猪喉软骨硫酸软骨素的制备和自由基清除活性

直接用木瓜蛋白酶水解软骨,三氯醋酸除蛋白质,乙醇沉淀干燥得硫酸软骨素粗多糖,经DEAE-Sepharose fast flow离子交换层析分离纯化,高效凝胶渗透色谱和琼脂糖凝胶电泳测定其相对分子质量和纯度,比较硫酸软骨素粗品和纯品的体外自由基清除活性。结果显示硫酸软骨素粗品和纯品提取率分别为31.56±0.46%和19.79±0.78%,后者相对相对分子质量为75 174,粗品的DPPH.和.OH清除活性最强,随着产品纯度的提高,活性降低,而对于O2-.清除活性结果则相反。     

Safety of, and biological and functional response to, a novel metallic implant for the management of focal full-thickness cartilage defects: Preliminary assessment in an animal model out to 1 year.

Focal full-thickness cartilage lesions of the human medial femoral condyle (MFC) can cause pain and functional impairment. Affected middle-aged patients respond unpredictably to existing treatments and knee arthroplasty may be required, prompting risk of revision. This study assesses the safety of, and biological and functional response to, a metallic resurfacing implant which may delay or obviate the need for traditional arthroplasty. The anatomic contour of the surgically exposed MFC of six adult goats was digitally mapped and an 11 mm diameter full-thickness osteochondral defect was created. An anchor-based Co-Cr resurfacing implant, matching the mapped articular contour, was implanted. Each goat's contralateral unoperated femorotibial joint was used as a control. Postoperative outcome was assessed by lameness examination, radiography, arthroscopy, synoviocentesis, necropsy, and histology up to 26 (n = 3) or 52 (n = 3) weeks. By postoperative week (POW) 4, goats demonstrated normal range of motion, no joint effusion, and only mild lameness in the operated limb. By POW 26 the animals were sound with only occasional very mild lameness. Arthroscopy at POW 14 revealed moderate synovial inflammation and a chondral membrane extending centrally across the implant surface. Radiographs at POWs 14 to 52 implied implant stability in the operated joints, as well as subchondral bone remodeling and mild exostosis formation in the operated and contralateral unoperated joints of some goats. By POW 26, histology revealed new trabecular bone abutting the implant. At POWs 26 and 52 MFC cartilage was metachromatic and intact in the operated and unoperated femorotibial joints. Proximal tibiae of some operated and unoperated limbs demonstrated limited subchondral bone remodeling and foci of articular cartilage fibrillation and thinning. The chondral membrane crossing the prosthesis possessed a metachromatic matrix containing singular and clustered chondrocytes. Our data imply the safety, biocompatibility, and functionality of the implant. Focal articular damage was documented in the operated joints at POWs 26 and 52, but lesions were much reduced over those previously reported in untreated defects. Expanded animal or preclinical human studies are justified.     

兔关节软骨细胞的分离、培养和形态学特征

[目的]探讨兔关节软骨细胞的分离、培养方法,观察单层高浓度培养时细胞表型表达情况.[方法]无菌条件下,从 2周龄新西兰白兔的颞颌关节及四肢关节髁突面削取软骨片,采用机械-酶消化法分离软骨细胞,经台盼蓝拒染计数,将细胞按 1× 106个 /孔接种于 6孔培养板,传代培养,描绘生长曲线.利用相差显微镜及透射电镜观察细胞形态.应用甲苯胺蓝及Ⅱ型胶原免疫组化对细胞进行鉴定.[结果]每克软骨能获取 1 5× 106个软骨细胞,活性率为 95%.培养 2～ 3 d,细胞贴壁、变形,呈多角形; 8 d左右,细胞融合成层.透射电镜观察显示细胞核圆形,有丰富的粗面内质网、高尔基体及分泌的基质成分.甲苯胺蓝及Ⅱ型胶原染色阳性.细胞传至 5代后,出现"成纤维细胞样".[结论]本研究建立了简单易行的软骨细胞分离、培养方法;初代、第 2代细胞生长良好,适合于实验研究;软骨细胞 5代培养后,细胞表型发生改变.     

Synovial chondromatosis of four compartments of the knee: medial and lateral tibiofemoral spaces,patellofemoral joint and proximal tibiofibular joint

A case of synovial chondromatosis of the proximal tibiofibular joint in addition to lateral and medial tibiofemoral spaces and patellofemoral joint has been presented.     

