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Integrated DNA methylation analysis reveals a potential role for PTPRN2 in Marfan syndrome scoliosis DNA 甲基化综合分析揭示了 PTPRN2 在马凡氏综合征脊柱侧凸中的潜在作用。
IF 3.7 3区 医学 Q1 ORTHOPEDICS Pub Date : 2024-01-29 DOI: 10.1002/jsp2.1304
Zhen-zhong Zheng, Jing-hong Xu, Jia-lin Chen, Bin Jiang, Hong Ma, Lei Li, Ya-wei Li, Yu-liang Dai, Bing Wang

Background

Marfan syndrome (MFS) is a rare genetic disorder caused by mutations in the Fibrillin-1 gene (FBN1) with significant clinical features in the skeletal, cardiopulmonary, and ocular systems. To gain deeper insights into the contribution of epigenetics in the variability of phenotypes observed in MFS, we undertook the first analysis of integrating DNA methylation and gene expression profiles in whole blood from MFS and healthy controls (HCs).

Methods

The Illumina 850K (EPIC) DNA methylation array was used to detect DNA methylation changes on peripheral blood samples of seven patients with MFS and five HCs. Associations between methylation levels and clinical features of MFS were analyzed. Subsequently, we conducted an integrated analysis of the outcomes of the transcriptome data to analyze the correlation between differentially methylated positions (DMPs) and differentially expressed genes (DEGs) and explore the potential role of methylation-regulated DEGs (MeDEGs) in MFS scoliosis. The weighted gene co-expression network analysis was used to find gene modules with the highest correlation coefficient with target MeDEGs to annotate their functions in MFS.

Results

Our study identified 1253 DMPs annotated to 236 genes that were primarily associated with scoliosis, cardiomyopathy, and vital capacity. These conditions are typically associated with reduced lifespan in untreated MFS. We calculated correlations between DMPs and clinical features, such as cobb angle to evaluate scoliosis and FEV1% to assess pulmonary function. Notably, cg20223687 (PTPRN2) exhibited a positive correlation with cobb angle of scoliosis, potentially playing a role in ERKs inactivation.

Conclusions

Taken together, our systems-level approach sheds light on the contribution of epigenetics to MFS and offers a plausible explanation for the complex phenotypes that are linked to reduced lifespan in untreated MFS patients.

背景:马凡综合征(MFS)是由纤连蛋白-1基因(FBN1)突变引起的一种罕见遗传性疾病,在骨骼、心肺和眼部系统具有显著的临床特征。为了深入了解表观遗传学在 MFS 表型变异中的作用,我们首次对 MFS 和健康对照组(HCs)全血的 DNA 甲基化和基因表达谱进行了整合分析:方法:使用Illumina 850K (EPIC) DNA甲基化阵列检测7名MFS患者和5名健康对照者外周血样本的DNA甲基化变化。分析了甲基化水平与 MFS 临床特征之间的关联。随后,我们对转录组数据的结果进行了综合分析,分析了差异甲基化位点(DMPs)与差异表达基因(DEGs)之间的相关性,并探讨了甲基化调控的DEGs(MeDEGs)在MFS脊柱侧凸中的潜在作用。通过加权基因共表达网络分析,找到与目标 MeDEGs 相关系数最高的基因模块,以注释它们在 MFS 中的功能:我们的研究发现了1253个DMPs,注释了236个基因,这些基因主要与脊柱侧弯、心肌病和生命能力有关。这些情况通常与未经治疗的 MFS 的寿命缩短有关。我们计算了 DMP 与临床特征之间的相关性,如评估脊柱侧弯的 cobb 角和评估肺功能的 FEV1%。值得注意的是,cg20223687(PTPRN2)与脊柱侧弯的柯布角呈正相关,可能在ERKs失活中发挥作用:综上所述,我们的系统水平方法揭示了表观遗传学对 MFS 的贡献,并为 MFS 患者未经治疗而寿命缩短的复杂表型提供了合理的解释。
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引用次数: 0
Dynamics of N6-methyladenosine modification during aging and their potential roles in the degeneration of intervertebral disc 老化过程中 N6-甲基腺苷修饰的动态变化及其在椎间盘退化中的潜在作用。
IF 3.7 3区 医学 Q1 ORTHOPEDICS Pub Date : 2024-01-25 DOI: 10.1002/jsp2.1316
Libangxi Liu, Hong Sun, Yang Zhang, Chang Liu, Yong Zhuang, Miao Liu, Xuezheng Ai, Dan Long, Bo Huang, Changqing Li, Yue Zhou, Shiwu Dong, Chencheng Feng

