, Kathleen N. Meyers, Suzanne A. Maher, Roger Härtl, Lawrence J. Bonassar

Background

Tissue-engineered intervertebral disc (TE-IVD) constructs are an attractive therapy for treating degenerative disc disease and have previously been investigated in vivo in both large and small animal models. The mechanical environment of the spine is notably challenging, in part due to its complex anatomy, and implants may require additional mechanical support to avoid failure in the early stages of implantation. As such, the design of suitable support implants requires rigorous validation.

Methods

We created a FE model to simulate the behavior of the IVD cages under compression specific to the anatomy of the porcine cervical spine, validated the FE model using an animal model, and predicted the effects of implant location and vertebral angle of the motion segment on implant behavior. Specifically, we tested anatomical positioning of the superior vertebra and placement of the implant. We analyzed corresponding stress and strain distributions.

Results

Results demonstrated that the anatomical geometry of the porcine cervical spine led to concentrated stress and strain on the posterior side of the cage. This stress concentration was associated with the location of failure of the cages reported in vivo, despite superior mechanical properties of the implant. Furthermore, placement of the cage was found to have profound effects on migration, while the angle of the superior vertebra affected stress concentration of the cage.

Conclusions

This model can be utilized both to inform surgical procedures and provide insight on future cage designs and can be adopted to models without the use of in vivo animal models.

背景组织工程椎间盘（TE-IVD）结构是治疗椎间盘退行性疾病的一种极具吸引力的疗法，以前曾在大型和小型动物模型中进行过体内研究。脊柱的机械环境具有明显的挑战性，部分原因是其复杂的解剖结构，植入物可能需要额外的机械支撑，以避免在植入初期出现故障。因此，设计合适的支撑植入物需要严格的验证。方法我们创建了一个有限元模型来模拟 IVD 骨架在猪颈椎解剖结构压缩下的行为，使用动物模型验证了有限元模型，并预测了植入物位置和运动节段椎体角度对植入物行为的影响。具体来说，我们测试了上椎体的解剖定位和植入物的位置。我们分析了相应的应力和应变分布。结果结果表明，猪颈椎的解剖几何形状导致应力和应变集中在笼的后侧。尽管植入物具有优异的机械性能，但这种应力集中与体内报告的笼失效位置有关。此外，研究还发现保持架的放置位置会对移位产生深远影响，而上椎体的角度则会影响保持架的应力集中。结论该模型既可用于指导手术过程，也可为未来的骨架设计提供启示，而且无需使用体内动物模型也可采用该模型。

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引用次数: 0

Bioinformatics-based discovery of intervertebral disc degeneration biomarkers and immune-inflammatory infiltrates 基于生物信息学的椎间盘退变生物标记物和免疫炎症浸润的发现

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-22 DOI: 10.1002/jsp2.1311

Chao Song, Daqian Zhou, Kang Cheng, Fei Liu, Weiye Cai, Yongliang Mei, Jingwen Chen, Chenyi Huang, Zongchao Liu

Background

Intervertebral disc degeneration (IVDD) is a common chronic disease in orthopedics, and its molecular mechanisms are still not well explained.

Aim

This study's objective was to bioinformatics-based discovery of IVDD biomarkers and immune-inflammatory infiltrates.

Materials and Methods

The IVDD illness gene collection was gathered from GeneCards, DisGeNet, and gene expression profiles were chosen from the extensive Gene Expression Omnibus database (GSE124272, GSE150408, and GSE153761). The STRING database was used to create a network of protein–protein interactions, while the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) databases were used for functional enrichment analysis. Using hub genes, the immune cell infiltration between IVDD patient samples and control tissues was examined. Finally, quantitative polymerase chain reaction and Western blot experiments were used to verify the expression of hub genes.

Results

A total of 27 differentially expressed hub genes were identified by bioinformatics. According to GO and KEGG analyses, hub genes were prominent in immunological responses, chemokine-mediated signaling pathways, and inflammatory responses, with the key signaling pathways engaged in cellular senescence, apoptosis, Th1 and Th2 cell differentiation, and Th17 cell differentiation. Immune cell infiltration research revealed that T cells, lymphocytes, B cells, and NK cells were decreased in IVDD patients while monocytes, neutrophils, and CD8 T cells were increased. The expression levels of the senescence hub genes SP1, VEGFA, IL-6, and the apoptosis key gene CASP3 were considerably greater in the IVDD model group than in the control group, according to in vitro validation.

