The Effect of Exercise on the Healing of Articular Cartilage Defects in the Equine Carpus

Arthroscopic surgery was performed on 12 horses (2-4 years of age) to create a 7 x 14 mm full-thickness cartilage defect in one radial carpal bone and in the contralateral third carpal bone. Six horses remained confined to a small paddock and six horses underwent a program of increasing exercise consisting of walking, trotting, and cantering for 13 weeks. All lesions showed evidence of healing at week 6 that progressed to more complete healing at week 13. There was no difference in the amount of repair tissue covering the defect. Histologically, the lesions healed with a combination of fibrous tissue and fibrocartilage. The repair tissue was significantly thicker in the exercised horses but there was no difference in repair quality. It was concluded that radial carpal and third carpal lesions have an equal ability to heal and that early postoperative exercise is not detrimental to the repair tissue within these carpal cartilage defects.     

Study on Inducing Equine Bone Marrow Mesenchymal Stem Cells Differentiated into Chondrocytes

This study was aimed to establish equine bone marrow mesenchymal stem cells(BMSCs) line and induce it to differentiate into chondrocytes. The BMSCs were collected by cutting the bone and flushing the cutting surface by PBS, and then the bone marrow was washed with PBS,the collected cells were cultured after centrifugation. The cells were purified by passaging,the stem cell properties were tested before the induction. And the cells were also appraised by the expression of the special gene of chondrocytes as well as staining the differentiated cells with Alcian blue to insure the induction was effective. The obtained BMSCs expressed Sox2 and Nanog genes, which were the stem cell special genes, and also expressed CD44, CD90 and CD105 genes, but absent of CD34 and CD45 genes. The shapes of the 3rd passage BMSCs were changed after cultured in inducing medium for a few days. Furthermore, the cells were positive to Alcian blue staining, increased expression of the Col special gene of chondrocyte day by day as well. According to this study, the BMSC line was established, and the BMSCs were induced and differentiate into chondrocytes successfully.     

马骨髓间充质干细胞向软骨细胞的诱导分化

试验旨在建立马骨髓间充质干细胞（bone marrow mesenchymal stem cells,BMSCs）细胞系,并诱导其向软骨细胞分化。通过获取马BMSCs,进行细胞培养和纯化,对第3代（P3）细胞进行干细胞特性鉴定,并诱导其向软骨细胞分化,对分化后的细胞染色,并检测其软骨细胞特异性基因的表达。结果显示,获得的马BMSCs表达标记基因Sox2和Nanog,并表达间充质干细胞表面标记因子CD44、CD90和CD105,不表达造血细胞表面标志物CD34和CD45。P3代细胞经诱导培养后形态发生改变,阿尔新蓝染色为阳性,并表达软骨细胞特异基因Col,且其表达量随着诱导分化时间的增加而增高。综上表明,本试验建立了马BMSC细胞系,并成功诱导其分化为软骨细胞,为软骨损伤的干细胞治疗提供了试验依据。     

The Effect of Glucosamine and Chondroitin on Stressed Equine Cartilage Explants

Effective prevention and treatment of osteoarthritis for horses is still needed. This research tests the ability of glucosamine and chondroitin sulfate (GLN/CS) to mitigate inflammatory and mechanical stress in vitro. In this study, GLN/CS mediate this effect by a decrease of the synthesis of nitric oxide (NO) and a decrease of proteoglycan release from the extracellular matrix in stressed cartilage explants. Explants were cultured with interleukin-1 (IL-1) + mechanical trauma with and without GLN/CS. NO and prostaglandin E2 were measured as indicators of an inflammatory response. Glycosaminoglycans were measured as an indicator of cartilage breakdown. NO levels in the stressed explants with GLN/CS treatment were lower than the IL-1 + mechanical impact treatment alone and did not differ from control group. The glycosaminoglycan release was also lower in the GLN/CS treatment than the IL-1 + mechanical impact treatment, although the prostaglandin E2 concentration was not affected. This study offers some evidence that GLN/CS treatment can partially mitigate the catabolic response to inflammatory stress and mechanical trauma in equine cartilage explants. These results provide additional support for the continued study on the benefit of GLN/CS for horses with cartilage degeneration.     

Treatment of Bilateral Medial Femoral Condyle Articular Cartilage Fissures in a Horse Using Bone Marrow-Derived Multipotent Mesenchymal Stromal Cells