Background

The N6-methyladenosine (m6A) dynamics in the progression of intervertebral disc (IVD) aging remain largely unknown. This study aimed to explore the distribution and pattern of m6A modification in nucleus pulpous (NP) tissues of rats at different ages.

Methods

Histological staining and MRI were performed to evaluate the degeneration of IVD. The expression of m6A modifiers was analyzed using qRT-PCR and western blot. Subsequently, methylated RNA immunoprecipitation next generation sequencing and RNA-seq were conducted to identify differences in m6A methylome and transcriptome of NP tissues.

Results

Compared to 2-month-old rats, we found significant changes in the global m6A level and the expression of Mettl3 and FTO in NP tissues from 20-month-old rats. During the progression of NP aging, there were 1126 persistently differentially m6A peaks within 931 genes, and 51 persistently differentially expressed genes. GO and KEGG analyses showed that these m6A peaks and m6A modified genes were mainly engaged in the biological processes and pathways of intervertebral disc degermation (IDD), such as extracellular matrix metabolism, angiogenesis, inflammatory response, mTOR and AMPK signaling pathways. Meanwhile, conjoint analyses and Venn diagram revealed a total of 405 aging related genes contained significant methylation and expression levels in 20-month-old rats in contrast to 2-month-old and 10-month-old rats. Moreover, it was found that four aging related genes with hypermethylated modification including BUB1, CA12, Adamts1, and Adamts4 depicted differentially expressed at protein level, of which BUB1 and CA12 were decreased, while Adamts1 and Adamts4 were increased during the progression of NP aging.

Conclusion

Collectively, this study elucidated the distribution and pattern of m6A modification during the aging of IVD. Furthermore, the m6A modified genes were involved in the IDD related biological processes and pathways. These findings may provide novel insights into the mechanisms and therapies of IDD from the perspective of aging.

