艾灸疗法对实验性骨关节炎大鼠关节软骨的保护作用

通过观察艾灸足三里、肾俞穴对实验性骨关节炎大鼠关节软骨中基质金属蛋白酶-1（MMP-1）和Ⅱ型胶原（collagen-Ⅱ）水平表达的影响,探讨艾灸足三里、肾俞穴治疗骨性关节炎（OA）在软骨代谢方面的作用机制。将32只SPF级健康雌性SD大鼠随机分为假手术组、模型组、西药组和艾灸组。通过切除双侧卵巢、切断右膝内侧副韧带及交叉韧带建立OA动物模型。术后第4周,艾灸组给予艾灸足三里和肾俞穴治疗,西药组灌服西乐葆,假手术组与模型组灌服等量的生理盐水,于术后第8周处死动物,采用免疫组织化法测定各组关节软骨中MMP-1及Ⅱ型胶原的表达情况,对比各组数据。结果术后第8周假手术组中MMP-1的表达量很少;MMP-1在模型组、西药组、艾灸组中表达均较假手术组明显增多（P〈0.01）;艾灸组中MMP-1表达与模型组与西药组比较均明显减少（P〈0.01）。关节软骨中Ⅱ型胶原的表达量在模型组、西药组、艾灸组中均较假手术组明显减少（P〈0.01）;艾灸组中Ⅱ型胶原的表达量与模型组、西药组比较均明显增多（P〈0.01,P〈0.05）。说明艾灸足三里、肾俞穴能够提高OA软骨的自身修复能力,其软骨保护作用可能是通过降低关节软骨中MMP-1的表达与减缓Ⅱ型胶原的降解速度来实现的。     

艾灸疗法对实验性骨关节炎大鼠关节软骨的保护作用

通过观察艾灸足三里、肾俞穴对实验性骨关节炎大鼠关节软骨中基质金属蛋白酶-1(MMP-1)和Ⅱ型胶原(collagen-Ⅱ)水平表达的影响,探讨艾灸足三里、肾俞穴治疗骨性关节炎(OA)在软骨代谢方面的作用机制。将32只SPF级健康雌性SD大鼠随机分为假手术组、模型组、西药组和艾灸组。通过切除双侧卵巢、切断右膝内侧副韧带及交叉韧带建立OA动物模型。术后第4周,艾灸组给予艾灸足三里和肾俞穴治疗,西药组灌服西乐葆,假手术组与模型组灌服等量的生理盐水,于术后第8周处死动物,采用免疫组织化法测定各组关节软骨中MMP-1及Ⅱ型胶原的表达情况,对比各组数据。结果术后第8周假手术组中MMP-1的表达量很少;MMP-1在模型组、西药组、艾灸组中表达均较假手术组明显增多(P0.01);艾灸组中MMP-1表达与模型组与西药组比较均明显减少(P0.01)。关节软骨中Ⅱ型胶原的表达量在模型组、西药组、艾灸组中均较假手术组明显减少(P0.01);艾灸组中Ⅱ型胶原的表达量与模型组、西药组比较均明显增多(P0.01,P0.05)。说明艾灸足三里、肾俞穴能够提高OA软骨的自身修复能力,其软骨保护作用可能是通过降低关节软骨中MMP-1的表达与减缓Ⅱ型胶原的降解速度来实现的。     

SD大鼠骨关节炎造模方法的筛选及优化探究

通过几种具有代表性的造模方法诱导膝关节骨性关节炎动物模型,筛选出理想的骨性关节炎造模方法。将32只SPF级健康雌性SD大鼠随机分为空白组、手术组、关节腔注射1组、关节腔注射2组,每组8只。手术组:Hulth造模法复制膝骨性关节炎动物模型,切断右侧膝交叉及内侧副韧带;关节腔注射1组:右侧注射4%木瓜蛋白酶溶液0.3 mL;关节腔注射2组:右侧注射1 g/L碘乙酸钠溶液0.1 mL。于术后第4周抽取关节液作一氧化氮(NO)和前列腺素E2(PGE2)测定;处死动物,采用骨性关节炎软骨病理变化评价系统(OARSI)评价关节软骨的病变;采用光镜观察软骨细胞生长情况;运用免疫组化法测定关节软骨中基质金属蛋白酶-1(MMP-1)的含量,对比各组数据。术后第4周,手术组病理改变OARSI评分、软骨细胞的MMP-1阳性表达和关节液中的NO、PGE2,与关节腔注射组相比均有极显著性差异(P<0.01);关节腔注射1组与关节腔注射2组相比,各项指标水平无显著性差异(P>0.05)。试验结果提示:手术组Hulth造模法复制膝骨性关节炎动物模型(通过切断右侧膝交叉及内侧副韧带建立骨性关节炎动物模型)是建立膝关节骨性关节炎动物模型的理想方法。     

