Duan Xin, Sandell Linda J, Chinzei Nobuaki, Holguin Nilsson, Silva Matthew J, Schiavinato Antonella, Rai Muhammad Farooq
Department of Orthopaedic Surgery, Musculoskeletal Research Center, Washington University School of Medicine at Barnes-Jewish Hospital, St. Louis, Missouri, United States of America.
Department of Orthopedic Surgery, First Affiliated Hospital of Sun Yet-san University, Guangzhou, China.
PLoS One. 2017 Apr 13;12(4):e0175682. doi: 10.1371/journal.pone.0175682. eCollection 2017.
To investigate the therapeutic potential of intra-articular hyaluronan-derivative HYADD® 4-G and/or platelet-rich plasma (PRP) in a mouse model of non-invasive joint injury.
Non-invasive axial tibial loading was used to induce joint injury in 10-week-old C57BL/6J mice (n = 86). Mice underwent a single loading of either 6 Newton (N) or 9N axial tibial compression. HYADD® 4-G was injected intra-articularly at 8 mg/mL or 15 mg/mL either before or after loading with or without PRP. Phosphate-buffered-saline was injected as control. Knee joints were harvested at 5 or 56 days post-loading and prepared for micro-computed tomography scanning and subsequently processed for histology. Immunostaining was performed for aggrecan to monitor its distribution, for CD44 to monitor chondrocyte reactive changes and for COMP (cartilage oligomeric matrix protein) as an index for cartilage matrix changes related to loading and cartilage injury. TUNEL assay was performed to identify chondrocyte apoptosis.
Loading initiated cartilage proteoglycan loss and chondrocyte apoptosis within 5 days with slowly progressive post-traumatic osteoarthritis (no cartilage degeneration, but increased synovitis and ectopic calcification after 9N loading) at 56 days. Mice treated with repeated HYADD® 4-G (15 mg/mL) or HYADD® 4-G (8 mg/mL) ± PRP or PRP alone exhibited no significant improvement in the short-term (5 days) and long-term (56 days) consequences of joint loading except for a trend for improved bone changes compared to non-loaded joints.
While we failed to show an overall effect of intra-articular delivery of hyaluronan-derivative and/or PRP in reversing/protecting the pathological events in cartilage and synovium following joint injury, some bone alterations were relatively less severe with hyaluronan-derivative at higher concentration or in association with PRP.
在非侵入性关节损伤小鼠模型中研究关节内注射透明质酸衍生物HYADD® 4-G和/或富血小板血浆（PRP）的治疗潜力。
采用非侵入性轴向胫骨加载法诱导10周龄C57BL/6J小鼠（n = 86）发生关节损伤。小鼠接受6牛顿（N）或9N的单次轴向胫骨压缩加载。在加载前或加载后，联合或不联合PRP，以8 mg/mL或15 mg/mL的浓度关节内注射HYADD® 4-G。注射磷酸盐缓冲盐水作为对照。在加载后5天或56天采集膝关节，进行微型计算机断层扫描，随后进行组织学处理。进行聚集蛋白聚糖免疫染色以监测其分布，进行CD44免疫染色以监测软骨细胞反应性变化，进行软骨寡聚基质蛋白（COMP）免疫染色作为与加载和软骨损伤相关的软骨基质变化指标。进行TUNEL检测以鉴定软骨细胞凋亡。
加载后5天内引发软骨蛋白聚糖丢失和软骨细胞凋亡，56天时出现缓慢进展的创伤后骨关节炎（9N加载后无软骨退变，但滑膜炎和异位钙化增加）。重复注射HYADD® 4-G（15 mg/mL）或HYADD® 4-G（8 mg/mL）±PRP或单独注射PRP治疗的小鼠，除与未加载关节相比骨变化有改善趋势外，在关节加载的短期（5天）和长期（56天）后果方面均未显示出显著改善。
虽然我们未能证明关节内注射透明质酸衍生物和/或PRP在逆转/保护关节损伤后软骨和滑膜的病理事件方面的总体效果，但较高浓度的透明质酸衍生物或与PRP联合使用时，一些骨改变相对较轻。