The objective of this study was to describe the use, and outcome, of multipotent mesenchymal stromal cells (MSCs) in the treatment of equine articular cartilage defects of the medial femoral condyle. A 4-year-old Thoroughbred gelding (n = 1) with bilateral stifle athroscopy was found to have bilateral articular cartilage fissure defects of the medial femoral condyles with concurrent cranial cruciate ligament injury. Bone marrow derived MSCs were isolated, expanded, and suspended in a partially autologous fibrin glue. The initial cell/fibrin glue mixture was delivered arthroscopically into the articular cartilage defects 90 days after the initial arthroscopic examination. Follow-up treatments included two additional injections of MSCs suspended in lactated Ringers solution, 5 and 13 months after the initial examination, directly into the joint. Post-treatment outcome was assessed by arthroscopic examination and by comparison of preinjury and post-treatment performance records. Arthroscopic evaluation 4 months after the initial MSC treatment revealed marked smoothing, reduction in the depth of cartilage defects and observation of moderate improvement in the cranial cruciate ligament. Approximately 15 months after the initial MSC treatment the horse returned to racing. Analysis of race records demonstrated that the post-treatment (including all three MSC treatments) average race earnings (earnings per start) were comparable with those predating the initial injury. The favorable clinical response in the face of an unknown, but likely, guarded prognosis suggest that MSC therapy is not deleterious and may augment healing of articular cartilage fissures of the medial femoral condyle. MSCs represent a viable and promising alternative therapy in the treatment of articular cartilage injuries in performance horses.     

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。     

The Effect of Trans-Stifle External Skeletal Fixation and Hyaluronic Acid Therapy on Articular Cartilage in the Dog

Transarticular external skeletal (TES) fixators were applied unilaterally to the stifle joints of 10 young adult dogs. After 4 weeks, the fixators were removed from all dogs. Two dogs were not allowed a remobilization period, whereas 8 dogs were provided with 4 additional weeks of weight-bearing activity in a kennel run. Four dogs were given high-molecular weight hyaluronic acid by intra-articular injection weekly during the remobilization period. Clinical gait evaluations and range of motion were determined during the remobilization period. Articular cartilage samples from both stifle joints of all dogs were evaluated histologically and histochemically. No significant differences in gait scores or range of motion were noted between treated and untreated dogs. Articular cartilage proteoglycan content was reduced after 4 weeks of trans-stifle external skeletal fixation as determined by loss of alcian blue (AB) histochemical staining. Improved homogeneity of histochemical staining was observed after remobilization. However, remobilization was associated with histological damage to the surface and tangential layers of articular cartilage. Remobilization combined with hyaluronic acid (HA) therapy improved histochemical staining and reduced structural damage to articular cartilage when compared with remobilization alone.     

用快中子辐照的碳纤维编织物诱发关节软骨再生

采用快中子辐照的碳纤维编织物，填充到实验兔膝关节软骨面缺损处，将实验兔分期分批剖杀，用肉眼、光学显微镜和扫描电子显微镜对再生的部分进行了观察。结果表明，碳纤维编织物可以诱发软骨组织再生，以其作为填充物是治疗因软骨损伤而导致的关节僵直的理想生物材料。     

Effect of Dimethylsulfoxide on Articular Cartilage Proteoglycan Synthesis and Degradation, Chondrocyte Viability, and Matrix Water Content

Objective—To determine the effects of dimethylsulfoxide (DMSO) exposure on cartilage proteoglycan (PG) synthesis, PG degradation, chondrocyte viability, and matrix water content. Study Design—Using a cartilage explant culture system, PG synthesis, PG degradation, matrix water content, and chondrocyte viablity were determined for cartilage exposed to DMSO daily for selected periods of time. Animals or Sample Population—Juvenile bovine (calf) carpometacarpal joint cartilage ex-plants. Methods—PG synthesis: Explants (n = 30/group) were separated into 10 groups based on the time of daily exposure to 10% DMSO. Exposure time was repeated daily for 3 days. The control group was incubated in basal medium alone for 3 days, with daily medium changes. Once all DMSO exposure times were complete for the third day, PG synthesis was determined by analysis of incorporation of radiolabelled sulfate. Cell viability: Explants (n = 3/group) were subjected to an identical DMSO exposure protocol, and examined histologically. The percentage of viable cells/high power field (hpf) was calculated for each group. PG degradation: Explants (n = 21/group) were preincubated with radiolabelled sulfate, then subjected to a similar DMSO exposure protocol. The medium was collected from all explants daily and assayed for PG content. After 3 days, the explants were digested and total labelled PG content determined. Percent of total explant labelled PG content released into the medium daily was determined for each group. Water content: Explants (n = 21/group) were separated into three treatment groups, one of which had no treatments performed, whereas the other two groups were incubated in basal medium for 72 hours, one with, and one without, 10% DMSO. Wet and dry weights were determined, and percent water calculated, for all three groups. Separate 1-way ANOVA were performed, with appropriate post hoc tests (p < .05). Results—PG synthesis was significantly lower than control for all time periods of DMSO exposure except for 1 and 3 hours, and decreased in a time-dependent manner after the 1-hour exposure time. The mean percentage of viable cells/hpf was significantly lower than control for the 1-, 3-, 9-, 12-, and 24-hour treatment groups. There was no significant difference in PG degradation for any group compared with control for the first 2 days of incubation. All groups except the 24-hour group had a significantly higher degradation compared with control for the third day of incubation. Cartilage exposed to DMSO for 72 hours had a significantly lower water content, and cartilage incubated in basal medium alone for 72 hours had a significantly higher water content than cartilage that received no DMSO and no incubation. Conclusions—DMSO, in relatively low concentration, is detrimental to articular cartilage PG     