Conclusion

In conclusion, the cellular senescence signaling pathway, the apoptosis signaling pathway, and associated hub genes play significant roles in the development and progression of IVDD, this finding may help direct future research on the senescence signaling route in IVDD.

IVDD疾病基因收集自GeneCards和DisGeNet，基因表达谱则选自庞大的基因表达总库数据库（GSE124272、GSE150408和GSE153761）。STRING 数据库用于创建蛋白质-蛋白质相互作用网络，而京都基因和基因组百科全书（KEGG）和基因本体（GO）数据库则用于功能富集分析。利用中枢基因，研究了IVDD患者样本与对照组织之间的免疫细胞浸润情况。最后，利用定量聚合酶链反应和 Western 印迹实验验证了中枢基因的表达情况。根据GO和KEGG分析，中心基因在免疫反应、趋化因子介导的信号通路和炎症反应中表现突出，其关键信号通路参与了细胞衰老、细胞凋亡、Th1和Th2细胞分化以及Th17细胞分化。免疫细胞浸润研究显示，IVDD 患者的 T 细胞、淋巴细胞、B 细胞和 NK 细胞减少，而单核细胞、中性粒细胞和 CD8 T 细胞增加。总之，细胞衰老信号通路、细胞凋亡信号通路及相关的中枢基因在IVDD的发生和发展中起着重要作用，这一发现可能有助于指导未来对IVDD衰老信号通路的研究。

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引用次数: 0

SSR1 and CKAP4 as potential biomarkers for intervertebral disc degeneration based on integrated bioinformatics analysis 基于综合生物信息学分析的 SSR1 和 CKAP4--椎间盘退变的潜在生物标记物

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-20 DOI: 10.1002/jsp2.1309

Danqing Guo, Min Zeng, Miao Yu, Jingjing Shang, Jinxing Lin, Lichu Liu, Kuangyang Yang, Zhenglin Cao

Background

Intervertebral disc degeneration (IDD) is a significant cause of low back pain and poses a significant public health concern. Genetic factors play a crucial role in IDD, highlighting the need for a better understanding of the underlying mechanisms.

Aim

The aim of this study was to identify potential IDD-related biomarkers using a comprehensive bioinformatics approach and validate them in vitro.

Materials and Methods

In this study, we employed several analytical approaches to identify the key genes involved in IDD. We utilized weighted gene coexpression network analysis (WGCNA), MCODE, LASSO algorithms, and ROC curves to identify the key genes. Additionally, immune infiltrating analysis and a single-cell sequencing dataset were utilized to further explore the characteristics of the key genes. Finally, we conducted in vitro experiments on human disc tissues to validate the significance of these key genes in IDD.

Results

we obtained gene expression profiles from the GEO database (GSE23130 and GSE15227) and identified 1015 DEGs associated with IDD. Using WGCNA, we identified the blue module as significantly related to IDD. Among the DEGs, we identified 47 hub genes that overlapped with the genes in the blue module, based on criteria of |logFC| ≥ 2.0 and p.adj <0.05. Further analysis using both MCODE and LASSO algorithms enabled us to identify five key genes, of which CKAP4 and SSR1 were validated by GSE70362, demonstrating significant diagnostic value for IDD. Additionally, immune infiltrating analysis revealed that monocytes were significantly correlated with the two key genes. We also analyzed a single-cell sequencing dataset, GSE199866, which showed that both CKAP4 and SSR1 were highly expressed in fibrocartilage chondrocytes. Finally, we validated our findings in vitro by performing real time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) on 30 human disc samples. Our results showed that CKAP4 and SSR1 were upregulated in degenerated disc samples. Taken together, our findings suggest that CKAP4 and SSR1 have the potential to serve as disease biomarkers for IDD.

