Contribution of IL-1β, 6 and TNF-α to the form of post-traumatic osteoarthritis induced by “idealized” anterior cruciate ligament reconstruction in a porcine model

Han, Pengfei; Wei, Lei; Duan, Zhiqing; Zhang, Zhiliang; Chen, Taoyu; Lu, Jiangong; Zhao, Ruipeng; Cao, Xiaoming; Li, Peng-cui; Lv, Zhi; Wei, Xiaochun

doi:10.1016/j.intimp.2018.10.007

Cited by 46 publications

(38 citation statements)

References 44 publications

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“…It is well-known that IL-1β, as an important inflammatory mediator, plays a crucial role in the progression of OA (Daheshia and Yao, 2008;Han et al, 2018;Jenei-Lanzl et al, 2019). In line with that knowledge, we used IL-1β stimulation of articular chondrocytes as a standard in vitro OA model to induce catabolic effects resembling metabolic changes in OA pathology.…”

Section: Introductionmentioning

confidence: 96%

hBMSC-Derived Extracellular Vesicles Attenuate IL-1β-Induced Catabolic Effects on OA-Chondrocytes by Regulating Pro-inflammatory Signaling Pathways

Stöckl

Lukas

et al. 2020

Front. Bioeng. Biotechnol.

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Background: Human bone marrow-derived mesenchymal stromal cells (hBMSCs) provide a promising therapeutic approach in the cell-based therapy of osteoarthritis (OA). However, several disadvantages evolved recently, including immune responses of the host and regulatory hurdles, making it necessary to search for alternative treatment options. Extracellular vesicles (EVs) are released by multiple cell types and tissues into the extracellular microenvironment, acting as message carriers during intercellular communication. Here, we investigate putative protective effects of hBMSC-derived EVs as a cell-free approach, on IL-1β-stimulated chondrocytes obtained from OA-patients.Methods: EVs were harvested from the cell culture supernatant of hBMSCs by a sequential ultracentrifugation process. Western blot, scanning electron microscopy (SEM), and nanoparticle tracking analysis (NTA) were performed to characterize the purified particles as EVs. Intracellular incorporation of EVs, derived from PHK26-labeled hBMSCs, was tested by adding the labeled EVs to human OA chondrocytes (OA-CH), followed by fluorescence microscopy. Chondrocytes were pre-stimulated with IL-1β for 24 h, followed by EVs treatment for 24 h. Subsequently, proliferation, apoptosis, and migration (wound healing) were analyzed via BrdU assay, caspase 3/7 assay, and scratch assay, respectively. With qRT-PCR, the relative expression level of anabolic and catabolic genes was determined. Furthermore, immunofluorescence microscopy and western blot were performed to evaluate the protein expression and phosphorylation levels of Erk1/2, PI3K/Akt, p38, TAK1, and NF-κB as components of pro-inflammatory signaling pathways in OA-CH.Results: EVs from hBMSCs (hBMSC-EVs) promote proliferation and reduce apoptosis of OA-CH and IL-1β-stimulated OA-CH. Moreover, hBMSC-EVs attenuate IL-1β-induced reduction of chondrocyte migration. Furthermore, hBMSC-EVs increase gene expression of PRG4, BCL2, and ACAN (aggrecan) and decrease gene expression of MMP13, ALPL, and IL1ß in OA-CH. Notably, COL2A1, SOX9, BCL2, ACAN, and COMP gene expression levels were significantly increased in IL-1β+ EV groups compared with those IL-1β groups without EVs, whereas the gene expression levels of COLX, IL1B, MMP13, and ALPL were significantly decreased in IL-1β+ EV groups compared to IL-1β groups without EVs. In addition, the phosphorylation status of Erk1/2, PI3K/Akt, p38, TAK1, and NF-κB signaling molecules, induced by IL-1β, is prevented by hBMSC- EVs.Conclusion: EVs derived from hBMSCs alleviated IL-1β-induced catabolic effects on OA-CH via promoting proliferation and migration and reducing apoptosis, probably via downregulation of IL-1ß-activated pro-inflammatory Erk1/2, PI3K/Akt, p38, TAK1, and NF-κB signaling pathways. EVs released from BMSCs may be considered as promising cell-free intervention strategy in cartilage regenerative medicine, avoiding several adverse effects of cell-based regenerative approaches.

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Section: Introductionmentioning

confidence: 96%

hBMSC-Derived Extracellular Vesicles Attenuate IL-1β-Induced Catabolic Effects on OA-Chondrocytes by Regulating Pro-inflammatory Signaling Pathways

Stöckl

Lukas

et al. 2020

Front. Bioeng. Biotechnol.

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“…Previous studies detected matrix metalloproteases and interleukins as major players that mediate this process within the knee joint. 20,32,51 In this study, we chose to measure IL-1 levels because IL-1 is thought to be a major component of the inflammatory cascade and subsequent catabolic response within the joint space after ACL injury. IL-1 alpha (IL-1α) has been shown to decrease chondrocyte viability and proliferation in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…By combining in vivo and ex vivo imaging with the quantification of bodily fluid biomarkers in this porcine model, we have provided a more comprehensive perspective on the in vivo biologic alterations associated with ACL injuries. The majority of in vivo studies of ACL injury report on the expression of inflammatory markers only within the synovial fluid, 20,32,51 whereas in our study we have combined the synovial fluid analysis with an analysis of systemic biomarkers in the serum and the urine. As a result, our findings reflect the quantification of these secreted factors in the serum as well as their subsequent clearance in the urine, and this may provide insight into future clinical studies aimed at noninvasively (ie, without performing an arthrocentesis procedure) assessing the metabolic state of an injured joint.…”

