骨性关节炎软骨下骨病变机制研究进展

长期以来对骨性关节炎(osteoarthritis,OA)中关节软骨的改变进行较为深入的研究,但对软骨下骨的改变研究相对较少,而软骨下骨的改变是OA进程中所必需的病理变化.近年研究发现软骨下骨改变在OA发病过程中起着积极作用,软骨下骨硬化或软化不仅是OA发生的结果,而且与其发生发展密切相关,提示OA治疗既应关注软骨变化,又要预防软骨下骨退变.该文从软骨下骨的生物力学、骨重塑、分子生物学等方面就软骨下骨在骨性关节炎发病中的病变机制研究进展作一综述.     

膝骨性关节炎患者胫骨软骨下骨超微结构改变的CT评价

文题释义： 膝骨性关节炎：是一种以渐进性的关节损伤为主要特征,并最终导致患者关节疼痛甚至残疾,极大地降低患者生活质量。 CT：利用精确准直的X射线束、γ射线、超声波等,与灵敏度极高的探测器一同围绕人体的某一部位作一个接一个的断面扫描,具有扫描时间快、图像清晰等特点。 背景：软骨下骨的改变在膝骨性关节炎的作用得到了越来越多的重视,但是既往研究主要以动物为观察对象,由于动物与人存在差异,因而直接在人体关节内获得相关数据是非常必要的。 目的：通过CT技术评价非膝骨性关节炎患者与膝骨性关节炎患者软骨下骨板及软骨下骨超微结构的区别,来探讨软骨下骨在膝骨性关节炎疾病的发生和发展中的作用。 方法：于2016年7月至2018年7月在郑州市骨科医院影像科就诊患者中,收集30例膝骨性关节炎患者(膝骨性关节炎组)及30例非膝骨性关节炎患者(非膝骨性关节炎组)的CT扫描数据,使用MIMICS软件比较2组胫骨平台内外侧软骨下骨板以及软骨下骨小梁超微结构。试验于2016-06-10经郑州市骨科医院伦理委员会审批通过,审批号为2016医院伦审第004号。 结果与结论：①与非膝骨性关节炎组相比,膝骨性关节炎组软骨下骨板在外侧部位和内侧部位骨密度都明显增加,孔隙率则出现显著的下降,而软骨下骨板厚度内侧部分较非膝骨性关节炎组显著增厚;②膝骨性关节炎软骨下骨小梁同样存在明显变化,表现为膝骨性关节炎组内外侧软骨下骨骨小梁厚度较非膝骨性关节炎组均明显增加,同时内侧松质骨分离度也较非膝骨性关节炎组低;膝骨性关节炎组结构模型指数和连接密度值低于非膝骨性关节炎组;③结果表明,膝骨性关节炎患者胫骨软骨下骨板及软骨下骨松质骨的改变主要在于超微结构稳态的破坏,这一改变可能是膝骨性关节炎发病原因之一。 ORCID: 0000-0002-9805-3084(白玉) 中国组织工程研究杂志出版内容重点：人工关节;骨植入物;脊柱;骨折;内固定;数字化骨科;组织工程     

软骨下骨病理骨重建过程中骨细胞的代谢机制

背景：骨性关节炎一直被认为是关节软骨的退变和降解。现已逐步认识到软骨下骨结构及分子代谢的病理变化在关节退变的过程中发挥了重要的作用。 目的：观察软骨下骨在骨性关节炎进程中结构及细胞、相关因子和信号通道的变化,分析软骨下骨的病理改变对关节软骨退变的影响。 方法：使用骨性关节炎、软骨下骨、软骨退变、骨重建、骨代谢等关键词,检索中国知识资源总库与PubMed、Medline、Embase数据库中的相关研究论文,了解骨性关节炎进程中软骨下骨骨代谢平衡破坏后在结构及分子信号机制的病理变化,分析这些病理改变与关节软骨退变的关系。 结果与结论：纳入 54篇符合标准的文献。软骨下骨的病理骨重建贯穿骨性关节炎的始终,局部骨细胞的代谢及干细胞分化异常与软骨退变相互影响。通过对软骨下骨病理骨重建过程中骨细胞代谢机制的深入了解,为骨性关节炎提供了新的治疗靶点。     

