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Utility of a Mouse Model of Osteoarthritis to Demonstrate Cartilage Protection by IFN γ -Primed Equine Mesenchymal Stem Cells

View Article: PubMed Central - PubMed

ABSTRACT

Objective: Mesenchymal stem cells isolated from adipose tissue (ASC) have been shown to influence the course of osteoarthritis (OA) in different animal models and are promising in veterinary medicine for horses involved in competitive sport. The aim of this study was to characterize equine ASCs (eASCs) and investigate the role of interferon-gamma (IFNγ)-priming on their therapeutic effect in a murine model of OA, which could be relevant to equine OA.

Methods: ASC were isolated from subcutaneous fat. Expression of specific markers was tested by cytometry and RT-qPCR. Differentiation potential was evaluated by histology and RT-qPCR. For functional assays, naïve or IFNγ-primed eASCs were cocultured with peripheral blood mononuclear cells or articular cartilage explants. Finally, the therapeutic effect of eASCs was tested in the model of collagenase-induced OA (CIOA) in mice.

Results: The immunosuppressive function of eASCs on equine T cell proliferation and their chondroprotective effect on equine cartilage explants were demonstrated in vitro. Both cartilage degradation and T cell activation were reduced by naïve and IFNγ-primed eASCs, but IFNγ-priming enhanced these functions. In CIOA, intra-articular injection of eASCs prevented articular cartilage from degradation and IFNγ-primed eASCs were more potent than naïve cells. This effect was related to the modulation of eASC secretome by IFNγ-priming.

Conclusion: IFNγ-priming of eASCs potentiated their antiproliferative and chondroprotective functions. We demonstrated that the immunocompetent mouse model of CIOA was relevant to test the therapeutic efficacy of xenogeneic eASCs for OA and confirmed that IFNγ-primed eASCs may have a therapeutic value for musculoskeletal diseases in veterinary medicine.

No MeSH data available.


Effect of IFNγ-priming on the inflammatory gene profile of eASCs. Gene array analysis of inflammatory cytokines and chemokines mRNA compared naïve and IFNγ-primed eASCs. (A) Hierarchical clustering comparing naïve or IFNγ-primed eASCs, (B) induced gene expression levels in IFNγ-primed eASCs expressed as relative expression (2−ΔCT), (C) significantly modulated gene expression levels in IFNγ-primed eASCs. Results are represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Mann–Whitney test. *p < 0.05.
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Figure 5: Effect of IFNγ-priming on the inflammatory gene profile of eASCs. Gene array analysis of inflammatory cytokines and chemokines mRNA compared naïve and IFNγ-primed eASCs. (A) Hierarchical clustering comparing naïve or IFNγ-primed eASCs, (B) induced gene expression levels in IFNγ-primed eASCs expressed as relative expression (2−ΔCT), (C) significantly modulated gene expression levels in IFNγ-primed eASCs. Results are represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Mann–Whitney test. *p < 0.05.

Mentions: In order to understand the effect of IFNγ priming on the immunosuppressive properties of eASCs, a gene expression analysis was performed on 89 genes related to cytokine or chemokine families. Hierarchical clustering analysis revealed that the gene expression pattern of IFNγ-primed eASCs greatly differed from that of naïve eASCs (Figure 5A). IFNγ priming modified the transcriptomic program of eASCs by significantly dysregulating 13 genes. Gene expression of CXCL9, CXCL11-like, IL32-like, and CXCL10 was significantly induced in IFNγ-primed eASCs compared with naïve eASCs (Figure 5B). In parallel, gene expression of CSF1, IL6, IL7-like, IL-15, CCL2, CCL13, and TNFS13B was significantly upregulated in IFNγ-primed eASCs, while expression of TNFSF11-like and TGFβ2-like was downregulated as compared with naïve eASCs (Figure 5C). Indeed, the cytokine/chemokine profile of eASCs was dramatically altered by IFNγ priming.


