Migrating Progenitor Cells Derived From Injured Cartilage Surface Respond to Damage-Associated Molecular Patterns

Cartilage ◽  
2021 ◽  
pp. 194760352110495
Author(s):  
Lei Ding ◽  
Cheng Zhou ◽  
Hongjun Zheng ◽  
Quanming Wang ◽  
Haiyan Song ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Progenitor Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Critical Role ◽  
Cartilage Degeneration ◽  
Expression Levels ◽  
Chondrogenic Progenitor Cells ◽  
The Difference ◽  
Molecular Patterns ◽  
Damage Associated Molecular Patterns

Objective: To delineate the response of migrating chondrogenic progenitor cells (CPCs) that arose from the surface of mechanically injured articular cartilage to proinflammatory damage-associated-molecular-patterns (DAMPs). Design: Bovine CPCs and non-CPC chondrocytes isolated from either impacted or scratched articular cartilage were studied. Those 2 types of cells were treated with mitochondrial DAMPs (MTDs; 10 nM fMLF and 10 µg/mL CpG DNA), or 10 nM HMGB1, or 10 ng/mL IL-1b for 24 hours. At the end of experiments, conditioned media and cell lysates were collected for analysis of expression levels of matrix metalloproteinases (MMPs), chemokines, and cytokines that are associated with cartilage degeneration with Western blotting and quantitative polymerase chain reaction. The difference of expression levels was compared by Welch’s t-test. Results: Our data indicated that HMGB1 and MTDs remarkably upregulated pro-MMP-13 expression in CPCs. Compared with non-CPCs, CPCs expressed significantly more baseline mRNAs of MMP-13, CXCL12, and IL-6. MTDs greatly increased the expression of MMP-13 and IL-6 in CPCs by over 100-fold ( P < 0.001). MTDs also significantly increased IL-8 expression in CPCs to a similar extent ( P < 0.001). However, when IL-1b was present, CPCs expressed less MMP-3 and active MMP-13 proteins as well as less CCL2 and IL-6 than did non-CPCs. Conclusions: We concluded that CPCs were more sensitive than non-CPCs in response to DAMPs, especially MTDs. The proinflammatory nature of CPCs implied their critical role in the early phase of posttraumatic osteoarthritis development.

Progenitor Cells Derived From Injured Cartilage Surface Respond to Damage Associated Molecular Patterns

2020 ◽  
Author(s):  
Lei Ding ◽  
Cheng Zhou ◽  
Hongjun Zheng ◽  
Quanming Wang ◽  
Haiyan Song ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Critical Role ◽  
Cartilage Degeneration ◽  
Mrna Levels ◽  
Cartilage Surface ◽  
Cpg Dna ◽  
Expression Levels ◽  
Chondrogenic Progenitor Cells ◽  
Molecular Patterns ◽  
Damage Associated Molecular Patterns

Abstract Objective: To elucidate how chondrogenic progenitor cells (CPCs) originated from mechanically injured cartilage surface respond to proinflammatory endogenous damage-associated-molecular-patterns (DAMPs). Design: Passage 1 bovine CPCs and non-CPCs isolated from injured articular cartilage either by blunt impaction or by scratches were treated with mitochondrial DAMPs (MTDs) composed of fMLF and CpG DNA, or HMGB1 (a nuclear DAMP), or IL-1b for 24 hrs. At the end of the experiments, the expression levels of matrix metalloproteinases (MMPs), chemokines, and cytokines that are associated with cartilage degeneration was examined with Western blotting and quantitative PCR. Results: Both HMGB1 and MTDs remarkably up-regulated expression level of pro-MMP-13 protein in CPCs while showed weak effect on non-CPCs. Compared to non-CPCs, CPCs expressed significantly higher baseline mRNA levels of MMP-13, CXCL12, and IL-6. MTDs further increased the expression levels of MMP-13 and IL-6 in CPCs while HMGB1 did not show such effect. When treated with MTDs, CPCs also expressed significantly higher levels of IL-8 mRNA than did non-CPCs. However, compared to non-CPCs, CPCs expressed much lower levels of MMP-3 and active MMP-13 proteins as well as lower mRNA levels of CCL2 and IL-6 in response to IL-1b. Conclusions: CPCs were more sensitive than non-CPCs in response to DAMPs, especially MTDs, to up-regulate expression of MMP-13, IL-6 and -8. When IL-1b was present, CPCs were less responsive than non-CPCs in terms of up-regulating MMP-3, CCL2, and IL-6 expression. The proinflammatory nature of CPCs implied their critical role in the early phase of PTOA development.

