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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.


Related in: MedlinePlus

IFNγ-primed eASCs improve OA score. (A) OA score for tibia plateaus and femur condyles in the knee joint and mean at euthanasia. Results are expressed as the mean ± SEM, n = 10. (B) Osteoprotegerin (OPG) and (C) cartilage oligomeric matrix protein (COMP) concentrations were determined in sera of mice at euthanasia by specific sandwich enzyme-linked immunosorbent assay (ELISA). Results are expressed as the mean ± SEM, n = 10. (D) Proliferation of murine splenocytes in presence of different ratios of naïve or IFNγ-primed eASCs. Results are expressed as the percentage of ConA-induced proliferation of splenocytes which was assigned the value of 100% and represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Kruskal–Wallis test followed by Dunn’s test for multiple comparisons in A, B, and C and using ANOVA followed by a Dunnett’s test for multiple comparisons in D. *p < 0.05 or p = 0.07 compared samples with the control; #p < 0.05 compared samples with each other.
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Figure 4: IFNγ-primed eASCs improve OA score. (A) OA score for tibia plateaus and femur condyles in the knee joint and mean at euthanasia. Results are expressed as the mean ± SEM, n = 10. (B) Osteoprotegerin (OPG) and (C) cartilage oligomeric matrix protein (COMP) concentrations were determined in sera of mice at euthanasia by specific sandwich enzyme-linked immunosorbent assay (ELISA). Results are expressed as the mean ± SEM, n = 10. (D) Proliferation of murine splenocytes in presence of different ratios of naïve or IFNγ-primed eASCs. Results are expressed as the percentage of ConA-induced proliferation of splenocytes which was assigned the value of 100% and represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Kruskal–Wallis test followed by Dunn’s test for multiple comparisons in A, B, and C and using ANOVA followed by a Dunnett’s test for multiple comparisons in D. *p < 0.05 or p = 0.07 compared samples with the control; #p < 0.05 compared samples with each other.

Mentions: We, therefore, compared the effect of naïve and IFNγ-primed eASCs on OA progression. Injection of a high dose of eASCs (2 × 105 cells) significantly improved the OA score, while the low dose (2 × 104 eASCs) had no effect (Figure 4A). By contrast, both doses of IFNγ-primed eASCs reduced the OA score, but only the lowest dose of IFNγ-primed eASCs induced a significant decrease. Of importance, 2 × 104 IFNγ-primed eASCs were as efficient as 2 × 105 naïve eASCs to protect cartilage from degradation. The therapeutic effect of eASCs was confirmed by measuring the level of OPG, a bone metabolism marker, in the sera of mice at euthanasia. It was significantly decreased in mice treated with 2 × 104 IFNγ-primed eASCs (Figure 4B). Levels of COMP, a marker of cartilage turnover, did not change in the sera of treated mice compared with control group (Figure 4C), and IL6, IL10, IL1β, and C-terminal cross-linked telopeptide of type II collagen (CTX2) were not detected in the sera of mice (data not shown).


Utility of a Mouse Model of Osteoarthritis to Demonstrate Cartilage Protection by IFN γ -Primed Equine Mesenchymal Stem Cells
IFNγ-primed eASCs improve OA score. (A) OA score for tibia plateaus and femur condyles in the knee joint and mean at euthanasia. Results are expressed as the mean ± SEM, n = 10. (B) Osteoprotegerin (OPG) and (C) cartilage oligomeric matrix protein (COMP) concentrations were determined in sera of mice at euthanasia by specific sandwich enzyme-linked immunosorbent assay (ELISA). Results are expressed as the mean ± SEM, n = 10. (D) Proliferation of murine splenocytes in presence of different ratios of naïve or IFNγ-primed eASCs. Results are expressed as the percentage of ConA-induced proliferation of splenocytes which was assigned the value of 100% and represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Kruskal–Wallis test followed by Dunn’s test for multiple comparisons in A, B, and C and using ANOVA followed by a Dunnett’s test for multiple comparisons in D. *p < 0.05 or p = 0.07 compared samples with the control; #p < 0.05 compared samples with each other.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037129&req=5

Figure 4: IFNγ-primed eASCs improve OA score. (A) OA score for tibia plateaus and femur condyles in the knee joint and mean at euthanasia. Results are expressed as the mean ± SEM, n = 10. (B) Osteoprotegerin (OPG) and (C) cartilage oligomeric matrix protein (COMP) concentrations were determined in sera of mice at euthanasia by specific sandwich enzyme-linked immunosorbent assay (ELISA). Results are expressed as the mean ± SEM, n = 10. (D) Proliferation of murine splenocytes in presence of different ratios of naïve or IFNγ-primed eASCs. Results are expressed as the percentage of ConA-induced proliferation of splenocytes which was assigned the value of 100% and represented as mean ± SEM for three independent biological replicates. Data were analyzed using the Kruskal–Wallis test followed by Dunn’s test for multiple comparisons in A, B, and C and using ANOVA followed by a Dunnett’s test for multiple comparisons in D. *p < 0.05 or p = 0.07 compared samples with the control; #p < 0.05 compared samples with each other.
Mentions: We, therefore, compared the effect of naïve and IFNγ-primed eASCs on OA progression. Injection of a high dose of eASCs (2 × 105 cells) significantly improved the OA score, while the low dose (2 × 104 eASCs) had no effect (Figure 4A). By contrast, both doses of IFNγ-primed eASCs reduced the OA score, but only the lowest dose of IFNγ-primed eASCs induced a significant decrease. Of importance, 2 × 104 IFNγ-primed eASCs were as efficient as 2 × 105 naïve eASCs to protect cartilage from degradation. The therapeutic effect of eASCs was confirmed by measuring the level of OPG, a bone metabolism marker, in the sera of mice at euthanasia. It was significantly decreased in mice treated with 2 × 104 IFNγ-primed eASCs (Figure 4B). Levels of COMP, a marker of cartilage turnover, did not change in the sera of treated mice compared with control group (Figure 4C), and IL6, IL10, IL1β, and C-terminal cross-linked telopeptide of type II collagen (CTX2) were not detected in the sera of mice (data not shown).

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.


Related in: MedlinePlus