Seifer D R, Furman B D, Guilak F, Olson S A, Brooks S Carroll, Kraus V Byers
Department of Medicine, Division of Rheumatology, Duke University Medical Center, Durham, NC 27710, United States.
Osteoarthritis Cartilage. 2008 Dec;16(12):1532-8. doi: 10.1016/j.joca.2008.04.013. Epub 2008 Jun 5.
We evaluated three methodologies--a calcium sodium alginate compound (CSAC), polyacrylate beads (PABs), and Whatman paper recovery (WPR)--for the ability to recover synovial fluid (SF) from mouse knees in a manner that facilitated biochemical marker analysis.
Pilot testing of each of these recovery vehicles was conducted using small volumes of waste human SF. CSAC emerged as the method of choice, and was used to recover and quantify SF from the knees of C57BL/6 mice (n=12), six of which were given left knee articular fractures. SF concentrations of cartilage oligomeric matrix protein (COMP) were measured by enzyme-linked immunosorbent assay.
The mean concentration ratio [(COMP(left knee))/(COMP(right knee))] was higher in the mice subjected to articular fracture when compared to the non-fracture mice (P=0.026). The mean total COMP ratio (taking into account the quantitative recovery of SF) best discriminated between fracture and non-fracture knees (P=0.004).
Our results provide the first direct evidence of accelerated joint tissue turnover in a mouse model responding to acute joint injury. These data strongly suggest that mouse SF recovery is feasible and that biomarker analysis of collected SF samples can augment traditional histological analyses in mouse models of arthritis.
我们评估了三种方法——海藻酸钠钙复合物(CSAC)、聚丙烯酸酯珠(PABs)和沃特曼滤纸回收法(WPR)——从小鼠膝关节中回收滑液(SF)并便于进行生化标志物分析的能力。
使用少量废弃人体滑液对每种回收载体进行预试验。CSAC成为首选方法,并用于从C57BL/6小鼠(n = 12)的膝关节中回收和定量滑液,其中6只小鼠左膝关节发生关节骨折。通过酶联免疫吸附测定法测量软骨寡聚基质蛋白(COMP)的滑液浓度。
与未骨折小鼠相比,发生关节骨折的小鼠的平均浓度比[(COMP(左膝))/(COMP(右膝))]更高(P = 0.026)。平均总COMP比(考虑到滑液的定量回收)在骨折和未骨折膝关节之间的区分效果最佳(P = 0.004)。
我们的结果首次提供了在对急性关节损伤作出反应的小鼠模型中关节组织周转加速的直接证据。这些数据强烈表明,从小鼠回收滑液是可行的,并且对收集的滑液样本进行生物标志物分析可以增强关节炎小鼠模型中的传统组织学分析。