背景:N6-甲基腺苷(m6A)在椎间盘(IVD)老化过程中的动态变化在很大程度上仍是未知的。本研究旨在探讨不同年龄大鼠髓核组织中 m6A 修饰物的分布和模式。采用 qRT-PCR 和 Western 印迹法分析 m6A修饰物的表达。随后,进行了甲基化 RNA 免疫沉淀新一代测序和 RNA-seq 分析,以确定 NP 组织中 m6A 甲基组和转录组的差异:结果:与 2 月龄大鼠相比,我们发现 20 月龄大鼠 NP 组织中 m6A 的整体水平以及 Mettl3 和 FTO 的表达均发生了显著变化。在NP衰老过程中,931个基因中有1126个持续差异m6A峰,51个持续差异表达基因。GO和KEGG分析表明,这些m6A峰和m6A修饰基因主要参与椎间盘退化(IDD)的生物学过程和通路,如细胞外基质代谢、血管生成、炎症反应、mTOR和AMPK信号通路等。同时,联合分析和维恩图显示,20月龄大鼠与2月龄和10月龄大鼠相比,共有405个衰老相关基因含有显著的甲基化和表达水平。此外,研究还发现,BUB1、CA12、Adamts1和Adamts4等4个存在高甲基化修饰的衰老相关基因在蛋白水平上存在差异表达,其中BUB1和CA12在NP衰老过程中表达量减少,而Adamts1和Adamts4在NP衰老过程中表达量增加:总之,本研究阐明了 IVD 衰老过程中 m6A 修饰的分布和模式。此外,m6A修饰基因参与了IDD相关的生物学过程和通路。这些发现可能会从衰老的角度为 IDD 的机制和治疗提供新的见解。
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引用次数: 0
Developmental morphogens direct human induced pluripotent stem cells toward an annulus fibrosus-like cell phenotype 发育形态原引导人类诱导多能干细胞形成环状纤维样细胞表型。
IF 3.7 3区 医学 Q1 ORTHOPEDICS Pub Date : 2024-01-25 DOI: 10.1002/jsp2.1313
Ana P. Peredo, Tonia K. Tsinman, Edward D. Bonnevie, Xi Jiang, Harvey E. Smith, Sarah E. Gullbrand, Nathaniel A. Dyment, Robert L. Mauck
<div> <section> <h3> Introduction</h3> <p>Therapeutic interventions for intervertebral disc herniation remain scarce due to the inability of endogenous annulus fibrosus (AF) cells to respond to injury and drive tissue regeneration. Unlike other orthopedic tissues, such as cartilage, delivery of exogenous cells to the site of annular injury remains underdeveloped, largely due to a lack of an ideal cell source and the invasive nature of cell isolation. Human induced pluripotent stem cells (iPSCs) can be differentiated to specific cell fates using biochemical factors and are, therefore, an invaluable tool for cell therapy approaches. While differentiation protocols have been developed for cartilage and fibrous connective tissues (e.g., tendon), the signals that regulate the induction and differentiation of human iPSCs toward the AF fate remain unknown.</p> </section> <section> <h3> Methods</h3> <p>iPSC-derived sclerotome cells were treated with various combinations of developmental signals including transforming growth factor beta 3 (TGF-β3), connective tissue growth factor (CTGF), platelet derived growth factor BB (PDGF-BB), insulin-like growth factor 1 (IGF-1), or the Hedgehog pathway activator, Purmorphamine, and gene expression changes in major AF-associated ECM genes were assessed. The top performing combination treatments were further validated by using three distinct iPSC lines and by assessing the production of upregulated ECM proteins of interest. To conduct a broader analysis of the transcriptomic shifts elicited by each factor combination, and to compare genetic profiles of treated cells to mature human AF cells, a 96.96 Fluidigm gene expression array was applied, and principal component analysis was employed to identify the transcriptional signatures of each cell population and treatment group in comparison to native AF cells.</p> </section> <section> <h3> Results</h3> <p>TGF-β3, in combination with PDGF-BB, CTGF, or IGF-1, induced an upregulation of key AF ECM genes in iPSC-derived sclerotome cells. In particular, treatment with a combination of TGF-β3 with PDGF-BB for 14 days significantly increased gene expression of collagen II and aggrecan and increased protein deposition of collagen I and elastin compared to other treatment groups. Assessment of genes uniquely highly expressed by AF cells or SCL cells, respectively, revealed a shift toward the genetic profile of AF cells with the addition of TGF-β3 and PDGF-BB for 14 days.</p> </section> <section> <h3> Discussion</h3> <p>These findings represent an initial approach to guide human ind
导言:由于内源性纤维环(AF)细胞无法对损伤做出反应并驱动组织再生,因此治疗椎间盘突出症的干预措施仍然很少。与软骨等其他骨科组织不同,将外源性细胞输送到椎间盘环损伤部位的技术仍不发达,这主要是由于缺乏理想的细胞来源以及细胞分离的侵入性。人类诱导多能干细胞(iPSC)可利用生化因子分化为特定的细胞命运,因此是细胞疗法的宝贵工具。虽然已开发出软骨和纤维结缔组织(如肌腱)的分化方案,但调节软骨和纤维结缔组织分化的信号还不明确、肌腱)的分化方案,但调节人类 iPSCs 向房颤命运诱导和分化的信号仍然未知。方法:用各种发育信号组合处理 iPSC 衍生的硬骨细胞,包括转化生长因子 beta 3(TGF-β3)、结缔组织生长因子(CTGF)、血小板衍生生长因子 BB(PDGF-BB)、胰岛素样生长因子 1(IGF-1)或刺猬通路激活剂 Purmorphamine,并评估主要 AF 相关 ECM 基因的基因表达变化。通过使用三种不同的 iPSC 株系和评估上调的相关 ECM 蛋白的生成情况,进一步验证了表现最佳的组合疗法。为了对每种因子组合引起的转录组变化进行更广泛的分析,并将处理过的细胞的基因图谱与成熟的人类房颤细胞进行比较,应用了96.96 Fluidigm基因表达阵列,并采用主成分分析法确定了每个细胞群和处理组与原生房颤细胞相比的转录特征:结果:TGF-β3 与 PDGF-BB、CTGF 或 IGF-1 结合使用可诱导 iPSC 衍生的硬骨体细胞中的关键 AF ECM 基因上调。特别是,与其他处理组相比,TGF-β3 与 PDGF-BB 联合处理 14 天可显著增加胶原蛋白 II 和凝集素的基因表达,并增加胶原蛋白 I 和弹性蛋白的蛋白质沉积。对AF细胞或SCL细胞分别表达的独特高表达基因的评估显示,在添加TGF-β3和PDGF-BB 14天后,AF细胞的基因特征发生了转变:这些发现代表了一种引导人类诱导多能干细胞向AF样命运转变的初步方法,可用于细胞递送策略。
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引用次数: 0
Transcriptomic profiling reveals key early response genes during GDF6-mediated differentiation of human adipose-derived stem cells to nucleus pulposus cells 转录组分析揭示了 GDF6 介导的人脂肪干细胞向髓核细胞分化过程中的关键早期反应基因
IF 3.7 3区 医学 Q1 ORTHOPEDICS Pub Date : 2024-01-19 DOI: 10.1002/jsp2.1315
Hamish T. J. Gilbert, Francis E. J. Wignall, Leo Zeef, Judith A. Hoyland, Stephen M. Richardson