双合汤对膝骨性关节炎大鼠关节液中PGE_2、MDA、SOD及MMP-3的影响

探讨双合汤对膝骨性关节炎大鼠关节液PGE_2、MDA、SOD、MMP-3的影响。通过膝关节腔注射木瓜蛋白酶法建立膝骨性关节炎大鼠模型(2周),将57只造模成功大鼠随机分为模型组(2mL/kg)、氨基葡萄糖组(86.4 mg/kg)和双合汤组(12g/kg),20只膝关节腔注射等量生理盐水大鼠为假手术组(2mL/kg)。共用相应药物灌胃4周。检测各组大鼠2周和6周关节活动范围、关节滑膜厚度和关节液中前列腺素(PG)E_2、丙二醛(MDA)、超氧化物歧化酶(SOD)、基质金属蛋白酶3(MMP-3)的水平。结果表明,与假手术组比较,模型组关节活动范围减小(P0.05),关节滑膜厚度增加(P0.05),关节液PGE_2、MDA、MMP-3水平显著升高(P0.05),SOD水平显著减少(P0.05);与模型组比较,双合汤组关节活动范围增加(P0.05),关节滑膜厚度减少(P0.05),关节液PGE_2、MDA、MMP-3水平显著降低(P0.05),SOD水平显著升高(P0.05)。表明双合汤能有效治疗膝骨性关节炎,其机制可能与抑制关节液中PGE_2、MDA、MMP-3的表达和促进SOD的表达有关。     

前列腺素E2和F2α对LPS刺激后奶牛子宫内膜上皮细胞COX-1和COX-2表达的影响

试验旨在阐明前列腺素E₂（prostaglandin E₂,PGE₂）和F_2α（prostaglandin F_2α,PGF_2α）对体外培养的奶牛子宫内膜上皮细胞中环氧合酶-1（cyclooxygenase-1,COX-1））与环氧合酶-2（cyclooxygenase-2,COX-2）表达的影响。培养奶牛子宫内膜上皮原代细胞和传代细胞,第4代细胞以1×10⁶个/孔接种于6孔板,以10^-7mol/L PGE₂和PGF_2α分别预处理细胞24 h,以100 ng/mL细菌脂多糖（lipopolysaccharides,LPS）刺激细胞4、8和12 h后分别提取RNA和总蛋白质,采用实时荧光定量PCR与Western blotting等技术检测COX-1与COX-2 mRNA和蛋白质的表达量。结果表明,与对照组相比,COX-1 mRNA表达量在PGE₂单独作用4、8和12 h后显著上调（P<0.05）;COX-2 mRNA表达量在PGE₂单独作用4和12 h后显著上调（P<0.05）,PGE₂单独处理使COX-1、COX-2蛋白表达量均显著上调（P<0.05）。与对照组相比,LPS刺激8和12 h时COX-1 mRNA表达量显著下调（P<0.05）,LPS刺激后COX-1蛋白表达量无显著变化（P>0.05）;LPS刺激后4、8和12 h时COX-2 mRNA表达量显著上调（P<0.05）,LPS刺激后COX-2蛋白表达量显著上调（P<0.05）。与LPS单独处理组相比,LPS+PGE₂处理组在8和12 h时COX-1和COX-2 mRNA表达量均显著上调（P<0.05）,同时COX-1和COX-2蛋白表达量也显著上调（P<0.05）。PGF_2α在LPS未刺激和刺激后对COX-1和COX-2 mRNA的表达无显著影响（P>0.05）,仅在PGF_2α单独处理8和12 h后COX-1 mRNA表达量上调（P<0.05）。两种激素联合处理与各自单独处理及LPS单独刺激相比,对COX-1和COX-2 mRNA表达具有一定的协同诱导作用。     