椎间盘变性（IDD）是导致腰背痛的一个重要原因，也是一个重大的公共卫生问题。本研究的目的是利用综合生物信息学方法确定潜在的 IDD 相关生物标志物，并在体外进行验证。在本研究中，我们采用了多种分析方法来确定参与 IDD 的关键基因。我们利用加权基因共表达网络分析（WGCNA）、MCODE、LASSO算法和ROC曲线来确定关键基因。此外，我们还利用免疫浸润分析和单细胞测序数据集进一步探索了关键基因的特征。最后，我们在人体椎间盘组织上进行了体外实验，以验证这些关键基因在 IDD 中的重要性。我们从 GEO 数据库（GSE23130 和 GSE15227）中获得了基因表达谱，并确定了 1015 个与 IDD 相关的 DEGs。利用 WGCNA，我们确定了与 IDD 显著相关的蓝色模块。在 DEGs 中，根据 |logFC| ≥ 2.0 和 p.adj <0.05 的标准，我们发现了 47 个与蓝色模块中的基因重叠的枢纽基因。利用 MCODE 和 LASSO 算法进行的进一步分析使我们确定了五个关键基因，其中 CKAP4 和 SSR1 通过 GSE70362 验证，对 IDD 有显著的诊断价值。此外，免疫浸润分析表明，单核细胞与这两个关键基因显著相关。我们还分析了单细胞测序数据集 GSE199866，该数据集显示 CKAP4 和 SSR1 在纤维软骨软骨细胞中均有高表达。最后，我们对 30 个人体椎间盘样本进行了实时聚合酶链反应（RT-PCR）和免疫组化（IHC），在体外验证了我们的发现。结果显示，CKAP4 和 SSR1 在退化的椎间盘样本中上调。综上所述，我们的研究结果表明，CKAP4和SSR1有可能成为IDD的疾病生物标志物。

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引用次数: 0

Threats and opportunities of using ChatGPT in scientific writing—The risk of getting spineless 在科普写作中使用 ChatGPT 的威胁与机遇--没有骨气的风险

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-13 DOI: 10.1002/jsp2.1296

Luca Ambrosio, Jordy Schol, Vincenzo Amedeo La Pietra, Fabrizio Russo, Gianluca Vadalà, Daisuke Sakai

ChatGPT and AI chatbots are revolutionizing several science fields, including medical writing. However, the inadequate use of such advantageous tools can raise numerous methodological and ethical issues.

ChatGPT 和人工智能聊天机器人正在彻底改变多个科学领域，包括医学写作。然而，如果不充分使用这些有利工具，就会引发许多方法论和伦理问题。

引用次数: 0

Links among MRI features in paraspinal muscles, inflammatory processes, and related back pain in patients with lumbar disc herniation 腰椎间盘突出症患者脊柱旁肌肉的 MRI 特征、炎症过程和相关背痛之间的联系

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-13 DOI: 10.1002/jsp2.1310

Xiaolong Chen, Peng Cui, Yongjin Li, Yu Wang, Shibao Lu

Background

Recent studies have provided evidence that structural changes in paraspinal muscles are associated with intervertebral disc degeneration (IDD), ubiquitous with low back pain (LBP), and potentially thought to be regulated by inflammatory processes. However, the links remain unclear.

Objective

The aims of this study were to investigate structural changes in paraspinal muscles that differed in healthy and lumbar disc herniation (LDH) patients, and LDH patients with and without LBP, and to determine the link with the expression of inflammatory marker(s).

Methods

Cross-sectional areas (CSAs) and fatty degeneration of muscles were measured in this prospective cohort study. Multifidus muscle (MM) tissue was procured from included individuals undergoing surgery. Gene expression was quantified using qPCR assays. Independent t-test, Chi-square, and Spearman correlation were used for evaluating the links among structural changes, expression of inflammatory markers, and clinical outcomes.

Results

Functional CSA and fatty degeneration of MM were larger in healthy group than LDH group. A significant increase in fat infiltration in MM in LBP group than in non-LBP group. TNF-alpha (TNF-α) was 28-fold greater in high-fat infiltration group than low-fat infiltration group within MM. Expression of TNF-α and IL-1β in MM was moderately correlated with functional CSA and fatty degeneration of MM, which was moderately correlated with clinical outcomes.

Conclusions

Results support the hypothesis that IDD is associated with dysregulation of inflammatory state of local MM, which provides initial evidence that inflammatory dysregulation in paraspinal muscles has the potential for a broad impact on tissue health and LBP symptoms.