软骨下骨钻孔术对兔软骨缺损修复的远期效果观察

目的　观察软骨缺损的修复过程 ,比较不同数目钻孔术对软骨缺损修复的远期效果。方法　用中国白兔 4 0只 ,在股骨髁关节面造成 6mm× 8mm全层软骨缺损 ,分别施行 10孔及 5孔钻孔术 ,孔径1mm ,于术后 12周及 13个月取材 ,做组织学及电镜观察 ,并进行评估。结果　 (1) 12周时 10孔、5孔和对照组的修复组织中类透明软骨分别占 70 %、5 0 %、0 % ;13个月时 10孔、5孔和对照组的修复组织中类透明软骨分别占 90 %、10 0 %、0 %。 (2 )修复组织厚度 :12周时 10孔及 5孔组均明显高出毗邻软骨 ,且 10孔明显高于 5孔 ;13个月时 10孔与 5孔无显著性差异 ,且已接近毗邻软骨厚度。 (3)修复组织覆盖缺损的面积 :12周时 10孔 >5孔 >对照组 ;13个月时 10孔与 5孔无显著性差异 ,但二者均明显大于对照组。结论　软骨下骨钻孔对关节软骨缺损修复的远期效果良好 ,能长期适应关节的生理运动和功能负重 ,10孔与 5孔的远期修复效果无显著性差异。     

特异性COX-2抑制剂保护骨关节炎软骨的临床观察

目的探讨特异性COX-2抑制剂对骨关节炎的关节软骨保护作用的临床研究。方法门诊选取膝关节骨关节炎患者80例,随机分为特异性COX-2抑制剂组和维骨力对照组,各40例,单盲给药。应用平均Womac关节炎指数评分和平均OA严重程度指数评定两组用药前后临床症状改善情况及使用血清软骨代谢标志物蛋白聚糖与型胶原和关节MRI进行检测关节软骨结构与功能的改变情况,并作统计学分析。结果经12个月治疗观察,特异性COX-2抑制剂组的平均Womac关节炎指数评分和平均OA严重程度指数在前3个月改善明显优于维骨力对照组(P<0.01),但在治疗12个月后,两组差异不显著(P>0.05);治疗12个月后特异性COX-2抑制剂组的软骨代谢指标蛋白聚糖和型胶原及关节MRI均显示组内变化显著(P<0.01),而与维骨力组对比结果无显著性差异(P>0.05)。结论特异性COX-2抑制剂对骨关节炎的软骨有保护和促进修复作用,兼有抗炎与保护软骨双重疗效,它是一类既能改善关节炎症状又能改善关节软骨结构的药物,值得临床广泛应用。同时本研究为其在临床应用和进一步研究此类药物提供了更好的理论基础和依据。     

High-resolution imaging of progressive articular cartilage degeneration.

The objective of this study was to develop and verify a new technique for monitoring the progression of osteoarthritis (OA) by combining a rat model with the imaging modality optical coherence tomography (OCT). Time-sequential, in vivo, OCT imaging was performed on the left femoral condyles of 12 Wistar rats following sodium-iodoacetic acid-induced OA progression. The right femoral condyles (untreated) were also imaged and served as controls. Imaging was performed on days 0, 10, 20, 30, and 60 with an OCT system capable of acquiring images at four frames per second and an axial resolution of 5 microm. Progressive changes were analyzed using an OA scoring system. OCT successfully identified progressive cartilage degeneration as well as alteration of the cartilage/bone interface. Significant changes to both of these structures were observed in the sodium-iodoacetic acid-injected condyles. Structural changes detected with OCT were confirmed histologically. OCT in combination with a well-known model used in arthritis research represents a powerful tool for following degenerative joint disease progression in a given animal by detecting changes to the cartilage/bone interface and articular cartilage.     

鸡胚股骨无血清培养研究软骨形成和骨形成

为探讨软骨形成和骨形成的作用机制，用孵化14天鸡胚股骨进行无血清培养，对其细胞增殖分化和基质矿化情况进行观察。结果显示：股骨在培养期间长度增长，骺端增大。软骨细胞、成骨细胞增殖分化成熟;成骨细胞分泌基质小泡和类骨质，并有奶盐沉积。提示鸡胚股骨在无血清培养条件下，可在体外进行软骨形成和骨形成，此模型可用来进行骨形成研究。