双合汤对膝骨性关节炎大鼠作用的试验

为了研究双合汤对大鼠膝骨性关节炎的作用及可能机制,通过膝关节腔注射木瓜蛋白酶法建立膝骨性关节炎大鼠模型(2周),将57只造模成功大鼠随机分为模型组(2 m L/kg·d)、氨基葡萄糖组(86.4 mg/kg·d)和双合汤组(12 g/kg·d),20只膝关节腔注射等量生理盐水大鼠为假手术组(2 m L/kg·d)。共用相应药物灌胃4周。检测各组大鼠2周和6周关节活动范围、关节滑膜厚度和关节液中IL-1β、IL-6、NO、一氧化氮合酶(i NOS)的水平。结果表明,与假手术组比较,模型组关节活动范围减少(P0.05),关节滑膜厚度增加(P0.05),关节液IL-1β、IL-6、NO、i NOS水平显著升高(P0.05);与模型组比较,双合汤组关节活动范围增加(P0.05),关节滑膜厚度减少(P0.05),关节液IL-1β、IL-6、NO、i NOS水平显著降低(P0.05)。说明双合汤能有效治疗膝骨性关节炎,其机制可能与抑制关节液中IL-1β、IL-6、NO、i NOS的表达有关。     

有氧运动对兔骨关节炎软骨细胞凋亡及相关基因蛋白表达的影响

探讨有氧运动对兔骨关节炎软骨细胞凋亡及Bcl-2、Bax、MMP-2、MMP-13、β-catenin、GSK-3β蛋白表达的影响。建立兔骨关节炎(OA)模型,试验分为空白对照组、模型对照组、低强度运动组、中等强度运动组。TUNEL标记法检测不同强度有氧运动对关节炎软骨细胞凋亡的影响;Western blot检测Bcl-2、Bax、MMP-2、MMP-13、β-catenin、GSK-3β蛋白表达。模型对照组、低强度运动组、中等强度运动组关节炎软骨细胞凋亡率均显著高于空白对照组,低强度运动组、中等强度运动组关节炎软骨细胞凋亡率均显著低于模型对照组,中等强度运动组关节炎软骨细胞凋亡率显著低于低强度运动组(P0.01);模型对照组、低强度运动组、中等强度运动组关节炎软骨细胞Bcl-2、MMP-2、MMP-13、β-catenin、GSK-3β蛋白表达均显著高于空白对照组,Bax蛋白表达显著低于空白对照组;低强度运动组、中等强度运动组关节炎软骨细胞Bcl-2、MMP-2、MMP-13、β-catenin、GSK-3β蛋白表达均显著低于模型组对照组,Bax蛋白表达显著高于模型对照组;中等强度运动组关节炎软骨细胞Bcl-2、MMP-2、MMP-13、β-catenin、GSK-3β蛋白表达均显著低于低强度运动组,Bax蛋白表达显著高于低强度运动组(P0.01)。不同强度有氧通过调节Bcl-2和Bax蛋白表达降低关节炎软骨细胞凋亡,并下调MMP-2、MMP-13,降低软骨细胞的损伤,其机制与下调Wnt/β-catenin信号通路有关。     

艾灸足三里、肾俞穴对骨性关节炎模型大鼠PGE2、NO 及 IL-1β的影响

观察艾灸足三里、肾俞穴对骨性关节炎（OA）模型大鼠关节液中前列腺素E2（PGE2）、一氧化氮（NO）及血清中白细胞介素-1β（IL-1β）水平的影响，进一步研究艾灸足三里、肾俞穴干预 OA 炎症发生发展的作用机制。将24只 SPF 级健康雌性 SD 大鼠随机分为 Sham 组、对照组和艾灸组。通过切除双侧卵巢并切断右侧膝交叉及内侧副韧带建立 OA 动物模型，术后第4周，艾灸组给予艾灸足三里和肾俞穴治疗，Sham组及模型灌服2 ml 的生理盐水，分别于术后第4、8周采用酶联免疫吸附法（ELISA）法测定关节液中 PGE2、NO 及血清中 IL-1β的水平，对比各组数据。术后第 8周，艾灸组关节液中的 PGE2、NO 较 Sham 组明显升高（P ＜0．01），较对照组明显减少（P ＜0．05），较 Sham 组明显升高（P ＜0．01），艾灸组血清中 IL-1β的水平较对照组明显减弱（P ＜0．01）；艾灸组关节液中的 PGE2较术后第4周明显减少（P ＜0．05），其关节液中的 NO 及血清中 IL-1β较术后第4周明显减少（P ＜0．01）。艾灸足三里、肾俞穴能明显抑制 OA 大鼠关节液中 PGE2、NO 及血清中的 IL-1β水平，减缓 OA 炎症的发生发展，可作为 OA 的有效治疗方法。     