scholarly journals Investigating the molecular control of deer antler extract on articular cartilage 

2020 ◽  
Author(s):  
Baojin Yao ◽  
Zhenwei Zhou ◽  
Mei Zhang ◽  
Xiangyang Leng ◽  
Daqing Zhao
Keyword(s):  
Articular Cartilage ◽  
Therapeutic Targets ◽  
Cartilage Degeneration ◽  
Susceptibility Genes ◽  
Water Soluble ◽  
Functional Genes ◽  
Deer Antler ◽  
Blank Group ◽  
Molecular Control ◽  
Expression Levels

Abstract Background: Deer antler is considered as a precious traditional Chinese medicinal material, and has been widely used to reinforce kidney’s yang, nourish essence and strengthen bone function. The most prominent bioactive components in deer antler are water-soluble proteins that play potential roles in bone formation and repair. The aim of this study was to explore the molecular control and therapeutic targets of deer antler extract (DAE) on articular cartilage.Methods: DAE was prepared as previously described. All rats were randomly divided into Blank group and DAE group (10 rats per group) after 7-day adaptive feeding. The rats in DAE group were orally administrated with DAE at a dose of 0.2 g/kg per day for 3 weeks and the rats in Blank group were fed with drinking water. Total RNA was isolated from the articular cartilage of knee joints. RNA sequencing (RNA-seq) experiment combined with quantitative real-time polymerase chain reaction (qRT-PCR) verification assay were carried out to explore the molecular control and therapeutic targets of DAE on articular cartilage.Results: We demonstrated that DAE significantly increased the expression levels of functional genes involved in cartilage formation, growth and repair, and decreased the expression levels of susceptibility genes involved in the pathophysiology of osteoarthritis. Conclusions: DAE might serve as a candidate supplement for maintaining cartilage homeostasis, and preventing cartilage degeneration and inflammation. These effects were possibly achieved by accelerating the expression of functional genes involved in chondrocyte commitment, survival, proliferation and differentiation, and suppressing the expression of susceptibility genes involved in the pathophysiology of osteoarthritis. Thus, our findings will contribute towards deepening the knowledge about the molecular control and therapeutic targets of DAE on the treatment of cartilage related diseases.

scholarly journals Recruitment of endogenous chondrogenic progenitor cells for articular cartilage repair

2015 ◽  
Vol 23 ◽  
pp. A40-A41 ◽  
Author(s):  
Y. Yu ◽  
D. Seol ◽  
B. Marc ◽  
H. Zheng ◽  
J. Buckwalter ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Progenitor Cells ◽  
Cartilage Repair ◽  
Articular Cartilage Repair ◽  
Chondrogenic Progenitor Cells

scholarly journals Osteoarthritic Subchondral Bone Release Exosomes That Promote Cartilage Degeneration

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 251
Author(s):  
Xiaoxin Wu ◽  
Ross Crawford ◽  
Yin Xiao ◽  
Xinzhan Mao ◽  
Indira Prasadam
Keyword(s):  
Gene Expression ◽  
Articular Cartilage ◽  
Rna Sequencing ◽  
Subchondral Bone ◽  
Critical Role ◽  
Cartilage Degeneration ◽  
Marker Genes ◽  
Specific Marker ◽  
Catabolic Gene ◽  
Loss Of Function

Altered subchondral bone and articular cartilage interactions have been implicated in the pathogenesis of osteoarthritis (OA); however, the mechanisms remain unknown. Exosomes are membrane-derived vesicles that have recently been recognized as important mediators of intercellular communication. Herein, we investigated if OA subchondral bone derived exosomes alter transcriptional and bioenergetic signatures of chondrocytes. Exosomes were isolated and purified from osteoblasts of nonsclerotic or sclerotic zones of human OA subchondral bone and their role on the articular cartilage chondrocytes was evaluated by measuring the extent of extracellular matrix production, cellular bioenergetics, and the expression of chondrocyte activity associated marker genes. Exosomal microRNAs were analyzed using RNA sequencing and validated by quantitative real-time PCR and loss-of-function. In coculture studies, chondrocytes internalized OA sclerotic subchondral bone osteoblast derived exosomes and triggered catabolic gene expression and reduced chondrocyte-specific marker expression a phenomenon that is often observed in OA cartilage. RNA sequencing and miRNA profiling have identified miR-210-5p, which is highly enriched in OA sclerotic subchondral bone osteoblast exosomes, triggered the catabolic gene expression in articular cartilage chondrocytes. Importantly, we demonstrate that miR-210-5p suppresses the oxygen consumption rate of chondrocytes, altering their bioenergetic state that is often observed in OA conditions. These effects were markedly inhibited by the addition of a miR-210-5p inhibitor. Our study indicates that exosomes released by OA sclerotic subchondral bone osteoblasts plays a critical role in progression of cartilage degeneration and might be a potential target for therapeutic intervention in OA.