Background

Stem cell-based therapies show promise as a means of repairing the degenerate intervertebral disc, with growth factors often used alongside cells to help direct differentiation toward a nucleus pulposus (NP)-like phenotype. We previously demonstrated adipose-derived stem cell (ASC) differentiation with GDF6 as optimal for generating NP-like cells through evaluating end-stage differentiation parameters. Here we conducted a time-resolved transcriptomic characterization of ASCs response to GDF6 stimulation to understand the early drivers of differentiation to NP-like cells.

Methods

Human ASCs were treated with recombinant human GDF6 for 2, 6, and 12 h. RNA sequencing and detailed bioinformatic analysis were used to assess differential gene expression, gene ontology (GO), and transcription factor involvement during early differentiation. Quantitative polymerase chain reaction (qPCR) was used to validate RNA sequencing findings and inhibitors used to interrogate Smad and Erk signaling pathways, as well as identify primary and secondary response genes.

Results

The transcriptomic response of ASCs to GDF6 stimulation was time-resolved and highly structured, with “cell differentiation” “developmental processes,” and “response to stimulus” identified as key biological process GO terms. The transcription factor ERG1 was identified as a key early response gene. Temporal cluster analysis of differentiation genes identified positive regulation NP cell differentiation, as well as inhibition of osteogenesis and adipogenesis. A role for Smad and Erk signaling in the regulation of GDF6-induced early gene expression response was observed and both primary and secondary response genes were identified.

Conclusions

This study identifies a multifactorial early gene response that contributes to lineage commitment, with the identification of a number of potentially useful early markers of differentiation of ASCs to NP cells. This detailed insight into the molecular processes in response to GDF6 stimulation of ASCs is important for the development of an efficient and efficacious cell-based therapy for intervertebral disc degeneration-associated back pain.

