关节软骨力学特征研究进展

骨关节炎(osteoarthritis OA)是一种以关节疼痛和僵硬为特征的慢性退行性关节疾患,好发于老年人群。OA发病缓慢,病程较长,早期临床表现和组织学改变均不明显,限制了疾病的早期诊断与治疗。关节软骨微观结构决定了软骨宏观力学特性。软骨微观结构具有区域差异性,导致软骨的力学性能也具有区域依赖性,从软骨浅表区到深区软骨抗负荷、抗形变能力逐渐增加。然而,在OA病程发展过程中,软骨微观构成改变导致OA软骨抗负荷、抗形变能力降低。通过检测关节软骨的微观构成可以推测软骨的力学特性,反之检测软骨的力学指标可以了解软骨早期的微观改变,从而有助于了解OA的病程发展,便于疾病的早期诊断。综述近年来关节软骨在正常和急慢性损伤状态下力学性能的相关研究文献,阐述软骨结构与力学性能之间的关系,为OA的病程发展、早期诊断与治疗提供进一步理论依据。     

关节软骨生物力学与力学生物学 2022 年研究进展

关节软骨是动关节内骨表面具有弹性的负重结缔组织,能提供低磨损润滑、缓冲震荡、传递载荷等支撑作用,具有层级纤维复合结构和优异的力学性能。软骨内没有血管、神经和淋巴,代谢缓慢,损伤后难以实现自我修复。目前,高发的关节炎疾病仍是基础与临床研究的一大热点。关节软骨是力学敏感组织,力学环境影响着组织不同方向的发展。2022年,学者们继续对关节软骨的生物力学与力学生物学开展大量研究;对软骨形态、功能与力学状态,以及不同条件下软骨力学状态的研究报道较多;研究设计了一些软骨相关的动物、组织及细胞水平的加载装置,也开展了体外及在体力学载荷下软骨退变、损伤的修复研究,获得了重要的修复方法及手段。关节软骨的生物力学与力学生物学研究是关节炎、软骨缺损及修复的基础,关节软骨损伤修复定量力学条件的影响还需体内和体外深入研究。     

关节软骨体外构建力学环境的研究进展

很多物理因素都影响软骨组织的生长和发育,其中力学因素起主要作用。软骨的生长、发育是力学调控的适应过程。当前采用多种力学条件应用于软骨生物反应器,如流体剪应力、液体压力、直接压缩等,或其中部分组合,但这些条件还没有构建出与活体软骨结构-功能相匹配的人工软骨。如果一种载荷能适合构建软骨,那么这种载荷首先能保证培养物内部信号分子、营养和废物的有效运输;其次,能对支架内种子细胞特定的力学刺激;第三,能促进培养物结构-功能的发展。本文回顾、分析当前多种力学条件的作用效果,其中流体剪应力、液体压力、拉伸、直接压缩或变形剪应力都是软骨受力状态的部分体现。作者认为滚压载荷是软骨培养的合适力学环境,它是当前多种力学条件的一个综合指标,对软骨培养物可以形成纵向的动态压缩和横向的动态变形剪应力,并且有利于细胞新陈代谢物质的运输,因此,滚压环境可能是人工软骨结构-功能构建的发展方向。     

模拟微重力对大鼠关节软骨力学特性的影响

目的基于大鼠尾吊实验,研究模拟微重力对关节软骨组织力学特性的影响。方法通过14 d的尾吊实验建立模拟微重力作用的大鼠模型,获得12块后肢大腿骨关节处软骨样本,运用高频超声测量技术获取大鼠关节软骨特定位置因自由膨胀引起的组织应变,基于三相模型估计软骨组织的轴向弹性模量,并比较尾吊组和对照组大鼠关节软骨的力学特性。结果尾吊组大鼠后肢特定测量位置处软骨厚度的平均值略小于对照组,但差异不显著;尾吊组大鼠特定位置关节软骨组织的平均轴向弹性模量为(5.05±2.98)MPa,小于对照组((6.31±3.37)MPa),两者存在着显著差异(P<0.05)。结论微重力环境会影响关节软骨组织的力学特性,本研究结果为人类长期的太空活动提供参考信息。     

关节软骨胶原纤维增强特性

目的在COMSOL关节软骨计算中引入胶原纤维增强特性,并分析引入前后模型结果的差异。方法以软骨中胶原纤维的走向为基础,对胶原纤维的应力单独建模,将胶原纤维的应力写入到原来软骨多孔弹性模型中。为避免应变2次项的出现采用了函数调用方式,同时提高求解器的迭代次数以更好收敛。结果胶原纤维增强模型表面的Y方向初始位移仅为15μm,是非增强模型的17.6%。增强模型的X方向正应变仅为非增强模型的10%,而近表面的X方向正应力超过非增强模型的10倍。结论在COMSOL关节软骨计算中实现了胶原纤维增强特性的引入,为软骨胶原纤维损伤提供了计算模型和理论分析。胶原纤维通过横向增强约束了竖直方向的应变,提高了软骨承载能力,改善了软骨的力学性能。     

