Linardi Renata L, Dodson Michael E, Moss Kaitlyn L, King William J, Ortved Kyla F
Department of Clinical Studies, New Bolton Center, University of Pennsylvania, Kennett Square, PA, United States.
Owl Manor Veterinary Inc., Warsaw, IN, United States.
Front Vet Sci. 2019 Mar 6;6:64. doi: 10.3389/fvets.2019.00064. eCollection 2019.
Cartilage injury occurs commonly in equine athletes, often precipitating posttraumatic osteoarthritis (PTOA). Orthobiologics such as autologous conditioned serum (ACS) and autologous protein solution (APS) may be useful in decreasing posttraumatic inflammation, thereby preventing PTOA. The objective of this study was to quantify cytokine concentrations in ACS and APS and evaluate the protective effects of ACS and APS on inflamed chondrocytes cultured . We hypothesized that the combination of platelet-derived growth factors (PDGF) and anti-inflammatory cytokines present in APS would be superior in decreasing the inflammatory and catabolic cascade in inflamed chondrocytes when compared to ACS in which platelets are excluded from the preparation. Chondrocytes were isolated from the cartilage of femoral trochlear ridges of 6 horses and cultured in 12-well transwell plates. Treatment groups included: (1) control, (2) APS (Pro-Stride; Owl Manor), and (3) ACS (IRAP II; Arthrex). Each group was unstimulated or stimulated with IL-1β and TNF-α for 48 h. The concentration of IL-1β, IL-6, TNF-α, MMP-3, MMP-13, and IL-10 was quantified using a fluorescent bead-based multiplex assay. IL-1Ra concentration was quantified using ELISA. APS and ACS both had significantly increased concentrations of IL-1Ra without a concurrent increase in IL-1β concentration. After 48 h of culture, media from chondrocytes treated with APS contained significantly increased concentrations of IL-1Ra and IL-10. APS-treated cultures had increased concentrations of IL-6. Overall, APS effectively concentrated IL-1Ra without an incubation period and media from APS-treated chondrocytes had increased concentrations of chondroprotective (IL-1Ra and IL-10) and modulatory (IL-6) cytokines, which may be beneficial in the treatment of inflammatory conditions such as PTOA.
软骨损伤在马运动员中很常见,常引发创伤后骨关节炎(PTOA)。诸如自体条件血清(ACS)和自体蛋白溶液(APS)等骨科生物制剂可能有助于减轻创伤后炎症,从而预防PTOA。本研究的目的是量化ACS和APS中的细胞因子浓度,并评估ACS和APS对培养的炎症软骨细胞的保护作用。我们假设,与制备过程中不含血小板的ACS相比,APS中存在的血小板衍生生长因子(PDGF)和抗炎细胞因子的组合在减少炎症软骨细胞中的炎症和分解代谢级联反应方面更具优势。从6匹马的股骨滑车嵴软骨中分离软骨细胞,并在12孔Transwell板中培养。治疗组包括:(1)对照组,(2)APS(Pro-Stride;Owl Manor),和(3)ACS(IRAP II;Arthrex)。每组在未刺激或用IL-1β和TNF-α刺激48小时。使用基于荧光微球的多重检测法定量IL-1β、IL-6、TNF-α、MMP-3、MMP-13和IL-10的浓度。使用ELISA定量IL-1Ra浓度。APS和ACS的IL-1Ra浓度均显著增加,而IL-1β浓度没有同时增加。培养48小时后,用APS处理的软骨细胞培养基中IL-1Ra和IL-10的浓度显著增加。APS处理的培养物中IL-6浓度增加。总体而言,APS无需孵育期就能有效浓缩IL-1Ra,且来自APS处理的软骨细胞的培养基中具有软骨保护(IL-1Ra和IL-10)和调节(IL-6)细胞因子浓度增加,这可能对治疗诸如PTOA等炎症性疾病有益。