背景 基于干细胞的疗法有望成为修复退化椎间盘的一种手段,通常与细胞一起使用的生长因子有助于引导细胞向髓核(NP)样表型分化。我们之前通过评估末期分化参数,证明了脂肪源性干细胞(ASC)与GDF6的分化是生成NP样细胞的最佳选择。在此,我们对ASCs对GDF6刺激的反应进行了时间分辨转录组学表征,以了解向NP样细胞分化的早期驱动因素。 方法 用重组人 GDF6 处理人 ASCs 2、6 和 12 小时。使用 RNA 测序和详细的生物信息学分析来评估早期分化过程中的不同基因表达、基因本体(GO)和转录因子参与情况。定量聚合酶链反应(qPCR)用于验证 RNA 测序结果,抑制剂用于检测 Smad 和 Erk 信号通路,以及鉴定主要和次要反应基因。 结果 ASCs 对 GDF6 刺激的转录组反应具有时间分辨性和高度结构性,"细胞分化"、"发育过程 "和 "对刺激的反应 "被确定为关键生物过程 GO 术语。转录因子 ERG1 被确定为关键的早期反应基因。分化基因的时间聚类分析确定了对 NP 细胞分化的正向调控,以及对成骨和成脂的抑制。观察到 Smad 和 Erk 信号在调控 GDF6 诱导的早期基因表达反应中的作用,并确定了一级和二级反应基因。 结论 本研究发现了一种多因素的早期基因应答,这种应答有助于细胞系的承诺,同时还发现了一些潜在的有用的早期标志物,这些标志物可促进 ASCs 向 NP 细胞的分化。详细了解GDF6刺激ASCs的分子反应过程,对于开发高效、有效的细胞疗法治疗椎间盘变性相关背痛非常重要。
{"title":"Transcriptomic profiling reveals key early response genes during GDF6-mediated differentiation of human adipose-derived stem cells to nucleus pulposus cells","authors":"Hamish T. J. Gilbert,&nbsp;Francis E. J. Wignall,&nbsp;Leo Zeef,&nbsp;Judith A. Hoyland,&nbsp;Stephen M. Richardson","doi":"10.1002/jsp2.1315","DOIUrl":"https://doi.org/10.1002/jsp2.1315","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Stem cell-based therapies show promise as a means of repairing the degenerate intervertebral disc, with growth factors often used alongside cells to help direct differentiation toward a nucleus pulposus (NP)-like phenotype. We previously demonstrated adipose-derived stem cell (ASC) differentiation with GDF6 as optimal for generating NP-like cells through evaluating end-stage differentiation parameters. Here we conducted a time-resolved transcriptomic characterization of ASCs response to GDF6 stimulation to understand the early drivers of differentiation to NP-like cells.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Human ASCs were treated with recombinant human GDF6 for 2, 6, and 12 h. RNA sequencing and detailed bioinformatic analysis were used to assess differential gene expression, gene ontology (GO), and transcription factor involvement during early differentiation. Quantitative polymerase chain reaction (qPCR) was used to validate RNA sequencing findings and inhibitors used to interrogate Smad and Erk signaling pathways, as well as identify primary and secondary response genes.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The transcriptomic response of ASCs to GDF6 stimulation was time-resolved and highly structured, with “cell differentiation” “developmental processes,” and “response to stimulus” identified as key biological process GO terms. The transcription factor ERG1 was identified as a key early response gene. Temporal cluster analysis of differentiation genes identified positive regulation NP cell differentiation, as well as inhibition of osteogenesis and adipogenesis. A role for Smad and Erk signaling in the regulation of GDF6-induced early gene expression response was observed and both primary and secondary response genes were identified.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This study identifies a multifactorial early gene response that contributes to lineage commitment, with the identification of a number of potentially useful early markers of differentiation of ASCs to NP cells. This detailed insight into the molecular processes in response to GDF6 stimulation of ASCs is important for the development of an efficient and efficacious cell-based therapy for intervertebral disc degeneration-associated back pain.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"7 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.1315","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139494423","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
Repetitive strikes loading organ culture model to investigate the biological and biomechanical responses of the intervertebral disc 研究椎间盘的生物和生物力学反应的重复冲击加载器官培养模型
IF 3.7 3区 医学 Q1 ORTHOPEDICS Pub Date : 2024-01-19 DOI: 10.1002/jsp2.1314
Jiaxiang Zhou, Jianmin Wang, Jianfeng Li, Zhengya Zhu, Zhongyuan He, Junhong Li, Tao Tang, Hongkun Chen, Yukun Du, Zhen Li, Manman Gao, Zhiyu Zhou, Yongming Xi