淫羊藿苷治疗骨性关节炎过程中对关节软骨、软骨下骨、滑膜等影响的机制

文题释义： 淫羊藿苷：为淫羊藿干燥茎叶的提取物,呈淡黄色针状结晶粉末, 相对分子质量为676.65, 属黄铜类化合物,现代药理学研究发现其具有很强的生物活性, 对骨组织、免疫系统、肿瘤组织、神经系统、生殖系统、内分泌系统和心血管系统等具有显著作用。 滑膜：是关节囊的内层。滑膜呈淡红色,平滑闪光,薄而柔润,由疏松结缔组织组成,其功能是制造和调节滑液等。滑膜直接附着于关节软骨的边缘并向内贴附在关节囊内的非关节区域,覆盖在关节囊、关节内韧带、骨与肌腱表面。滑膜分泌滑液,在关节活动中起重要作用。背景：淫羊藿苷是具有补肾强筋健骨功效的淫羊藿的主要有效成分,近年来大量的研究发现淫羊藿苷在治疗骨性关节炎方面有着显著作用。 目的：综述淫羊藿苷治疗骨性关节炎的分子机制研究进展。 方法：第一作者应用计算机以“Icariin、Osteoarthritis、Cartilage、Subchondral bone、Synovial membrane 、Synovium、Inflammation”及“淫羊藿苷、骨关节炎、骨性关节炎、软骨、软骨下骨、滑膜、炎症”作为主题词检索PubMed、中国知网、万方、维普等数据库相关文献,按入选标准及排除标准进行筛选,最终纳入42篇文献进行分析。 结果与结论：淫羊藿苷通过促进骨髓间充质干细胞的成软骨分化及增强软骨细胞和成骨细胞的增殖,抑制软骨细胞外基质的降解、降低破骨细胞的活性和减轻炎症因子所致的滑膜炎症反应来有效的治疗骨性关节炎。但其最佳有效剂量及浓度安全性仍需要大量实验研究,目前绝大部分实验仍停留在动物及组织细胞等基础实验,尚需要大量临床研究,继续完善其具体机制,以期为淫羊藿苷治疗骨关节炎提供循证医学证据。ORCID:0000-0002-2013-743X(余绍涌)中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

透骨消痛胶囊干预膝骨性关节炎软骨下骨重塑的分子机制**☆

背景：前期研究表明透骨消痛胶囊能有效治疗膝骨性关节炎,且对膝关节软骨有保护作用,软骨下骨重塑在骨性关节炎病理进程中起重要作用。 目的：观察透骨消痛胶囊干预骨性关节炎软骨下骨重塑的作用及并探讨其作用机制。 方法：新西兰大白兔分为正常组、模型组、壮骨关节丸组、透骨消痛胶囊组4组。除正常组外动物均按改良Hulth法造模。正常组、模型组兔每天给予生理盐水灌胃;壮骨关节丸组兔每天给予壮骨关节丸水溶液灌胃;透骨消痛胶囊组兔每天给予透骨消痛胶囊水溶液灌胃。 结果与结论：造模后6,9,12周,透骨消痛胶囊组软骨下骨Cyclin D1基因、胰岛素样生长因子1、细胞核因子κB受体活化因子配体mRNA表达均显著高于正常组(P < 0.05)。与模型组比较,造模后1周,透骨消痛胶囊组Cyclin D1 mRNA表达较低(P < 0.05);造模后6周,透骨消痛胶囊组细胞核因子κB受体活化因子配体表达较低(P < 0.05);造模后9,12周,透骨消痛胶囊组各项指标表达均较低(P < 0.05)。与壮骨关节丸组比较,造模1周后,透骨消痛胶囊组Cyclin D1 mRNA表达较低(P < 0.05);6周时透骨消痛胶囊组细胞核因子κB受体活化因子配体表达较低(P < 0.05);9,12周后,透骨消痛胶囊组胰岛素样生长因子1 mRNA表达较低(P < 0.05)。提示透骨消痛胶囊可改变软骨下骨重塑的速率和模式,最终减轻软骨下骨硬化。      