最近的研究证明，脊柱旁肌肉的结构变化与椎间盘退变（IDD）有关，腰背痛（LBP）无处不在，并可能受炎症过程的调节。本研究的目的是调查健康人与腰椎间盘突出症（LDH）患者、有腰背痛与无腰背痛的腰椎间盘突出症患者脊柱旁肌肉的结构变化差异，并确定其与炎症标志物表达之间的联系。这项前瞻性队列研究对肌肉横截面积（CSA）和脂肪变性进行了测量。基因表达采用 qPCR 方法进行量化。采用独立t检验、Chi-square和Spearman相关性来评估结构变化、炎症标志物表达和临床结果之间的联系。与非 LBP 组相比，LBP 组 MM 的脂肪浸润明显增加。在 MM 中，高脂肪浸润组的 TNF-α（TNF-α）是低脂肪浸润组的 28 倍。MM中TNF-α和IL-1β的表达与MM的功能性CSA和脂肪变性呈中度相关，而MM的功能性CSA和脂肪变性与临床结果呈中度相关。研究结果支持了IDD与局部MM炎症状态失调有关的假设，这为脊柱旁肌肉的炎症失调可能对组织健康和LBP症状产生广泛影响提供了初步证据。

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引用次数: 0

Sex differences in the biomechanical and biochemical responses of caudal rat intervertebral discs to injury 尾部大鼠椎间盘对损伤的生物力学和生物化学反应的性别差异

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-09 DOI: 10.1002/jsp2.1299

Hagar M. Kenawy, María I. Nuñez, Xóchitl Morales, Lauren E. Lisiewski, Kevin G. Burt, Min Kyu M. Kim, Leonardo Campos, Nadia Kiridly, Clark T. Hung, Nadeen O. Chahine

Background

Intervertebral disc degeneration (IDD) is a major cause of low back pain (LBP) worldwide. Sexual dimorphism, or sex-based differences, appear to exist in the severity of LBP. However, it is unknown if there are sex-based differences in the inflammatory, biomechanical, biochemical, and histological responses of intervertebral discs (IVDs).

Methods

Caudal (Coccygeal/Co) bone-disc-bone motion segments were isolated from multiple spinal levels (Co8 to Co14) of male and female Sprague–Dawley rats. Changes in motion segment biomechanics and extracellular matrix (ECM) biochemistry (glycosaminoglycan [GAG], collagen [COL], water, and DNA content) were evaluated at baseline and in response to chemical insult (lipopolysaccharide [LPS]) or puncture injury ex vivo. We also investigated the contributions of Toll-like receptor (TLR4) signaling on responses to LPS or puncture injury ex vivo, using a small molecule TLR4 inhibitor, TAK-242.

Results

Findings indicate that IVD motion segments from female donors had greater nitric oxide (NO) release in LPS groups compared to male donors. HMGB1 release was increased in punctured discs, but not LPS injured discs, with no sex effect. Although both male and female discs exhibited reductions in dynamic moduli in response to LPS and puncture injuries, dynamic moduli from female donors were higher than male donors across all groups. In uninjured (baseline) samples, a significant sex effect was observed in nucleus pulposus (NP) DNA and water content. Female annulus fibrosus (AF) also had higher DNA, GAG, and COL content (normalized by dry weight), but lower water content than male AF. Additional injury- and sex-dependent effects were observed in AF GAG/DNA and COL/DNA content. Finally, TAK-242 improved the dynamic modulus of female but not male punctured discs.

Conclusions

Our findings demonstrate that there are differences in rat IVD motion segments based on sex, and that the response to injury in inflammatory, biomechanical, biochemical, and histological outcomes also exhibit sex differences. TLR4 inhibition protected against loss of mechanical integrity of puncture-injured IVD motion segments, with differences responses based on donor sex.

椎间盘退变(IDD)是世界范围内腰痛(LBP)的主要原因。性二态性，或性别差异，似乎存在于腰痛的严重程度。然而，椎间盘(ivd)的炎症、生物力学、生化和组织学反应是否存在性别差异尚不清楚。从雄性和雌性Sprague-Dawley大鼠的多个脊柱节段(Co8至Co14)中分离出尾侧(尾骨/Co)骨-盘-骨运动节段。运动段生物力学和细胞外基质(ECM)生物化学(糖胺聚糖[GAG]、胶原[COL]、水和DNA含量)的变化在基线和对化学损伤(脂多糖[LPS])或体外穿刺损伤的反应中进行了评估。我们还使用一种小分子TLR4抑制剂TAK - 242研究了Toll样受体(TLR4)信号在LPS或穿刺损伤反应中的作用。研究结果表明，与男性供者相比，LPS组女性供者的IVD运动节段有更多的一氧化氮(NO)释放。HMGB1释放在穿刺椎间盘中增加，而LPS损伤椎间盘中没有增加，无性别影响。尽管男性和女性的椎间盘在LPS和穿刺损伤后都表现出动态模量的减少，但在所有组中，女性供体的动态模量都高于男性供体。在未损伤(基线)样品中，在髓核(NP) DNA和含水量中观察到显著的性别效应。雌性纤维环(AF)也具有更高的DNA、GAG和COL含量(按干重标准化)，但含水量低于雄性纤维环。在AF中，GAG/DNA和COL/DNA含量存在额外的损伤和性别依赖效应。最后，TAK‐242改善了女性穿刺椎间盘的动态模量，但没有改善男性穿刺椎间盘的动态模量。我们的研究结果表明，基于性别的大鼠IVD运动节段存在差异，并且对炎症，生物力学，生化和组织学结果的损伤反应也表现出性别差异。TLR4抑制保护了穿刺损伤的IVD运动节段的机械完整性的丧失，基于供体性别的反应不同。