Background

Disc degeneration is associated with repetitive violent injuries. This study aims to explore the impact of repetitive strikes loading on the biology and biomechanics of intervertebral discs (IVDs) using an organ culture model.

Methods

IVDs from the bovine tail were isolated and cultured in a bioreactor, with exposure to various loading conditions. The control group was subjected to physiological loading, while the model group was exposed to either one strike loading (compression at 38% of IVD height) or repetitive one strike loading (compression at 38% of IVD height). Disc height and dynamic compressive stiffness were measured after overnight swelling and loading. Furthermore, histological morphology, cell viability, and gene expression were analyzed on Day 32. Glycosaminoglycan (GAG) and nitric oxide (NO) release in conditioned medium were also analyzed.

Results

The repetitive one strike group exhibited early disc degeneration, characterized by decreased dynamic compression stiffness, the presence of annulus fibrosus clefts, and degradation of the extracellular matrix. Additionally, this group demonstrated significantly higher levels of cell death (p < 0.05) and glycosaminoglycan (GAG) release (p < 0.05) compared to the control group. Furthermore, upregulation of MMP1, MMP13, and ADAMTS5 was observed in both nucleus pulposus (NP) and annulus fibrosus (AF) tissues of the repetitive one strike group (p < 0.05). The one strike group exhibited annulus fibrosus clefts but showed no gene expression changes compared to the control group.

Conclusions

This study shows that repetitive violent injuries lead to the degeneration of a healthy bovine IVDs, thereby providing new insights into early-stage disc degeneration.

