Orthopaedic Research Unit, Department of Surgery, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, United Kingdom.
J Orthop Res. 2014 Jul;32(7):923-7. doi: 10.1002/jor.22622. Epub 2014 Apr 9.
The aim of this in vitro study was to ascertain the effect of recombinant human Fibroblast Growth Factor-18 (rhFGF18) on the repair response of mechanically damaged articular cartilage. Articular cartilage discs were harvested from healthy mature horses (n = 4) and subjected to single impact load (SIL). The impacted explants, together with unimpacted controls were cultured in modified DMEM ± 200 ng/ml rhFGF18 for up to 30 days. Glycosaminoglycan (GAG) release into the media was measured using the dimethylmethylene blue (DMMB) assay. Aggrecan neopepitope CS846, collagen type II synthesis (CPII) and cleavage (C2C) were measured by ELISA. Histological analysis and TUNEL staining were used to assess repair cell number and cell death. Impacted explants treated with rhFGF18 showed significantly more GAG and CS846 release into the media (p < 0.05), there was also a significant decrease in C2C levels at Day 20. Loaded sections treated with rhFGF18 had more repair cells and significantly less cell death (p < 0.001) at Day 30 in culture. In an in vitro damage/repair model, rhFGF18 increases the proteoglycan synthesis, the repair cell number and prevents apoptosis at Day 30. This suggests that rhFGF18 may be a good candidate for enhancement of cartilage repair following mechanical damage.
本体外研究旨在确定重组人成纤维细胞生长因子 18（rhFGF18）对机械损伤关节软骨修复反应的影响。从健康成熟马（n=4）中采集关节软骨盘，并进行单次冲击负荷（SIL）。受冲击的标本与未受冲击的对照标本一起在改良 DMEM+/-200ng/ml rhFGF18 中培养，最长可达 30 天。使用二甲亚甲基蓝（DMMB）测定法测量糖胺聚糖（GAG）释放到培养基中的情况。通过 ELISA 测量聚集蛋白聚糖 neo 表位 CS846、型胶原合成（CPII）和裂解（C2C）。组织学分析和 TUNEL 染色用于评估修复细胞数量和细胞死亡。用 rhFGF18 处理的受冲击标本显示出显著更多的 GAG 和 CS846 释放到培养基中（p<0.05），在第 20 天时 C2C 水平也显著降低。用 rhFGF18 处理的负载标本在培养第 30 天时有更多的修复细胞，且细胞死亡显著减少（p<0.001）。在体外损伤/修复模型中，rhFGF18 在第 30 天增加了蛋白聚糖合成、修复细胞数量并预防了细胞凋亡。这表明 rhFGF18 可能是机械损伤后增强软骨修复的良好候选物。
