Belacic Zarah A, Sullivan Stasia N, Rice Hilary C, Durgam Sushmitha S
Am J Vet Res. 2023 Mar 19;84(4). doi: 10.2460/ajvr.22.08.0128. Print 2023 Apr 1.
To investigate the effects of interleukin-1β (IL-1β) and methylprednisolone acetate (MPA) on equine intrabursal deep digital flexor tendon (DDFT) and navicular bone fibrocartilage (NBF) cells in vitro.
Third passage DDFT and NBF cells from 5 healthy donor horses ages 11-17 years euthanized for reasons unrelated to musculoskeletal conditions.
Aggregate cultures were incubated with culture medium alone (control), 10 ng/mL IL-1β, 10 ng/mL IL-1β + 0.05 mg/mL MPA, or 10 ng/mL IL-1β + 0.5 mg/mL MPA for 24 hours. Extracellular matrix (ECM) gene expressions were assessed via real-time polymerase chain reaction (rtPCR). Culture media matrix metalloproteinase (MMP) -3 and -13 concentrations were quantified via ELISA. Total glycosaminoglycan (GAG) content in the cell pellets and culture media was also assessed.
IL-1β and IL-1β combined with MPA significantly downregulated ECM gene expression to a greater extent in NBF cells compared with DDFT cells. IL-1β and IL-1β combined with MPA significantly upregulated MMP-3 culture media concentrations in DDFT cells only, and MMP-13 culture media concentrations to a greater extent in NBF cells compared with DDFT cells.
NBF cells were more susceptible to IL-1β and MPA-mediated ECM gene expression downregulation in vitro. These results serve as a first step for future work to determine intrabursal corticosteroid regimens that limits or resolve the inflammation as well as take into consideration NBF cell biosynthesis in horses with navicular disease, for which currently no information exists.
研究白细胞介素-1β(IL-1β)和醋酸甲泼尼龙(MPA)对马掌内深屈肌腱(DDFT)和舟状骨纤维软骨(NBF)细胞的体外作用。
从5匹年龄为11 - 17岁、因与肌肉骨骼疾病无关的原因实施安乐死的健康供体马获取第三代DDFT和NBF细胞。
将聚集培养物分别与单纯培养基(对照)、10 ng/mL IL-1β、10 ng/mL IL-1β + 0.05 mg/mL MPA或10 ng/mL IL-1β + 0.5 mg/mL MPA孵育24小时。通过实时聚合酶链反应(rtPCR)评估细胞外基质(ECM)基因表达。通过酶联免疫吸附测定(ELISA)定量测定培养基中基质金属蛋白酶(MMP)-3和-13的浓度。还评估了细胞沉淀和培养基中总糖胺聚糖(GAG)的含量。
与DDFT细胞相比,IL-1β以及IL-1β与MPA联合使用在NBF细胞中更显著地下调ECM基因表达。IL-1β以及IL-1β与MPA联合使用仅在DDFT细胞中显著上调MMP-3培养基浓度,而与DDFT细胞相比,在NBF细胞中更显著地上调MMP-13培养基浓度。
在体外,NBF细胞对IL-1β和MPA介导的ECM基因表达下调更敏感。这些结果是未来工作的第一步,未来工作旨在确定限制或消除炎症的掌内皮质类固醇治疗方案,并考虑舟状骨疾病马匹的NBF细胞生物合成,目前尚无相关信息。