scholarly journals m(6)A Modification of lncRNA NEAT1 Regulates Chronic Myelocytic Leukemia Progression via miR-766-5p/CDKN1A Axis

2021 ◽  
Vol 11 ◽  
Author(s):  
Fang-Yi Yao ◽  
Cui Zhao ◽  
Fang-Min Zhong ◽  
Ting-Yu Qin ◽  
Fang Wen ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Lines ◽  
Mononuclear Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Critical Role ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Aberrant Expression ◽  
Hematopoietic Stem ◽  
Expression Levels

BackgroundChronic myeloid leukemia (CML) is an acquired hematopoietic stem malignant disease originating from the myeloid system. Long non-coding RNAs (lncRNAs) have been widely explored in cancer tumorigenesis. However, their roles in CML remain largely unclear.MethodsThe peripheral blood mononuclear cells (PBMCs) and CML cell lines (K562, KCL22, MEG01, BV173) were collected for in vitro research. Real-time quantitative polymerase chain reaction was used to determine the mRNA expression levels. Cell viability and apoptosis were analyzed by cell counting kit 8 and flow cytometry assays. The targeting relationships were predicted using Starbase and TargetScan and ulteriorly verified by RNA pull-down and luciferase reporter assays. Western blotting assay was performed to assess the protein expressions. N6-methyladenosine (m6A) modification sites were predicted by SRAMP and confirmed by Methylated RNA immunoprecipitation (MeRIP) assay.ResultsLncRNA nuclear-enriched abundant transcript 1 (NEAT1) expression levels were decreased in the CML cell lines and PBMCs of CML patients. Moreover, METTL3-mediated m6A modification induced the aberrant expression of NEAT1 in CML. Overexpression of NEAT1 inhibited cell viability and promoted the apoptosis of CML cells. Additionally, miR-766-5p was upregulated in CML PBMCs and abrogated the effects of NEAT1 on cell viability and apoptosis of the CML cells. Further, CDKN1A was proved to be the target gene of miR-766-5p and was downregulated in the CML PBMCs. Knockdown of CDKN1A reversed the effects of NEAT1.ConclusionThe current research elucidates a novel METTL3/NEAT1/miR-766-5p/CDKN1A axis which plays a critical role in the progression of CML.

scholarly journals Role of Mitochondria-Associated Endoplasmic Reticulum Membrane in Inflammation-Mediated Metabolic Diseases

2016 ◽  
Vol 2016 ◽  
pp. 1-18 ◽  
Author(s):  
Themis Thoudam ◽  
Jae-Han Jeon ◽  
Chae-Myeong Ha ◽  
In-Kyu Lee
Keyword(s):  
Endoplasmic Reticulum ◽  
Signaling Pathways ◽  
Metabolic Diseases ◽  
Critical Role ◽  
Reactive Oxygen Species Generation ◽  
Inflammatory Factors ◽  
Endoplasmic Reticulum Membrane ◽  
Cellular Defense ◽  
Molecular Patterns ◽  
Damage Associated Molecular Patterns

Inflammation is considered to be one of the most critical factors involved in the development of complex metabolic diseases such as type 2 diabetes, cancer, and cardiovascular disease. A few decades ago, the discovery of mitochondria-associated endoplasmic reticulum (ER) membrane (MAM) was followed by the identification of its roles in regulating cellular homeostatic processes, ranging from cellular bioenergetics to apoptosis. MAM provides an excellent platform for numerous signaling pathways; among them, inflammatory signaling pathways associated with MAM play a critical role in cellular defense during pathogenic infections and metabolic disorders. However, induction of MAM causes deleterious effects by amplifying mitochondrial reactive oxygen species generation through increased calcium transfer from the ER to mitochondria, thereby causing mitochondrial damage and release of mitochondrial components into the cytosol as damage-associated molecular patterns (DAMPs). These mitochondrial DAMPs rapidly activate MAM-resident inflammasome components and other inflammatory factors, which promote inflammasome complex formation and release of proinflammatory cytokines in pathological conditions. Long-term stimulation of the inflammasome instigates chronic inflammation, leading to the pathogenesis of metabolic diseases. In this review, we summarize the current understanding of MAM and its association with inflammation-mediated metabolic diseases.

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