Utility of a Mouse Model of Osteoarthritis to Demonstrate Cartilage Protection by IFN γ -Primed Equine Mesenchymal Stem Cells
Effect of IFNγ-priming on the inflammatory gene profile of eASCs. Gene array analysis of inflammatory cytokines and chemokines mRNA compared naïve and IFNγ-primed eASCs. (A) Hierarchical clustering comparing naïve or IFNγ-primed eASCs, (B) induced gene expression levels in IFNγ-primed eASCs expressed as relative expression (2−ΔCT), (C) significantly modulated gene expression levels in IFNγ-primed eASCs. Results are represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Mann–Whitney test. *p < 0.05.
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Related In: Results  -  Collection

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Show All Figures
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Figure 5: Effect of IFNγ-priming on the inflammatory gene profile of eASCs. Gene array analysis of inflammatory cytokines and chemokines mRNA compared naïve and IFNγ-primed eASCs. (A) Hierarchical clustering comparing naïve or IFNγ-primed eASCs, (B) induced gene expression levels in IFNγ-primed eASCs expressed as relative expression (2−ΔCT), (C) significantly modulated gene expression levels in IFNγ-primed eASCs. Results are represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Mann–Whitney test. *p < 0.05.
Mentions: In order to understand the effect of IFNγ priming on the immunosuppressive properties of eASCs, a gene expression analysis was performed on 89 genes related to cytokine or chemokine families. Hierarchical clustering analysis revealed that the gene expression pattern of IFNγ-primed eASCs greatly differed from that of naïve eASCs (Figure 5A). IFNγ priming modified the transcriptomic program of eASCs by significantly dysregulating 13 genes. Gene expression of CXCL9, CXCL11-like, IL32-like, and CXCL10 was significantly induced in IFNγ-primed eASCs compared with naïve eASCs (Figure 5B). In parallel, gene expression of CSF1, IL6, IL7-like, IL-15, CCL2, CCL13, and TNFS13B was significantly upregulated in IFNγ-primed eASCs, while expression of TNFSF11-like and TGFβ2-like was downregulated as compared with naïve eASCs (Figure 5C). Indeed, the cytokine/chemokine profile of eASCs was dramatically altered by IFNγ priming.

View Article: PubMed Central - PubMed

ABSTRACT

Objective: Mesenchymal stem cells isolated from adipose tissue (ASC) have been shown to influence the course of osteoarthritis (OA) in different animal models and are promising in veterinary medicine for horses involved in competitive sport. The aim of this study was to characterize equine ASCs (eASCs) and investigate the role of interferon-gamma (IFN&gamma;)-priming on their therapeutic effect in a murine model of OA, which could be relevant to equine OA.

Methods: ASC were isolated from subcutaneous fat. Expression of specific markers was tested by cytometry and RT-qPCR. Differentiation potential was evaluated by histology and RT-qPCR. For functional assays, na&iuml;ve or IFN&gamma;-primed eASCs were cocultured with peripheral blood mononuclear cells or articular cartilage explants. Finally, the therapeutic effect of eASCs was tested in the model of collagenase-induced OA (CIOA) in mice.

Results: The immunosuppressive function of eASCs on equine T cell proliferation and their chondroprotective effect on equine cartilage explants were demonstrated in vitro. Both cartilage degradation and T cell activation were reduced by na&iuml;ve and IFN&gamma;-primed eASCs, but IFN&gamma;-priming enhanced these functions. In CIOA, intra-articular injection of eASCs prevented articular cartilage from degradation and IFN&gamma;-primed eASCs were more potent than na&iuml;ve cells. This effect was related to the modulation of eASC secretome by IFN&gamma;-priming.

Conclusion: IFN&gamma;-priming of eASCs potentiated their antiproliferative and chondroprotective functions. We demonstrated that the immunocompetent mouse model of CIOA was relevant to test the therapeutic efficacy of xenogeneic eASCs for OA and confirmed that IFN&gamma;-primed eASCs may have a therapeutic value for musculoskeletal diseases in veterinary medicine.

No MeSH data available.