烟碱对LPS诱导的兔子宫内膜上皮细胞炎症因子及前列腺素分泌的影响

试验旨在探索α7烟碱乙酰胆碱受体（α7 nicotinic acetylcholine receptor,α7nAChR）的高亲和力激动剂烟碱对脂多糖（lipopolysaccharides,LPS）诱导的兔子宫内膜上皮炎症的作用机制。分离发情后期兔的子宫内膜上皮细胞,用100 ng/mL LPS对细胞进行炎性刺激12 h。用CCK8法检测不同浓度烟碱（5、10和20 μg/mL）对细胞存活率的影响,筛选合适浓度的烟碱进行后续试验。将细胞分为对照组（CON）、LPS、LPS+烟碱、LPS+烟碱+甲基牛扁碱（MLA）（α7nAChR的特异性颉颃剂）组,通过ELISA法检测细胞培养上清液中白细胞介素-1β（interleukin 1β,IL-1β）、IL-6、IL-8、肿瘤坏死因子-α（tumor necrosis factor,TNF-α）、前列腺素E₂（prostaglandin E₂,PGE₂）和前列腺素F_2α（PGF_2α）的含量。试验成功分离兔子宫内膜上皮细胞,且传至第5代仍保持良好的生长状态。CCK8检测结果显示,20 μg/mL烟碱组细胞存活率显著降低（P<0.05）,10 μg/mL烟碱对细胞存活率无显著影响（P>0.05）,所以选择10 μg/mL烟碱进行后续试验。ELISA结果显示,与对照组相比,LPS组IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α的含量显著增加（P<0.05）;与LPS组相比,LPS+烟碱组显著降低IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α的含量（P<0.05）,LPS+烟碱+MLA组炎性因子和前列腺素的含量差异不显著（P<0.05）。以上结果表明,烟碱对LPS诱导的兔子宫内膜上皮细胞分泌IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α具有下调作用,推测烟碱通过α7nAChR介导炎性因子和前列腺素分泌下调而发挥抗炎作用,该结果可为研究α7nAChR作为子宫内膜炎治疗靶点的药物选择提供参考。     

螺旋藻多糖与银杏叶提取物复方对四氧嘧啶模型小鼠的降血糖作用

为研究复合螺旋藻多糖与银杏有效成分的降血糖作用,试验选用SPF级昆明种雄性小鼠,腹腔注射0.2 mL 200 mg/（kg·d）四氧嘧啶,连续5 d,断尾采血测定空腹血糖,其值大于11.1 mmol/L时表明造模成功。试验小鼠随机分为空白对照组（C组）、模型组（M组）、阳性对照组（CY组）、单一用药组（螺旋藻多糖组（P组）、银杏黄酮组（F组）、银杏内酯组（L组）,复合用药组将螺旋藻多糖（PSP）与银杏叶有效成分（GBE）配制成1：1（CP₁组）、2：1（CP₂组）、1：2（CP₃组）的复合组,每组10只,共9组。对照和CY组每只小鼠灌服2.5 mg/（kg·d）格列本脲,其余各组按200 mg/（kg·d）灌服相应药物,每天1次,连续灌服30 d,测定小鼠血糖值及体重、脾脏指数、胸腺指数及肝糖原等指标。结果显示,与M组相比,用药组小鼠体重均极显著增加（P<0.01）,复合用药组药效高于单一用药组;各用药组脾脏、胸腺指数均极显著升高（P<0.01）;肝糖原含量极显著增多（P<0.01）;血糖值极显著下降（P<0.01）;其中CP₂组的小鼠血糖值降幅最大,其降糖率为52.14%;肝糖原增加为61.88%。综上所述,螺旋藻多糖和银杏叶有效成分发生了协同增效作用,对由四氧嘧啶引起的高血糖小鼠有明显的降血糖作用。     

Comparison of the effects of caudal pole hemi-meniscectomy and complete medial meniscectomy in the canine stifle joint