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引用次数: 0

Differential efficacy of two small molecule PHLPP inhibitors to promote nucleus Pulposus cell health 两种小分子 PHLPP 抑制剂促进细胞核健康的不同功效

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-04 DOI: 10.1002/jsp2.1306

Changli Zhang, Madeleine D. Gordon, Katherine M. Joseph, Martha E. Diaz-Hernandez, Hicham Drissi, Svenja Illien-Jünger

Background

Intervertebral disc (IVD) degeneration is associated with chronic back pain. We previously demonstrated that the phosphatase pleckstrin homology domain and leucine-rich repeat protein phosphatase (PHLPP) 1 was positively correlated with IVD degeneration and its deficiency decelerated IVD degeneration in both mouse IVDs and human nucleus pulposus (NP) cells. Small molecule PHLPP inhibitors may offer a translatable method to alleviate IVD degeneration. In this study, we tested the effectiveness of the two PHLPP inhibitors NSC117079 and NSC45586 in promoting a healthy NP phenotype.

Methods

Tail IVDs of 5-month-old wildtype mice were collected and treated with NSC117079 or NSC45586 under low serum conditions ex vivo. Hematoxylin & eosin staining was performed to examine IVD structure and NP cell morphology. The expression of KRT19 was analyzed through immunohistochemistry. Cell apoptosis was assessed by TUNEL assay. Human NP cells were obtained from patients with IVD degeneration. The gene expression of KRT19, ACAN, SOX9, and MMP13 was analyzed via real time qPCR, and AKT phosphorylation and the protein expression of FOXO1 was analyzed via immunoblot.

Results

In a mouse IVD organ culture model, NSC45586, but not NSC117079, preserved vacuolated notochordal cell morphology and KRT19 expression while suppressing cell apoptosis, counteracting the degenerative changes induced by serum deprivation, especially in males. Likewise, in degenerated human NP cells, NSC45586 increased cell viability and the expression of KRT19, ACAN, and SOX9 and reducing the expression of MMP13, while NSC117079 treatment only increased KRT19 expression. Mechanistically, NSC45586 treatment increased FOXO1 protein expression in NP cells, and inhibiting FOXO1 offset NSC45586-induced regenerative potential, especially in males.

Conclusions

Our study indicates that NSC45586 was effective in promoting NP cell health, especially in males, suggesting that PHLPP plays a key role in NP cell homeostasis and that NSC45586 might be a potential drug candidate in treating IVD degeneration.

椎间盘(IVD)退变与慢性背痛有关。我们之前已经证明，磷酸酶pleckstrin同源结构域和富含亮氨酸的重复蛋白磷酸酶(PHLPP) 1与IVD变性呈正相关，并且在小鼠IVD和人髓核(NP)细胞中，它的缺乏减缓了IVD变性。小分子PHLPP抑制剂可能提供一种可翻译的方法来减轻IVD变性。在这项研究中，我们测试了两种PHLPP抑制剂NSC117079和NSC45586在促进健康NP表型方面的有效性。收集5月龄野生型小鼠的尾ivd，在低血清条件下用NSC117079或NSC45586体外处理。苏木精和伊红染色检测IVD结构和NP细胞形态。免疫组化分析KRT19的表达。TUNEL法检测细胞凋亡。从IVD变性患者中获得人NP细胞。real - time qPCR检测KRT19、ACAN、SOX9和MMP13基因表达，免疫印迹检测AKT磷酸化和FOXO1蛋白表达。在小鼠IVD器官培养模型中，NSC45586(而非NSC117079)在抑制细胞凋亡的同时，保留了空泡化脊索细胞形态和KRT19表达，抵消了血清剥夺引起的退行性改变，尤其是在雄性小鼠中。同样，在退化的人NP细胞中，NSC45586增加了细胞活力和KRT19、ACAN和SOX9的表达，降低了MMP13的表达，而NSC117079只增加了KRT19的表达。在机制上，NSC45586处理增加了NP细胞中FOXO1蛋白的表达，抑制FOXO1抵消了NSC45586诱导的再生潜能，尤其是在雄性中。我们的研究表明，NSC45586能有效促进NP细胞健康，特别是在男性中，这表明PHLPP在NP细胞稳态中起关键作用，NSC45586可能是治疗IVD变性的潜在候选药物。

