Quam Vivian G, Belacic Zarah A, Long Sidney, Rice Hilary C, Dhar Madhu S, Durgam Sushmitha
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.
Ballarat Veterinary Practice Equine Clinic, Miners Rest, Victoria, Australia.
Equine Vet J. 2025 Jan;57(1):232-242. doi: 10.1111/evj.14090. Epub 2024 Apr 8.
Safe, efficacious therapy for treating degenerate deep digital flexor tendon (DDFT) and navicular bone fibrocartilage (NBF) in navicular horses is critically necessary. While archetypal orthobiologic therapies for navicular disease are used empirically, their safety and efficacy are unknown. Mesenchymal stem cell-derived extracellular vesicles (EV) may overcome several limitations of current orthobiologic therapies.
To (1) characterise cytokine and growth factor profiles of equine bone marrow mesenchymal stem cell (BM-MSC)-derived extracellular vesicles (BM-EV) and (2) evaluate the in vitro anti-inflammatory and extracellular matrix (ECM) protective potentials of BM-EV on DDFT and NBF explant co-cultures in an IL-1β inflammatory environment.
In vitro experimental study.
Cytokines (IL-1β, IL-6, IL-10, IL-1ra and TNF-α) and growth factors (TGFβ1, VEGF, IGF1 and PDGF) in equine BM-EV isolated via ultracentrifugation and precipitation methods were profiled. Forelimb DDFT and NBF explant co-cultures from seven horses were exposed to media alone, or media containing 2 × 10 ± 0.1 × 10 particles/mL or 10 μg/mL BM-EV (BM-EV), 10 ng/mL interleukin-1β (IL-1β), or IL-1β + BM-EV for 48 h. Co-culture media IL-6, TNF-α, MMP-3, MMP-13 concentrations and explant sulphated glycosaminoglycan (sGAG) content were quantified.
IL-6, IGF1 and VEGF concentrations were 102.1 (37.61-256.2) and 182.3 (163.1-226.3), 72.3 (8-175.6) and 2.4 (0.1-2.6), 108.3 (38.3-709.1) and 211.4 (189.1-318.2) pg/mL per 2 × 10 ± 0.1 × 10 particles/mL or 10 μg/mL 10 μg of BM-EV isolated via ultracentrifugation and precipitation methods, respectively. Co-culture media MMP-3 in BM-EV- (p = 0.03) and BM-EV + IL-1β-treated (p = 0.01) groups were significantly lower than the respective media and IL-1β groups. DDFT explant sGAG content of BM-EV (p = 0.003) and BM-EV + IL-1β groups were significantly higher compared with IL-1β group.
Specimen numbers are limited, in vitro model may not replicate clinical case conditions, lack of non-MSC-derived EV control group.
Equine BM-EV contains IL-6 and growth factors, IGF1 and VEGF. The anti-inflammatory and ECM protective potentials of BM-EV were evident as increased IL-6 and decreased MMP-3 concentrations in the DDFT-NBF explant co-culture media. These results support further evaluation of BM-EV as an acellular and 'off-the-shelf' intra-bursal/intrasynovial therapy for navicular pathologies.
为患有舟状骨病的马匹治疗退化的指深屈肌腱（DDFT）和舟状骨纤维软骨（NBF），安全、有效的疗法至关重要。虽然治疗舟状骨病的典型的骨科生物疗法在经验上被使用，但其安全性和有效性尚不清楚。间充质干细胞衍生的细胞外囊泡（EV）可能克服当前骨科生物疗法的几个局限性。
（1）表征马骨髓间充质干细胞（BM-MSC）衍生的细胞外囊泡（BM-EV）的细胞因子和生长因子谱，（2）评估在白细胞介素-1β（IL-1β）炎症环境中，BM-EV对DDFT和NBF外植体共培养物的体外抗炎和细胞外基质（ECM）保护潜力。
体外实验研究。
对通过超速离心和沉淀法分离的马BM-EV中的细胞因子（IL-1β、IL-6、IL-10、IL-1ra和TNF-α）和生长因子（TGFβ1、VEGF、IGF1和PDGF）进行分析。将来自7匹马的前肢DDFT和NBF外植体共培养物分别暴露于单独的培养基、含有2×10±0.1×10颗粒/毫升或10μg/毫升BM-EV（BM-EV）的培养基、10ng/毫升白细胞介素-1β（IL-1β）或IL-1β+BM-EV中48小时。对共培养培养基中的IL-6、TNF-α、MMP-3、MMP-13浓度以及外植体硫酸化糖胺聚糖（sGAG）含量进行定量。
通过超速离心和沉淀法分离的每2×10±0.1×10颗粒/毫升或10μg/毫升10μg的BM-EV中，IL-6、IGF1和VEGF浓度分别为102.1（37.61-256.2）和182.3（163.1-226.3）、72.3（8-175.6）和2.4（0.1-2.6）、108.3（38.3-709.1）和211.4（189.1-318.2）pg/毫升。BM-EV组（p = 0.03）和BM-EV+IL-1β处理组（p = 0.01）共培养培养基中的MMP-3显著低于各自的培养基组和IL-1β组。与IL-1β组相比，BM-EV组（p = 0.003）和BM-EV+IL-1β组DDFT外植体的sGAG含量显著更高。
样本数量有限，体外模型可能无法复制临床病例情况，缺乏非MSC衍生的EV对照组。
马BM-EV含有IL-6以及生长因子IGF1和VEGF。在DDFT-NBF外植体共培养培养基中，BM-EV的抗炎和ECM保护潜力表现为IL-6增加和MMP-3浓度降低。这些结果支持进一步评估BM-EV作为一种用于舟状骨病变的无细胞和“现成可用”的囊内/滑膜内疗法。