OBJECTIVE: To compare the effects of caudal pole hemi-meniscectomy (CPHM) and complete medial meniscectomy (MM), specifically with respect to development of secondary osteoarthritis, in the stifle joints of clinically normal dogs. ANIMALS: 14 large-breed dogs. PROCEDURE: Unilateral CPHM (7 dogs) or MM (7) was performed, and the left stifle joints served as untreated control joints. Gait was assessed in all dogs before surgery and at 4, 8, 12, and 16 weeks postoperatively. After euthanasia, joints were evaluated grossly; Mankin cartilage scores, subchondral bone density assessment, and articular cartilage proteoglycan extraction and western blot analyses of 3B3(-) and 7D4 epitopes were performed. RESULTS: Weight distribution on control limbs exceeded that of treated limbs at 4 and 16 weeks after surgery in the CPHM group and at 4 and 8 weeks after surgery in the MM group; weight distribution was not significantly different between the 2 groups. After 16 weeks, incomplete meniscal regeneration and cartilage fibrillation on the medial aspect of the tibial plateau and medial femoral condyle were detected in treated joints in both groups. Mankin cartilage scores, subchondral bone density, and immunoexpression of 3B3(-) or 7D4 in articular cartilage in CPHM- or MM-treated joints were similar; 7D4 epitope concentration in synovial fluid was significantly greater in the MM-treated joints than in CPHM-treated joints. CONCLUSIONS AND CLINICAL RELEVANCE: Overall severity of secondary osteoarthritis induced by CPHM and MM was similar. Investigation of 7D4 epitope concentration in synovial fluid suggested that CPHM was associated with less disruption of chondrocyte metabolism.     

Recombinant equine growth hormone administration: effects on synovial fluid biomarkers and cartilage metabolism in horses

REASONS FOR PERFORMING STUDY: Recombinant equine growth hormone (reGH) has recently been evaluated for effects on body condition and wound healing. It has the potential to influence articular cartilage via stimulation of IGF-1. OBJECTIVES: To investigate effects of administration on synovial joint metabolism. METHODS: Six mature horses were given 20 microg/kg bwt reGH daily for 8 weeks by i.m. injection. Three control horses were injected with sterile water. Serum and synovial fluid samples were collected at 6, 8, 11 and 16 weeks for GH and IGF-1 assays. Articular cartilage harvested at week 16 was evaluated by Western analysis using monoclonal antibodies BC-13, BC-4, 8-A-4 and CH-3. RESULTS: Concentrations of IGF-1 in serum and synovial fluid were significantly elevated (P < 0.05) at 6 and 8 weeks in the reGH group. Glycosaminoglycan concentrations in synovial fluid were significantly less than controls at these time points, suggesting that reGH may modulate proteoglycan metabolism in articular cartilage. In the reGH group, there were not any alterations in synovial fluid content of 3B3(-) epitope or aggrecan metabolite, or in aggrecan or link protein catabolites retained within cartilage, that might be expected with development of osteoarthritis. CONCLUSIONS: Intramuscular administration of reGH may be a more efficient means of delivery of IGF-1 to joints for cartilage resurfacing initiatives. POTENTIAL RELEVANCE: We found no alterations in cartilage metabolism indicative of development of osteoarthritis.     

Metalloproteinases and tumor necrosis factor-alpha activities in synovial fluids of horses: correlation with articular cartilage alterations

Early detection of osteoarthritis in horses represents a challenge for equine practitioners. Several biological markers have been implicated in the pathological processes involved in articular cartilage destruction. To further document cartilage matrix proteases production, synovial fluid was collected from 14 horses (90 joints) before they were subjected to euthanasia. Growth macroscopic examination of the joints gave information on cartilage alterations. Samples were analyzed for matrix metalloproteinase (MMPs) activities by gelatin zymography and tumor necrosis factor alpha (TNF-alpha) cytotoxicity using L929 cells. Significant increase of MMP-9 monomer and dimer were found in synovial fluids of joints with severe cartilage alterations. On the contrary, the activity of TNF-alpha was not correlated to the degree of joint damage. The levels of MMP-9 monomer and dimer in the synovial fluid could reflect cartilage alteration in arthritis in the horse.     