{"title":"Differential efficacy of two small molecule PHLPP inhibitors to promote nucleus Pulposus cell health","authors":"Changli Zhang, Madeleine D. Gordon, Katherine M. Joseph, Martha E. Diaz-Hernandez, Hicham Drissi, Svenja Illien-Jünger","doi":"10.1002/jsp2.1306","DOIUrl":"10.1002/jsp2.1306","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Intervertebral disc (IVD) degeneration is associated with chronic back pain. We previously demonstrated that the phosphatase pleckstrin homology domain and leucine-rich repeat protein phosphatase (PHLPP) 1 was positively correlated with IVD degeneration and its deficiency decelerated IVD degeneration in both mouse IVDs and human nucleus pulposus (NP) cells. Small molecule PHLPP inhibitors may offer a translatable method to alleviate IVD degeneration. In this study, we tested the effectiveness of the two PHLPP inhibitors NSC117079 and NSC45586 in promoting a healthy NP phenotype.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Tail IVDs of 5-month-old wildtype mice were collected and treated with NSC117079 or NSC45586 under low serum conditions ex vivo. Hematoxylin & eosin staining was performed to examine IVD structure and NP cell morphology. The expression of KRT19 was analyzed through immunohistochemistry. Cell apoptosis was assessed by TUNEL assay. Human NP cells were obtained from patients with IVD degeneration. The gene expression of KRT19, ACAN, SOX9, and MMP13 was analyzed via real time qPCR, and AKT phosphorylation and the protein expression of FOXO1 was analyzed via immunoblot.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>In a mouse IVD organ culture model, NSC45586, but not NSC117079, preserved vacuolated notochordal cell morphology and KRT19 expression while suppressing cell apoptosis, counteracting the degenerative changes induced by serum deprivation, especially in males. Likewise, in degenerated human NP cells, NSC45586 increased cell viability and the expression of KRT19, ACAN, and SOX9 and reducing the expression of MMP13, while NSC117079 treatment only increased KRT19 expression. Mechanistically, NSC45586 treatment increased FOXO1 protein expression in NP cells, and inhibiting FOXO1 offset NSC45586-induced regenerative potential, especially in males.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Our study indicates that NSC45586 was effective in promoting NP cell health, especially in males, suggesting that PHLPP plays a key role in NP cell homeostasis and that NSC45586 might be a potential drug candidate in treating IVD degeneration.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2023-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.1306","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138602428","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}

引用次数: 0

Comparative histopathological analysis of age-associated intervertebral disc degeneration in CD-1 and C57BL/6 mice: Anatomical and sex-based differences 对 CD-1 和 C57BL/6 小鼠年龄相关性椎间盘退化的组织病理学比较分析：解剖学和性别差异

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-03 DOI: 10.1002/jsp2.1298

Jeffrey L. Hutchinson, Matthew A. Veras, Meghan E. Serjeant, Matthew R. McCann, Ashley L. Kelly, Diana Quinonez, Frank Beier, Cheryle A. Séguin

Background

Intervertebral disc (IVD) degeneration is a major contributor to back pain and disability. The cause of IVD degeneration is multifactorial, with no disease-modifying treatments. Mouse models are commonly used to study IVD degeneration; however, the effects of anatomical location, strain, and sex on the progression of age-associated degeneration are poorly understood.