Clinical Experience Using MicroLactin for the Treatment of Equine Inflammatory Disease

MicroLactin is a patented milk protein concentrate whose mode of action is proposed to inhibit neutrophil activation in inflammation and to bolster the immune response in musculoskeletal diseases. MicroLactin was empirically used in the treatment of a series of equine clinical cases. MicroLactin was given in two trials to 166 horses in which neutrophils were associated with an inflammatory response. The primary clinical groups having the greatest positive responders to the use of MicroLactin were: respiratory (92%), joint lameness/foot trauma (90%), muscle injury/myositis (92%), equine protozoal myeloencephalitis (EPM) (81%), skin trauma/hypersensitivity (89%), and toxic enteritis (89%). Positive clinical results were seen within 10 to 14 days when MicroLactin was used as a daily treatment either alone or in combination with other anti-inflammatory agents or as an adjunct to the primary treatment.     

Arthroscopic Surgery for the Treatment of Osteochondrosis in the Equine Shoulder Joint

Osteochondritis dissecans (OCD) and subchondral cyst-like lesions in 13 shoulders of 11 horses were treated arthroscopically by curettage and lavage. Lameness decreased in all 11 horses. Nine horses were sound, five of them athletically sound, after 5 to 20 months. Complications included the development of subchondral cyst-like lesions and signs of degenerative joint disease. Arthroscopic surgery of the equine shoulder can be done through two portals, one for the arthroscope and one for an instrument. A few hand instruments such as a probe, Ferris-Smith rongeurs, and small, large, and right-angled curettes are needed to debride most lesions. Motorized equipment can expedite the process.     

Arthroscopic Surgery for the Treatment of Osteochondritis Dissecans in the Equine Femoropatellar Joint

Forty limbs with femoropatellar osteochondritis dissecans in 24 horses were treated with arthroscopic surgery. Lesions were bilateral in 16 horses and unilateral in eight horses. Diagnostic examination and surgical treatment were performed through a single arthroscopic portal; five different instrument portal locations and six instrument approaches were used. Lesions were localized to the lateral trochlear ridge of the femur in 31 affected joints, medial trochlear ridge in two joints, lateral and medial trochlear ridges together in two joints, lateral trochlear ridge plus patella in four joints, and patella alone in one joint. The lesions consisted of subchondral defects containing chondral or osteochondral flaps or fragments, or were seen as dimpling, cracking, fibrillation, or erosion of articular cartilage, or intact cartilage over a subchondral defect. Loose bodies were found in three joints. There was a poor correlation between radiologic and arthroscopic findings. Surgical manipulations included removal of flaps, fragments, and undermined articular cartilage, and debridement of the subchondral defect. Three horses were euthanized: one electively to assess the joint grossly, one because of complications following surgery and salmonellosis, and one because of unrelated forelimb abnormalities. Immediate clinical improvement after surgery was seen in the 22 horses permitted to survive. Long-term follow-up on seven of 10 racehorses revealed that two have raced successfully, two are "ready to race," three are training sound, two are sound at pasture (still in convalescence), and one has been reoperated. Of six horses used for show or pleasure, three are being shown sound, one is sound for pleasure, and two are training sound. The remaining horses are convalescing.     

Arthroscopic Anatomy of the Equine Femoropatellar Joint and Approaches for Treatment of Osteochondritis Dissecans

Arthroscopic approaches to the femoropatellar joint were developed to determine their usefulness for evaluation and surgical treatment of osteochondritis dissecans. It was found that the articular cartilage of the lateral trochlear ridge, medial trochlear ridge, intertrochlear groove, patella, and the lateral and medial reflections of the joint capsule could be examined from an infrapatellar arthroscopic portal. The suprapatellar pouch could be examined partially. Lateral and medial instrument portals were evaluated to determine the accessibility of the lateral and medial trochlear ridges of the femur in the areas where osteochondritis dissecans lesions frequently occur. Sliding the arthroscope sleeve beneath the patella when entering the joint was associated with iatrogenic cartilage lesions. A new technique that directed the arthroscope lateral to the lateral trochlear ridge eliminated iatrogenic cartilage damage.     

马病毒性动脉炎诊断技术研究进展

马病毒性动脉炎(EVA)是马属动物之间通过呼吸道或生殖器官传播的传染病,是危害养马业及赛马比赛的重要疾病之一,应用各种诊断技术对该病做出准确的诊断是预防和控制该病的前提.EVA的诊断技术包括检测病毒,检测血清抗体,涉及的技术和方法包括病毒的分离及中和试验、血凝及血凝抑制试验、免疫荧光试验以及酶联免疫吸附试验等.每种诊断技术都有其各自的优势和局限性,应根据目的及工作条件等综合情况予以选择,论文就各种诊断技术的原理、方法及特点进行了概述和比较,并对今后的应用前景和发展趋势进行了展望.