Distinct differences in immunological properties of equine orthobiologics revealed by functional and transcriptomic analysis using an activated macrophage readout system. Distinct differences in immunological properties of equine orthobiologics revealed by functional and transcriptomic analysis using an activated macrophage readout system

Multiple biological therapies for orthopedic injuries are marketed to veterinarians, despite a lack of rigorous comparative biological activity data to guide informed decisions in selecting a most effective compound. Therefore, the goal of this study was to use relevant bioassay systems to directly compare the anti-inflammatory and immunomodulatory activity of three commonly used orthobiological therapies (OTs): mesenchymal stromal cells (MSC), autologous conditioned serum (ACS), and platelet rich plasma (PRP). Equine monocyte-derived macrophages were used as the readout system to compare therapies, including cytokine production and transcriptomic responses. Overall design: Macrophages were stimulated with IL-1ß and treated 24 hours with orthobiologicss, washed and cultured an additional 24 hours to generate supernatants. Secreted cytokines were measured by multiplex immunoassay and ELISA. To assess global transcriptomic responses to treatments, RNA was extracted from macrophages and subjected to full RNA sequencing

目前市面上已有多款针对骨科损伤的生物疗法面向兽医推广，但目前缺乏严谨的比较生物学活性数据，以指导兽医在遴选最优有效制剂时做出明智决策。因此本研究旨在利用相关生物检测系统，直接对比三种常用骨科生物疗法（orthobiologic therapies, OTs）的抗炎与免疫调节活性：间充质基质细胞（mesenchymal stromal cells, MSC）、自体条件血清（autologous conditioned serum, ACS）以及富血小板血浆（platelet rich plasma, PRP）。本研究采用马源单核细胞衍生巨噬细胞作为检测体系，评估不同疗法的细胞因子产生情况与转录组响应。实验整体设计如下：将巨噬细胞经白细胞介素1β（IL-1β）刺激后，使用骨科生物疗法处理24小时，随后洗涤细胞并继续培养24小时以收集上清液。通过多重免疫分析法与酶联免疫吸附试验（ELISA）检测上清液中分泌的细胞因子水平。为评估各处理对细胞的全局转录组响应，我们从巨噬细胞中提取RNA并开展全转录组RNA测序。