Methods

A longitudinal study was conducted to characterize age-, anatomical-, and sex-specific differences in IVD degeneration in two commonly used strains of mice, C57BL/6 and CD-1. Histopathological evaluation of the cervical, thoracic, lumbar, and caudal regions of mice at 6, 12, 20, and 24 months of age was conducted by two blinded observers at each IVD for the nucleus pulposus (NP), annulus fibrosus (AF), and the NP/AF boundary compartments, enabling analysis of scores by tissue compartment, summed scores for each IVD, or averaged scores for each anatomical region.

Results

C57BL/6 mice displayed mild IVD degeneration until 24 months of age; at this point, the lumbar spine demonstrated the most degeneration compared to other regions. Degeneration was detected earlier in the CD-1 mice (20 months of age) in both the thoracic and lumbar spine. In CD-1 mice, moderate to severe degeneration was noted in the cervical spine at all time points assessed. In both strains, age-associated IVD degeneration in the thoracic and lumbar spine was associated with increased histopathological scores in all IVD compartments. In both strains, minimal degeneration was detected in caudal IVDs out to 24 months of age. Both C57BL/6 and CD-1 mice displayed sex-specific differences in the presentation and progression of age-associated IVD degeneration.

Conclusions

These results showed that the progression and severity of age-associated degeneration in mouse models is associated with marked differences based on anatomical region, sex, and strain. This information provides a fundamental baseline characterization for users of mouse models to enable effective and appropriate experimental design, interpretation, and comparison between studies.

椎间盘(IVD)退变是背部疼痛和残疾的主要原因。IVD变性的原因是多因素的，没有疾病修饰治疗。常用小鼠模型研究IVD变性;然而，解剖位置、应变和性别对年龄相关退变进展的影响尚不清楚。一项纵向研究对C57BL/6和CD - 1两种常用小鼠品系的IVD变性进行了年龄、解剖和性别特异性差异的表征。在6、12、20和24月龄时，由两名盲法观察者对小鼠的颈椎、胸椎、腰椎和尾侧区域进行组织病理学评估，分别在每个IVD处对髓核(NP)、纤维环(AF)和NP/AF边界区室进行评估，从而可以按组织区室、每个IVD的总得分或每个解剖区域的平均得分进行分析。C57BL/6小鼠在24月龄前表现为轻度IVD变性;此时，与其他部位相比，腰椎表现出最严重的退变。在CD‐1小鼠(20个月大)中，在胸椎和腰椎的退变更早被检测到。在CD‐1小鼠中，在评估的所有时间点都注意到颈椎中度至重度退变。在这两个品系中，年龄相关的胸椎和腰椎IVD退变与所有IVD区室的组织病理学评分增加相关。在这两个菌株中，在24个月大的尾侧IVDs中检测到最小的变性。C57BL/6和CD - 1小鼠在年龄相关IVD变性的表现和进展方面均表现出性别特异性差异。这些结果表明，小鼠模型中年龄相关变性的进展和严重程度与基于解剖区域、性别和品系的显著差异相关。这些信息为小鼠模型的使用者提供了一个基本的基线特征，以实现有效和适当的实验设计、解释和研究之间的比较。

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引用次数: 0

Thresholding approaches for estimating paraspinal muscle fat infiltration using T1- and T2-weighted MRI: Comparative analysis using water–fat MRI 利用 T1 和 T2 加权磁共振成像估算脊柱旁肌肉脂肪浸润的阈值法：使用水脂肪磁共振成像的比较分析

IF 3.7 3区医学 Q1 Medicine

JOR Spine

Pub Date : 2023-12-01 DOI: 10.1002/jsp2.1301

Jessica Ornowski, Lucas Dziesinski, Madeline Hess, Roland Krug, Maryse Fortin, Abel Torres-Espin, Sharmila Majumdar, Valentina Pedoia, Noah B. Bonnheim, Jeannie F. Bailey

Background

Paraspinal muscle fat infiltration is associated with spinal degeneration and low back pain, however, quantifying muscle fat using clinical magnetic resonance imaging (MRI) techniques continues to be a challenge. Advanced MRI techniques, including chemical-shift encoding (CSE) based water–fat MRI, enable accurate measurement of muscle fat, but such techniques are not widely available in routine clinical practice.

Methods

To facilitate assessment of paraspinal muscle fat using clinical imaging, we compared four thresholding approaches for estimating muscle fat fraction (FF) using T1- and T2-weighted images, with measurements from water–fat MRI as the ground truth: Gaussian thresholding, Otsu's method, K-mean clustering, and quadratic discriminant analysis. Pearson's correlation coefficients (r), mean absolute errors, and mean bias errors were calculated for FF estimates from T1- and T2-weighted MRI with water–fat MRI for the lumbar multifidus (MF), erector spinae (ES), quadratus lumborum (QL), and psoas (PS), and for all muscles combined.