兔实验性骨性关节炎早期软骨下骨板的变化

目的 观察兔膝骨性关节炎软骨下骨板与关节软骨之间联系,同时探讨应用降钙素手段干预软骨下骨板对骨性关节炎中关节软骨保护作用.方法 雄性日本大耳白兔随机分为3组,每组8只.对照组动物暴露右膝关节前交叉韧带而不予切断;模型组与实验组行右膝关节前交叉韧带切断术,术后实验组给予5 IU/(kg·d)降钙素皮下注射,模型组给予等量生理盐水,于术后8周处死所有动物并取材.关节软骨采用HE染色及Mankin评分、Von Gieson染色和基质金属蛋白酶-13免疫组织化学检测,软骨下骨板采用Von Kossa染色方法 进行组织形态计量学分析.结果 软骨组织形态学HE及Von Gieson染色显示,模型组关节软骨明显损伤,Mankin评分结果 显示,模型组评分显著高于对照组和实验组(P<0.05),实验组评分显著高于对照组(P<0.05);免疫组织化学染色及半定量分析表明,模型组基质金属蛋白酶-13表达显著高于对照组和实验组(P<0.05);模型组软骨下骨板形态被破坏,与对照组和实验组比较差异显著(P<0.05).结论 前交叉韧带切断诱导的骨性关节炎兔膝关节软骨与软骨下骨板组织结构同时被破坏,软骨下骨板与关节软骨间存在紧密关系,降钙素早期干预软骨下骨板,能够保护软骨下骨板组织形态结构,对关节软骨起到部分保护作用,延缓骨性关节炎病理进程.     

骨性关节炎对膝关节软骨粘弹性影响实验研究

研究了正常国人新鲜尸体膝关节软骨和骨性关节炎膝关节软骨的粘弹性力学性质.对正常和病态膝关节软骨进行一维拉伸应力松弛、蠕变实验,得出了应力松弛、蠕变实验数据和曲线,以回归分析的方法处理应力松弛、蠕变实验数据,得出了归一化应力松弛函数、归一化蠕变函数表达式.以冯元桢教授的准线性理论得出了正常组和病态组膝关节软骨松弛函数表达式,并对实验结果进行了分析讨论.     

人膝关节软骨下骨成骨细胞和软骨细胞的共培养

背景：软骨细胞移植等组织工程方法已经成为非常有潜力的骨关节炎治疗措施,软骨下骨的成骨细胞对软骨细胞的作用不仅参与骨关节炎的发生和发展,而且与细胞移植治疗预后有着密切的关系。 目的：培养人膝关节软骨下骨成骨细胞和软骨细胞,构建两者共培养的细胞培养模型,探讨软骨下骨成骨细胞对软骨细胞的作用。 方法：收集骨关节炎患者行全膝关节置换和严重创伤患者行截肢后遗弃的胫骨平台组织,采用Ⅰ型胶原酶连续消化后进行贴壁培养的方法,培养软骨下骨的成骨细胞,应用Ⅱ型胶原酶消化方法,培养软骨细胞,然后应用多孔插入器细胞培养池构建成骨细胞和软骨细胞三维共培养模型。 结果与结论：通过免疫组化,NBT/BCIP染色,茜素红染色等检查,证实成功培养出成骨细胞和软骨细胞。实时定量RT-PCR显示,在与骨关节炎的软骨下骨成骨细胞共培养的软骨细胞中,Ⅱ型胶原和蛋白聚糖基因的表达明显下降。可见骨关节炎患者的成骨细胞能促进软骨细胞的退变。     

骨性关节炎对膝关节软骨力学性质影响的实验研究

目的 研究正常国人新鲜尸体膝关节软骨和骨性关节炎膝关节软骨的生物力学性质,为临床提供生物力学参数。方法 取正常和病变的膝关节软骨在ShimadzuAUTOGRAPH电子万能试验机上进行一维拉伸实验。结果 得出了关节软骨最大载荷、伸长比、应力、应变等数据;用最小二乘法处理应力鄄应变数据,得出了其应力鄄应变关系表达式,拟合出了应力鄄应变曲线。结论 病变组各项力学性质指标均明显低于正常组。     

