Duesterdieck-Zellmer Katja F, Driscoll Nellie, Ott Jesse F
Department of Clinical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
Am J Vet Res. 2012 Oct;73(10):1530-9. doi: 10.2460/ajvr.73.10.1530.
To determine concentration-dependent effects of tiludronate on cartilage explants incubated with or without recombinant equine interleukin-1β (rEq IL-1).
Articular cartilage explants from the femorotibial joints of 3 young adult horses.
Cartilage explants were incubated with 1 of 6 concentrations (0, 0.19, 1.9, 19, 190, or 1,900 mg/L) of tiludronate and with or without rEq IL-1 (0.01 ng/mL) for 96 hours. Prostaglandin E(2) (PGE(2)) concentrations in culture medium and explant digests were analyzed via PGE(2) enzyme immunoassay. Sulfated glycosaminoglycan (sGAG) concentrations in culture medium were quantified via 1,9-dimethylmethylene blue assay. Chondrocyte apoptosis in paraffin embedded explant sections was measured via terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay. Relative gene expression of matrix metalloproteinases (MMPs), interleukin (IL)-6, and IL-8 was determined via the comparative cycle threshold method.
rEq IL-1 increased PGE(2) concentration, sGAG release from explants, chondrocyte apoptosis, and MMP gene expression. Lower tiludronate concentrations reduced rEq IL-1-induced sGAG release and chondrocyte apoptosis, whereas the higher tiludronate concentrations increased sGAG release and chondrocyte apoptosis. At the highest tiludronate concentration evaluated, IL-8 gene expression was increased independent of whether rEq IL-1 was present.
Tiludronate had biphasic concentration-dependent effects on cartilage explants that were independent of PGE(2) secretion or MMP gene expression. Low tiludronate concentrations had some chondroprotective effects, whereas high tiludronate concentrations were detrimental to equine articular cartilage. Administration of tiludronate intra-articularly to horses may be detrimental, dependent on the dose used. In vivo studies are needed before intra-articular tiludronate administration to horses can be recommended.
确定替鲁膦酸盐对在添加或未添加重组马白细胞介素-1β（rEq IL-1）的情况下孵育的软骨外植体的浓度依赖性作用。
取自3匹成年青年马股胫关节的关节软骨外植体。
将软骨外植体与6种浓度（0、0.19、1.9、19、190或1900 mg/L）的替鲁膦酸盐之一孵育，并添加或不添加rEq IL-1（0.01 ng/mL），持续96小时。通过前列腺素E2（PGE2）酶免疫测定法分析培养基和外植体消化物中的PGE2浓度。通过1,9-二甲基亚甲蓝测定法定量培养基中的硫酸化糖胺聚糖（sGAG）浓度。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法测量石蜡包埋的外植体切片中的软骨细胞凋亡。通过比较循环阈值法确定基质金属蛋白酶（MMP）、白细胞介素（IL）-6和IL-8的相对基因表达。
rEq IL-1增加了PGE2浓度、外植体的sGAG释放、软骨细胞凋亡和MMP基因表达。较低浓度的替鲁膦酸盐降低了rEq IL-1诱导的sGAG释放和软骨细胞凋亡，而较高浓度的替鲁膦酸盐增加了sGAG释放和软骨细胞凋亡。在评估的最高替鲁膦酸盐浓度下，无论是否存在rEq IL-1，IL-8基因表达均增加。
替鲁膦酸盐对软骨外植体具有双相浓度依赖性作用，且与PGE2分泌或MMP基因表达无关。低浓度的替鲁膦酸盐具有一定的软骨保护作用，而高浓度的替鲁膦酸盐对马关节软骨有害。向马关节内注射替鲁膦酸盐可能有害，这取决于所用剂量。在推荐向马关节内注射替鲁膦酸盐之前，需要进行体内研究。