Regeneration of cartilage tissue by autologous chondrocytes transplantation for cartilage defects in a experimental bovine model

To evaluate the effects of chondrocytes transplantation on the regeneration of cartilage by intraarticular injection or injection into blood clots at cartilage defects, eight full-thickness cartilage defects were created surgically on the articular surface of each femoral trochlea of two calves. Autologous chondrocytes were isolated individually from the cartilage pieces collected at the creation of defects. And isolated cells were cultured in monolayers for proliferation. Cells were injected into synovial fluid (Group 2, n=11) or into the blood clots at the cartilage defects (Group 3, n=5) of the left femoropatellar joint on weeks 2 and 3, respectively after the operation. The defects (Group 1, n=16) of right femoropatellar joint were left untreated in the control group. After 14 weeks, repaired tissues were evaluated based on gross and histological examinations. In Group 3, more repaired tissues and a better interface between the repaired tissue and host cartilage were observed compared with the results for Groups 1 and 2. Moreover, cartilaginous tissue were observed more in defects of Group 3 than in defects of other groups. In conclusion, the present study suggests that the injection of cells into the blood clot at a cartilage defect might be applicable for the regeneration of damaged cartilage.     

Meniscal Release in Cruciate Ligament Intact Stifles Causes Lameness and Medial Compartment Cartilage Pathology in Dogs 12 Weeks Postoperatively

Objective— To evaluate after 12 weeks the effects of caudal medial meniscal release (MR) in the cranial cruciate ligament-intact canine stifle.
Study Design— Blinded, prospective in vivo study.
Animals— Purpose-bred hound dogs (n=10).
Methods— Either MR (n=5) or a sham (SH) surgery (n=5) was performed via arthroscopy. Orthopedic examination and subjective lameness evaluation were performed in each dog preoperatively and at 4, 8, and 12 weeks after surgery. Twelve weeks postoperatively, ultrasonographic, radiographic, and arthroscopic examinations were performed on the operated stifles. Gross pathology of the articular cartilage, cruciate ligaments, and menisci was assessed. India ink staining of the femoral and tibial articular surfaces was performed to determine the percent area of articular cartilage damage.
Results— At 8 and 12 weeks after surgery, MR dogs were lamer than SH dogs. At 12 weeks, the degree of radiographic OA was significantly higher in MR stifles than in SH stifles. Gross and sonographic meniscal pathology was more severe in MR stifles compared with SH stifles. MR stifles had significantly more severe articular cartilage pathology compared with SH stifles 12 weeks after surgery; pathology was most severe in the medial compartment.
Conclusions— MR alone is associated with articular cartilage loss, further meniscal pathology, degenerative joint disease, and lameness.
Clinical Relevance— Subsequent osteoarthritis and dysfunction of the stifle joint should be considered when making clinical decisions regarding MR in dogs.     

Chondrocyte apoptosis and decrease of glycosaminoglycan in cranial cruciate ligament insertion after resection in rabbits

We examined time-dependent histological changes of the calcified fibrocartilage area in a tibial cranial cruciate ligament (CCL) insertion after ligament resection in rabbits. The animals were divided into two groups: those undergoing CCL substance resection in the right stifle (resected group) and those receiving the same operation without CCL resection in the left stifle (sham operated group). Five animals were euthanized with deep anaesthesia at four time periods (1, 2, 4 and 6 weeks), and Haematoxylin-eosin and Safranin-O stainings and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) staining were performed. The average percentage of TUNEL-positive chondrocytes and the average thickness of the glycosaminoglycan (GAG)-stained area in the calcified fibrocartilage area were measured. Two and 4 weeks after the surgery, the average percentages of TUNEL-positive chondrocytes in the resected group (23.8 +/- 10.3% and 15.9 +/- 6.7%, respectively) were significantly higher than those in the sham operated group (8.9 +/- 3.8% and 7.4 +/- 1.6%, P<0.05, respectively). Six weeks after the surgery, the average thickness of the GAG-stained area in the resected group (7.7 +/- 13. 5 microm) was significantly smaller than that in the sham operated group (69.4 +/- 39.9 microm, P<0.05). Our results suggest that the average percentage of TUNEL-positive chondrocytes became a peak in 2 weeks and that histological changes occurred in 6 weeks. The chondrocyte apoptosis can induce decrease of GAG-stained area after resection of CCL. Therefore, chondrocyte apoptosis in the calcified cartilage area in the CCL tibial insertion might lead to histological changes.     