早期骨关节炎软骨下骨趋化因子信号通路的表达

背景：趋化因子广泛存在于炎症反应组织当中,起诱导炎症细胞趋向炎症组织参与免疫应答的作用,大量的实验研究表明,趋化因子在骨关节炎致病过程中起了重要作用。 目的：分析早期骨关节炎软骨下骨的趋化因子信号通路表达改变情况,为阐述其在骨关节炎致病机制中的重要作用提供依据。 方法：30只SD大鼠随机均分为实验组和对照组。实验组切除右膝内侧半月板及内侧副韧带,对照组仅切开关节囊。于造模后1,2,4周取右膝关节标本,采用全基因表达谱芯片技术研究软骨下骨全基因表达,利用差异基因分析、信号通路分析的方法分析趋化因子信号通路的表达改变情况。 结果与结论：①发现一系列与趋化因子信号通路相关的差异表达基因。②实验组与对照组相比较,趋化因子信号通路在造模后1周差异有显著性意义(P < 0.05),造模后2,4周差异无显著性意义(P > 0.05)。说明趋化因子信号通路在极早期膝关节骨关节炎的软骨下骨改变中起重要作用。     

Ultrastructural change of the subchondral bone increases the severity of cartilage damage in osteoporotic osteoarthritis of the knee in rabbits

Osteoporotic osteoarthritis is a phenotype of osteoarthritis (OA) manifested as fragile and osteoporotic subchondral bone. However, the ultrastructural features of subchondral bone in osteoporosis OA have not been determined. The study was aimed to investigate the ultrastructural dynamic changes of subchondral bone in osteoporotic OA model and how the ultrastructural damage in the subchondral bone caused by osteoporosis deteriorated the cartilage damage in OA. Eighteen rabbits were equally randomized to three groups, including the control, the OA and the osteoporotic OA groups. The structural changes of cartilage were evaluated by HE and safranin-O fast green staining, the Mankin’s grading system was used to assess the stage of OA progression. And microstructural or ultrastructural changes in subchondral bone were assessed by micro-computed tomography or by scanning electron microscopy. According to the changes of cartilage histopathology, the OA group was in the early pathological stage of OA while the osteoporotic OA group was in the middle stage of OA based on Mankin’s grading system. In addition, the damage of cartilage surface, reduction in the number of chondrocytes and the matrix staining were more increased in the osteoporotic OA group compared to the OA group. Compared to the OA group, the subchondral bone in the microstructure and ultrastructure in the osteoporotic OA group showed more microfracture changes in trabecular bone with more destructions of the tree-like mesh. Moreover, the collagen fibers were random rough with a fewer amount of bone lacunae in subchondral cortical plate in the osteoporotic OA group compared to the OA group. These findings indicated that the subchondral bone ultrastructure in the osteoporotic OA model was characterized by the destruction of the network structure and collagen fibers. The subchondral bone ultrastructural damage caused by osteoporosis may change mechanical properties of the upper cartilage and aggravate OA cartilage. Therefore, early diagnosis and treatment of osteoporosis is of great significance to prevent early OA from further developing osteoporotic OA.     

MMP3在早期骨关节炎模型软骨下骨的表达及其意义

目的: 研究早期骨关节炎软骨下骨基质金属蛋白酶3(MMP3)的表达情况,为阐述其在骨关节炎致病中的重要作用提供依据。方法: 大鼠分为实验组(n=30)和对照组(n=30)。实验组切除右膝内侧半月板并切断内侧副韧带,对照组仅切开关节囊。于术后1、2、4周取右膝关节标本,病理检查验证造模成功后,采用qPCR技术及免疫组化技术从基因水平及蛋白水平全面分析软骨下骨MMP3的表达情况。结果: (1)术后1周,软骨下骨的MMP3在实验组的表达量明显升高,是对照组的8.34倍(P＜0.05);术后2周,MMP3在实验组的表达量也升高,是对照组的4.85倍(P＜0.05),但相对于术后1周,表达量有所下降;术后4周,2组的MMP3表达量无显著差异;(2)术后1周与术后2周,实验组软骨下骨区检测到大量的MMP3阳性细胞,以小单核细胞及多核巨细胞为主,术后4周实验组、对照组及阴性对照组织切片均未检测到MMP3阳性细胞。结论: 软骨下骨MMP3表达在早期骨关节炎致病机制中起了重要作用。     