背景椎间盘退化与重复性暴力损伤有关。本研究旨在利用器官培养模型探讨重复性打击负荷对椎间盘(IVD)的生物学和生物力学的影响。 方法 在生物反应器中分离和培养牛尾椎间盘,并将其置于各种加载条件下。对照组承受生理负荷,而模型组则承受单击负荷(以 IVD 高度的 38% 压缩)或重复单击负荷(以 IVD 高度的 38% 压缩)。经过一夜的膨胀和加载后,对椎间盘高度和动态压缩刚度进行了测量。此外,第 32 天还对组织形态、细胞活力和基因表达进行了分析。此外,还分析了条件培养基中糖胺聚糖(GAG)和一氧化氮(NO)的释放情况。 结果 重复一击组表现出早期椎间盘退变,其特征是动态压缩硬度下降、出现纤维环裂隙和细胞外基质降解。此外,与对照组相比,该组的细胞死亡水平(p < 0.05)和糖胺聚糖(GAG)释放水平(p < 0.05)明显更高。此外,在重复一击组的髓核(NP)和环状纤维(AF)组织中都观察到了 MMP1、MMP13 和 ADAMTS5 的上调(p < 0.05)。与对照组相比,一击组出现环状纤维裂隙,但基因表达无变化。 结论 本研究表明,重复性暴力损伤会导致健康牛 IVD 退化,从而为早期椎间盘退化提供了新的见解。
{"title":"Repetitive strikes loading organ culture model to investigate the biological and biomechanical responses of the intervertebral disc","authors":"Jiaxiang Zhou,&nbsp;Jianmin Wang,&nbsp;Jianfeng Li,&nbsp;Zhengya Zhu,&nbsp;Zhongyuan He,&nbsp;Junhong Li,&nbsp;Tao Tang,&nbsp;Hongkun Chen,&nbsp;Yukun Du,&nbsp;Zhen Li,&nbsp;Manman Gao,&nbsp;Zhiyu Zhou,&nbsp;Yongming Xi","doi":"10.1002/jsp2.1314","DOIUrl":"https://doi.org/10.1002/jsp2.1314","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Disc degeneration is associated with repetitive violent injuries. This study aims to explore the impact of repetitive strikes loading on the biology and biomechanics of intervertebral discs (IVDs) using an organ culture model.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>IVDs from the bovine tail were isolated and cultured in a bioreactor, with exposure to various loading conditions. The control group was subjected to physiological loading, while the model group was exposed to either one strike loading (compression at 38% of IVD height) or repetitive one strike loading (compression at 38% of IVD height). Disc height and dynamic compressive stiffness were measured after overnight swelling and loading. Furthermore, histological morphology, cell viability, and gene expression were analyzed on Day 32. Glycosaminoglycan (GAG) and nitric oxide (NO) release in conditioned medium were also analyzed.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The repetitive one strike group exhibited early disc degeneration, characterized by decreased dynamic compression stiffness, the presence of annulus fibrosus clefts, and degradation of the extracellular matrix. Additionally, this group demonstrated significantly higher levels of cell death (<i>p</i> &lt; 0.05) and glycosaminoglycan (GAG) release (<i>p</i> &lt; 0.05) compared to the control group. Furthermore, upregulation of MMP1, MMP13, and ADAMTS5 was observed in both nucleus pulposus (NP) and annulus fibrosus (AF) tissues of the repetitive one strike group (<i>p</i> &lt; 0.05). The one strike group exhibited annulus fibrosus clefts but showed no gene expression changes compared to the control group.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This study shows that repetitive violent injuries lead to the degeneration of a healthy bovine IVDs, thereby providing new insights into early-stage disc degeneration.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"7 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.1314","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139494424","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
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 ORTHOPEDICS 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。
{"title":"Rapamycin mitigates inflammation-mediated disc matrix homeostatic imbalance by inhibiting mTORC1 and inducing autophagy through Akt activation","authors":"Takashi Yurube,&nbsp;William J. Buchser,&nbsp;Zhongying Zhang,&nbsp;Prashanta Silwal,&nbsp;Michael T. Lotze,&nbsp;James D. Kang,&nbsp;Gwendolyn A. Sowa,&nbsp;Nam V. Vo","doi":"10.1002/jsp2.1303","DOIUrl":"10.1002/jsp2.1303","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>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.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>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.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>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).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>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.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"7 1","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.1303","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139390513","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
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 ORTHOPEDICS 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。
{"title":"Finite element modeling to predict the influence of anatomic variation and implant placement on performance of biological intervertebral disc implants","authors":"Maho Koga,&nbsp;Byumsu Kim,&nbsp;Marianne Lintz,&nbsp;Sertaç Kirnaz,&nbsp;Jacob L. Goldberg,&nbsp;Ibrahim Hussain,&nbsp;Branden Medary,&nbsp;Kathleen N. Meyers,&nbsp;Suzanne A. Maher,&nbsp;Roger Härtl,&nbsp;Lawrence J. Bonassar","doi":"10.1002/jsp2.1307","DOIUrl":"https://doi.org/10.1002/jsp2.1307","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>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.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>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.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>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.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>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.</p>\u0000 </section>\u0000 </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"6 4","pages":""},"PeriodicalIF":3.7,"publicationDate":"2023-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.1307","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139047542","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
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 ORTHOPEDICS 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 ORTHOPEDICS 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 ORTHOPEDICS 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。
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引用次数: 0
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