Section: Discussionmentioning

confidence: 99%

A Translational Porcine Model for Human Cell–Based Therapies in the Treatment of Posttraumatic Osteoarthritis After Anterior Cruciate Ligament Injury

et al. 2020

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Background: There is a high incidence of posttraumatic osteoarthritis (PTOA) after anterior cruciate ligament (ACL) injury, and these injuries represent an enormous health care economic burden. In an effort to address this unmet clinical need, there has been increasing interest in cell-based therapies. Purpose: To establish a translational large animal model of PTOA and demonstrate the feasibility of intra-articular human cell–based interventions. Study Design: Descriptive laboratory study. Methods: Nine Yucatan mini-pigs underwent unilateral ACL transection and were monitored for up to 12 weeks after injury. Interleukin 1 beta (IL-1β) levels and collagen breakdown were evaluated longitudinally using enzyme-linked immunosorbent assays of synovial fluid, serum, and urine. Animals were euthanized at 4 weeks (n = 3) or 12 weeks (n = 3) after injury, and injured and uninjured limbs underwent magnetic resonance imaging (MRI) and histologic analysis. At 2 days after ACL injury, an additional 3 animals received an intra-articular injection of 107 human bone marrow–derived mesenchymal stem cells (hBM-MSCs) combined with a fibrin carrier. These cells were labeled with the luciferase reporter gene (hBM-MSCs-Luc) as well as fluorescent markers and intracellular iron nanoparticles. These animals were euthanized on day 0 (n = 1) or day 14 (n = 2) after injection. hBM-MSC-Luc viability and localization were assessed using ex vivo bioluminescence imaging, fluorescence imaging, and MRI. Results: PTOA was detected as early as 4 weeks after injury. At 12 weeks after injury, osteoarthritis could be detected grossly as well as on histologic analysis. Synovial fluid analysis showed elevation of IL-1β shortly after ACL injury, with subsequent resolution by 2 weeks after injury. Collagen type II protein fragments were elevated in the synovial fluid and serum after injury. hBM-MSCs-Luc were detected immediately after injection and at 2 weeks after injection using fluorescence imaging, MRI, and bioluminescence imaging. Conclusion: This study demonstrates the feasibility of reproducing the chondral changes, intra-articular cytokine alterations, and body fluid biomarker findings consistent with PTOA after ACL injury in a large animal model. Furthermore, we have demonstrated the ability of hBM-MSCs to survive and express transgene within the knee joint of porcine hosts without immunosuppression for at least 2 weeks. Clinical Relevance: This model holds great potential to significantly contribute to investigations focused on the development of cell-based therapies for human ACL injury–associated PTOA in the future (see Appendix Figure A1 , available online).

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“…Each mini-pig was first anesthetized using an intramuscular injection of 125 mg of Zoletil 50 (25 mg/ml + 25 mg/ml lyophilisate and solvent) (Virbac Group, Carros, France) before ACLR surgery, as previously described [5]. The lateral position was located, and the right hind limb incision was performed with the humerus as the central reference point.…”

Section: Methodsmentioning

confidence: 99%

Proteomics and Bioinformatics Analysis of Cartilage in Post-Traumatic Osteoarthritis in a Mini-Pig Model of Anterior Cruciate Ligament Repair

Che

Gao

et al. 2020

Med Sci Monit

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Background: Although osteoarthritis (OA) is a degenerative disease that is increasingly common with age, the pathogenesis of post-traumatic OA (PTOA) is poorly understood. This study aimed to undertake proteomics and bioinformatics analysis of cartilage in PTOA in a mini-pig model of anterior cruciate ligament repair (ACLR). Material/Methods: The mini-pig model of PTOA involved autologous orthotopic ACLR. Screening and identification of differentially expressed proteins in the knee joint cartilage in the OA cartilage group were compared with the control group using tandem mass tag (TMT)-labeling liquid chromatography with tandem mass spectrometry (LC-MS-MS). A protein expression level >1.2 fold-change represented protein upregulation and <0.83 fold-change represented protein down-regulation Bioinformatics analysis included Gene Ontology (GO) functional annotation and the Kyoto Encyclopedia of Genes and Genome (KEGG) pathway enrichment analysis to determine the biological functions and pathways of proteins showing altered expression profiles associated with OA. Results: There were 2,950 proteins screened from the knee cartilage tissues of the OA model group using quantitative TMT-labeling LC-MS-MS. There were 491 proteins identified with altered expression profiles, 198 proteins were upregulated and 293 proteins were down-regulated in the OA cartilage group. GO function and KEGG pathway enrichment analysis of the 491 proteins identified their functions in cellular processes, metabolic processes, and biological regulation. Conclusions: Proteomics and bioinformatics analysis of cartilage in PTOA in a mini-pig model of ACLR identified OA-related proteins.

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Contribution of IL-1β, 6 and TNF-α to the form of post-traumatic osteoarthritis induced by “idealized” anterior cruciate ligament reconstruction in a porcine model

Cited by 46 publications

References 44 publications

hBMSC-Derived Extracellular Vesicles Attenuate IL-1β-Induced Catabolic Effects on OA-Chondrocytes by Regulating Pro-inflammatory Signaling Pathways

hBMSC-Derived Extracellular Vesicles Attenuate IL-1β-Induced Catabolic Effects on OA-Chondrocytes by Regulating Pro-inflammatory Signaling Pathways

A Translational Porcine Model for Human Cell–Based Therapies in the Treatment of Posttraumatic Osteoarthritis After Anterior Cruciate Ligament Injury

Proteomics and Bioinformatics Analysis of Cartilage in Post-Traumatic Osteoarthritis in a Mini-Pig Model of Anterior Cruciate Ligament Repair

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