关节软骨下骨血管新生的研究进展

骨关节病是一种很常见的疾病,其导致的疼痛及功能障碍对人体健康及生活质量有很大影响,但其发病原因及病理机制尚未完全清楚。近年来,骨关节病患者的软骨下骨改变及血管新生备受重视。本文就骨关节病与软骨下骨血管新生的关系及其研究进展作一综述。     

软骨下骨量变化与软骨退变的相关性

背景：关节软骨与软骨下骨在骨关节炎病变进程中的相互作用机制目前尚未完全阐明。软骨下骨量改变在骨关节炎病理进程中亦发挥重要作用。 目的：分析2种手术方式及2种蛋白酶诱导建立兔膝关节骨关节炎动物模型的效果,以及软骨下骨量变化与关节软骨退变的相关性。 方法：32只新西兰大白兔随机分为4组：Hulth模型组、前交叉韧带切断模型组、Ⅱ型胶原蛋白酶组及木瓜蛋白酶组,每组8只,右膝关节造模,左膝关节作为自身对照。造模后0,4,8周行DXA扫描,8周行MRI扫描后处死实验动物,取双侧膝关节制作病理组织学切片,比较各组膝关节影像学表现、大体形态及病理变化,并采用Mankin评分进行定量分析。 结果与结论：造模后0,4,8周实验侧膝关节骨密度进行性降低,骨量降低程度Hulth模型组>前交叉韧带切断模型组>Ⅱ型胶原蛋白酶组>木瓜蛋白酶组。MRI显示实验侧股骨内外髁关节软骨厚度变薄,厚度Hulth模型组<前交叉韧带切断模型组<Ⅱ型胶原蛋白酶组<木瓜蛋白酶组。大体标本、组织切片观察及Mankin评分显示手术建模组骨关节炎程度较药物组重,Hulth模型组病变最重,木瓜蛋白酶组最轻。结果说明关节内手术及关节腔内注射蛋白酶均能建立骨关节炎动物模型;手术造模可复制出中晚期骨关节炎,药物诱导可产生骨关节炎早期改变。骨关节炎病变严重程度与软骨下骨骨密度呈负相关;关节软骨退变和软骨下骨改变相互关联,病变进行性发展。     

Subchondral bone changes and chondrogenic capacity of progenitor cells from subchondral bone in the collagenase-induced temporomandibular joints osteoarthritis rabbit model

Purpose: The goals of this study were to characterize subchondral bone changes, and to determine biological activity characteristics of progenitor cell populations from subchondral bone in the collagenase-induced temporomandibular joint osteoarthritis (TMJOA) rabbit model. Greater understanding of such pathological changes occurring in TMJOA samples is critical in the future treatment modalities regarding cartilage protection and repair. Furthermore, the use of progenitor cell populations in various cartilage regeneration strategies proves to be a fruitful avenue for research and clinical applications. Materials and methods: Bone remodeling and anabolic activity of subchondral bone was evaluated by hematoxylin-eosin (H&E), Alcian blue-periodic acid-Schiff (AB-PAS) staining and immuohistochemical staining. The biological activity characteristics of progenitor cells were assessed by expressions of collagen type II, CD44, SOX-9 and MMP-9 by immunohistochemistry and Western blot analysis. Results: In most of the specimens, cartilage of the digested area displayed a reaction characterized by thickening of the cartilage cellular structure with retraction structure formation in the subchondral bone. Most of the specimens focuses on chondroid metaplasia were observed in the subchondral bone, promoting its remodeling, which could develop to endochondral ossification and increasing subchondral bone size. Meanwhile, immunohistochemistry analysis revealed that CD44 expressions in subchondral bone were most significantly increased in TMJOA at 2 weeks group (P < 0.01). And, at 4, 6 and 8 weeks groups, the osteochondral junction had completely disappeared by active subchondral bone remodeling, and collagen type II, CD44, SOX-9 and MMP-9 expressions in active subchondral bone region were significantly increased in TMJOA (P < 0.05). In addition, western blot analysis revealed that CD44 expression significantly emerged in subchondral bone region at 2 weeks group (P < 0.01). Meanwhile, SOX-9 expression emerged in all group, and the intensity was increased in the experimental groups (P < 0.05). Conclusion: Our results suggest that the beneficial activation of progenitor cells and bone marrow stem cells in subchondral bone at early stage of TMJOA played an important role on renovation and remodeling of subchondral bone.     