Evaluation of a Collagenase Generated Osteoarthritis Biomarker in the Synovial Fluid from Elbow Joints of Dogs with Medial Coronoid Disease and Unaffected Dogs

Objective— To evaluate whether synovial fluid concentrations of an osteoarthritis biomarker in dysplastic canine elbows with medial coronoid disease (MCD) are elevated compared with unaffected elbows and to determine if these concentrations correlate to the degree of articular cartilage damage.
Study Design— Cross sectional clinical study.
Animals— Dogs (n=19; 35 elbows) with MCD and dogs (8; 16 elbows) with unaffected elbows.
Methods— Concentrations of a collagenase-generated cleavage neoepitope of type II collagen (Col2-3/4C_{long mono}, or C2C) in joint fluid from elbows were analyzed and compared between dogs with MCD and unaffected dogs. Correlation of C2C concentration with subjective grading of articular cartilage surface damage was also evaluated.
Results— Mean (±SD) C2C concentration from MCD dogs was significantly higher (112.3±24.8 ng/mL) than in unaffected dogs (76.1±16.9 ng/mL; P <.05). There was a moderate correlation between cartilage damage grade and increasing C2C concentrations ( P <.05, r=0.62)
Conclusion— C2C concentrations are elevated in the synovial fluid of dogs with MCD compared with unaffected elbows, and a moderate, significant correlation was identified between these concentrations and subjective grading of articular cartilage damage.
Clinical Relevance— This preliminary data suggest that C2C concentrations in synovial fluid may have potential as a biomarker for diagnosis of articular cartilage damage associated with MCD and as a means of objectively determining the degree of articular cartilage damage.     

Evaluation of avocado and soybean unsaponifiable extracts for treatment of horses with experimentally induced osteoarthritis

OBJECTIVE: To evaluate the use of a combination of avocado and soybean unsaponifiable (ASU) extracts for the treatment of experimentally induced osteoarthritis in horses. ANIMALS: 16 horses. PROCEDURES: Osteoarthritis was induced via osteochondral fragmentation in 1 middle carpal joint of each horse; the other joint underwent a sham operation. Horses were randomly allocated to receive oral treatment with ASU extracts (1:2 [avocado-to-soybean] ratio mixed in 6 mL of molasses; n = 8) or molasses (6 mL) alone (placebo treatment; 8) once daily from days 0 to 70. Lameness, response to joint flexion, synovial effusion, gross and histologic joint assessments, and serum and synovial fluid biochemical data were compared between treatment groups to identify effects of treatment. RESULTS: Osteochondral fragmentation induced significant increases in various variables indicative of joint pain and disease. Treatment with ASU extracts did not have an effect on signs of pain or lameness; however, there was a significant reduction in severity of articular cartilage erosion and synovial hemorrhage (assessed grossly) and significant increase in articular cartilage glycosaminoglycan synthesis, compared with placebo-treated horses. CONCLUSIONS AND CLINICAL RELEVANCE: Although treatment with ASU extracts did not decrease clinical signs of pain in horses with experimentally induced osteoarthritis, there did appear to be a disease-modifying effect of treatment, compared with findings in placebo-treated horses. These objective data support the use of ASU extracts as a disease-modifying treatment for management of osteoarthritis in horses.     

EFFECT OF HYDRATION ON SIGNAL INTENSITY OF GELATIN PHANTOMS USING LOW-FIELD MAGNETIC RESONANCE IMAGING: POSSIBLE APPLICATION IN OSTEOARTHRITIS

Five gelatin phantoms were constructed to study the effect of matrical hydration on magnetic resonance imaging (MRI) signal intensity using a low-field strength imager. Water content of the phantoms ranged from 75 to 95% weight/weight. Signal intensity values of each phantom were measured using five imaging sequences: proton density, T1-weighted, T2-weighted, inversion recovery with short inversion time, and inversion recovery with long inversion time. There was significant positive correlation (p < .05) of signal intensity with differences in hydration using the T2-weighted sequence and the inversion recovery sequence with short inversion time. Significant negative correlations (p < .05) were found with T1-weighted imaging and the inversion recovery sequence with long inversion time. In a second part of the study, in vivo focal variations in MRI signal intensity were evaluated in a canine cranial cruciate ligament deficient model of osteoarthritis. Signal intensity measurements were obtained from multiple areas of articular cartilage to identify an initial stage in osteoarthritis that is characterized in part by increased hydration of articular cartilage. At 6 weeks post-transection of the cranial cruciate ligament, an increase in signal intensity was detected in the articular cartilage of the weight-bearing portion of the lateral femoral condyle and the caudal portion of the medial tibial condyle with T1-weighted imaging. The increase in signal intensity may reflect increased proteoglycan synthesis by chondrocytes that also occurs early in the pathogenesis of osteoarthritis.