Results

We found that for all muscles combined, FF measurements from T1- and T2-weighted images were strongly positively correlated with measurements from the water–fat images for all thresholding techniques (r = 0.70–0.86, p < 0.0001) and that variations in inter-muscle correlation strength were much greater than variations in inter-method correlation strength.

Conclusion

We conclude that muscle FF can be quantified using thresholded T1- and T2-weighted MRI images with relatively low bias and absolute error in relation to water–fat MRI, particularly in the MF and ES, and the choice of thresholding technique should depend on the muscle and clinical MRI sequence of interest.

棘旁肌肉脂肪浸润与脊柱退变和腰痛有关，然而，使用临床磁共振成像(MRI)技术量化肌肉脂肪仍然是一个挑战。先进的MRI技术，包括基于化学移位编码(CSE)的水脂肪MRI，可以精确测量肌肉脂肪，但这些技术在常规临床实践中并不广泛使用。为了便于临床影像学评估棘旁肌脂肪，我们比较了使用T1和T2加权图像估计肌肉脂肪分数(FF)的四种阈值方法，并将水脂肪MRI测量结果作为基本事实:高斯阈值法、Otsu方法、K均值聚类和二次判别分析。用水脂肪MRI对腰多裂肌(MF)、竖脊肌(ES)、腰方肌(QL)和腰肌(PS)以及所有肌肉进行T1 -和T2 -加权MRI的FF估计，计算Pearson相关系数(r)、平均绝对误差和平均偏倚误差。我们发现，对于所有肌肉，T1和T2加权图像的FF测量值与所有阈值技术的水-脂肪图像测量值呈强正相关(r = 0.70-0.86, p < 0.0001)，肌肉间相关强度的变化远远大于方法间相关强度的变化。我们得出结论，肌肉FF可以使用阈值T1和T2加权MRI图像进行量化，相对于水脂肪MRI，特别是MF和ES，其偏差和绝对误差相对较低，阈值技术的选择应取决于感兴趣的肌肉和临床MRI序列。

{"title":"Thresholding approaches for estimating paraspinal muscle fat infiltration using T1- and T2-weighted MRI: Comparative analysis using water–fat MRI","authors":"Jessica Ornowski, Lucas Dziesinski, Madeline Hess, Roland Krug, Maryse Fortin, Abel Torres-Espin, Sharmila Majumdar, Valentina Pedoia, Noah B. Bonnheim, Jeannie F. Bailey","doi":"10.1002/jsp2.1301","DOIUrl":"10.1002/jsp2.1301","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Paraspinal muscle fat infiltration is associated with spinal degeneration and low back pain, however, quantifying muscle fat using clinical magnetic resonance imaging (MRI) techniques continues to be a challenge. Advanced MRI techniques, including chemical-shift encoding (CSE) based water–fat MRI, enable accurate measurement of muscle fat, but such techniques are not widely available in routine clinical practice.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>To facilitate assessment of paraspinal muscle fat using clinical imaging, we compared four thresholding approaches for estimating muscle fat fraction (FF) using T1- and T2-weighted images, with measurements from water–fat MRI as the ground truth: Gaussian thresholding, Otsu's method, K-mean clustering, and quadratic discriminant analysis. Pearson's correlation coefficients (<i>r</i>), mean absolute errors, and mean bias errors were calculated for FF estimates from T1- and T2-weighted MRI with water–fat MRI for the lumbar multifidus (MF), erector spinae (ES), quadratus lumborum (QL), and psoas (PS), and for all muscles combined.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>We found that for all muscles combined, FF measurements from T1- and T2-weighted images were strongly positively correlated with measurements from the water–fat images for all thresholding techniques (<i>r =</i> 0.70–0.86, <i>p <</i> 0.0001) and that variations in inter-muscle correlation strength were much greater than variations in inter-method correlation strength.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>We conclude that muscle FF can be quantified using thresholded T1- and T2-weighted MRI images with relatively low bias and absolute error in relation to water–fat MRI, particularly in the MF and ES, and the choice of thresholding technique should depend on the muscle and clinical MRI sequence of interest.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.1301","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138620759","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}

引用次数: 0