兔膝骨性关节炎滑膜细胞体外培养和鉴定

目的 探讨复制膝骨性关节炎的实验动物模型;探讨兔膝关节滑膜细胞体外分离、培养的方法及生长特性.方法 改良Hulth法复制膝骨性关节炎,于造模后4周机械分离滑膜组织;用改良酶消化法分离滑膜细胞进行培养,观察滑膜细胞的生长情况.结果 改良Hulth法复制膝骨性关节炎模型,滑膜增生明显;酶消化法分离培养获得大量滑膜细胞且细胞...     

Rapid decalcification of articular cartilage and subchondral bone using an ultrasonic cleaner with EDTA

Articular cartilage and subchondral bones were used to be the samples for studying effects of drugs in the joint degenerative diseases such as osteoarthritis. Because of the deposition of mineral salts, articular cartilage and subchondral bones require decalcification process to soften the tissues. EDTA is a chelating agent that is commonly used to remove mineral salts, but this step is time-consuming and can take as long as 45 days. Commercial ultrasonic cleaner and microwave oven were reported to reduce the decalcification timing. The aim of this study is to determine and compare the decalcification of human articular cartilage and subchondral bone using EDTA together with ultrasonic cleaner or microwave oven. Hundred pieces of articular cartilage and subchondral bones obtained from osteoarthritis patients undergone total-knee-replacement were divided into 10 groups according to decalcification method (ultrasonic cleaner or microwave) and timing (2, 4, 6, 8, and 10 h). In each group, all cartilage and subchondral bone pieces were decalcified and sectioned, and subsequently stained with haematoxylin and eosin, Von Kossa, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, or caspase-3 immunohistochemistry. The optimal timing of decalcification of articular cartilage and subchondral bones using EDTA together with ultrasonic cleaner was at 8 and 10 h, while the timing using EDTA together with microwave oven was more than 10 h. Clear TUNEL and caspase-3 signals were obtained from samples decalcified using EDTA together with ultrasonic cleaner for 8 h. In summary, to our knowledge, this is the first study that compared EDTA decalcification between ultrasonic cleaner and microwave oven. Here, we report a new methodology for decalcification for articular cartilage and subchondral bones that reduces decalcification time from weeks to hours and is suitable for further pathological analyses.     

The subchondral bone plate of the femoral head in adult rabbits

Summary The mineralized structures and spontaneous remodelling of the subcondral bone-plate in the femoral head of adult rabbits of different ages were studied by microradiography and fluorescence microscopy after administration of Tetracycline between 8 months and 2 days before death. In animals over 1 year of age the subchondral bone-plate was constructed of evenly mineralized osteones, located mainly in the immediate vicinity of the calcified articular cartilage and lamellar bone bordering on the medullary cavity. In some small areas lamellar bone was the only bone tissue separating the medullary cavity from the calcified articular cartilage. In adult animals between 8 and 10 months of age the subchondral boneplate was absent in minor areas, chiefly in the lateral parts of the femoral head. In the fluorescence microscopic study in animals older than 1 year (a) the osteones in the subchondral bone showed a very low labelling frequency, (b) fluorescence was observed mainly on the lamellar bone surfaces bordering on the medullary cavity and (c) the labelling in the cortical part of the subchondral bone was still present up to 8 months after administration of the Tetracycline. The cancellous bone of the epiphysis showed a low labelling frequency which was of the same order as that in the lamellar surfaces belonging morphologically to the subchondral bone plate.From these observations it was concluded that (1) the cortical part of the subchondral bone-plate in rabbits over 1 year of age undergoes a low degree of spontaneous remodelling and (2) the cancellous bone of the epiphysis and the lamellar bone belonging morphologically to the subchondral bone-plate undergo low but similarly spontaneous remodelling. Duplicate fluorescence of the tidemark observed in a number of animals was considered to indicate that the mineralization of the articular cartilage with increasing age slowly progresses towards the articular surface.