不同病理学等级关节炎软骨的三相力学特性

目的基于超声膨胀观测技术与三相理论提取关节软骨的轴向弹性模量,探索不同等级关节炎软骨的三相力学特性。方法对不同阶段的兔子关节炎软骨样本进行病理学分级,基于高频瞬态超声测量技术获取软骨因自由膨胀引起的组织应变量,结合组织的固定电荷密度和水体积分数,运用三相模型估计软骨的轴向弹性模量,并进行相关性分析。结果正常软骨与关节炎软骨的轴向弹性模量存在着明显差异(P<0.05),不同等级软骨样本的轴向弹性模量存在明显差异(P<0.05)。正常软骨组织的平均弹性模量为(15.87±6.30)MPa,随着关节炎的发生与关节炎程度的加重,软骨的弹性模量逐渐减小,1、2、3级软骨样本的轴向弹性模量分别为(11.33±5.21)、(9.15±5.68)、(6.05±4.99)MPa。结论不同病理学关节炎等级软骨样本的三相力学特性有明显差异,该研究为应用软骨组织的力学属性定量评判关节炎的严重程度提供了新的思路。     

关节软骨的微摩擦接触力学特性

目的使用修饰后的原子力显微镜(atomic force microscopy,AFM)探针研究关节软骨的微摩擦接触力学性能。方法使用微操作器对AFM氮化硅探针进行修饰处理,具体操作为在探针上粘贴玻璃微球作为针尖,然后使用修饰后的探针研究人体和牛关节软骨的微摩擦接触力学性能。结果人体和牛软骨的粗糙度分别为(68.63±6.22)、(50.16±6.47)nm,随着载荷的增大,人体和牛软骨的摩擦力逐渐增大。当探针滑动速度从0增加到100μm/s时,试样与探针之间的摩擦力增速很快;当速度从100μm/s增加到300μm/s时,摩擦力上升缓慢。结论软骨表面具有明显的纤维状结构,软骨的粗糙度与测量范围直接相关。随着速度或载荷的增大,人体和牛软骨的摩擦力增大,变化范围相同。探讨关节软骨在微摩擦试验中的力学和摩擦学性能表现,对于认识软骨损伤机制和医用人工关节抗磨材料的开发具有重要意义。     

关节软骨组织工程的研究进展

介绍了关节软骨组织工程中种子细胞、支架材料的研究现状以及生长因子在关节软骨组织工程中的应用进展，阐明了随着对细胞行为、支架材料特性、细胞与材料的组合构建研究的深入，组织工程在关节软骨修复方面应用前景十分广阔。     

T_2 mapping评价关节软骨损伤及修复的研究进展

T2mapping是近年来发展的MRI新技术,能间接反应关节软骨结构及组织学成分的变化,发现关节软骨形态学变化之前的组织学改变。笔者就T2mapping成像原理、软骨损伤的组织学表现,以及成像影响因素等方面作一综述,重点介绍了T2mapping在软骨损伤、退变、修复等方面的研究进展,并展望了T2mapping未来发展前景。     

关节软骨压缩特性的实验研究

目的 探讨正常关节软骨的压缩特性。方法 将人的股骨头软骨制成圆柱形标本,分A、B、C、D四组。应用两种不同力学实验装置分别对A、B组标本加载,测定标本在受压后1秒末的应力和应变值,作出应力-应变曲线图。C组标本在恒定压力下受载,测量标本在受压后不同时间应变值的变化。D组标本受压后并保持一定的应变值,观察关节软骨受压后不同时间压力的变化。结果(1)正常关节软骨的瞬时应力-应变曲线呈非线性关系,应力越大,弹性模量值越高;两种装置的实验结果具有明显差异性。(2)关节软骨在恒定应力作用下,应变随时间的延长而增大,     

胶原纤维束对软骨力学性能的影响

目的 研究胶原纤维束对软骨力学性能的影响,为临床医生指导早期软骨损伤患者的康复运动提供参考。方法 建立一种纤维增强的多孔黏弹性二维数值模型,考虑纤维分布、弹性模量、孔隙率和渗透率随软骨深度的变化关系。研究纤维束局部断裂和从表面渐进断裂以及纤维束尺寸对软骨力学性能的影响,获得软骨基质的最大主应变。结果 基质的最大主应变出现在软骨中层靠上某个位置,此位置不受纤维断裂模式和纤维束尺寸的影响。含较粗纤维束软骨的应变降低。结论 软骨中层易发生力学损伤,纤维束增粗可以降低基质的最大主应变,一旦纤维束发生断裂,较粗纤维束的软骨的基质最大主应变更大,使软骨更易发生损伤演化情况。     

SOX Trio Decrease in the Articular Cartilage with the Advancement of Osteoarthritis

SOX trio (SOX-5, SOX-6, and SOX-9) maintain the chondrocytic phenotypes and are vital for chondrogenesis in embryonic development. The purpose of this study is to investigate the change in the expression of SOX trio with the advancement of osteoarthritis (OA) in human articular cartilage (AC). Human OA samples from eight patients were obtained from the distal femoral condyles during total knee arthroplasty. Minimally OA cartilage taken from areas with no obvious surface defects on lateral condyles was compared with advanced OA cartilage obtained from areas within 1 cm of overt lesion located on medial condyle surface. SOX-5, SOX-6, and SOX-9 gene expressions significantly decreased by 41% (p = 0.047), 46% (p = 0.047), and 56% (p = 0.029) in advanced OA area compared with the minimally OA area. There was a significant decrease in aggrecan and type II collagen (COL2A1) gene expressions by 73% (p = 0.029) and 65% (p = 0.029), respectively, in advanced OA area compared with the minimally OA area. From Western blotting and immunohistochemistry, SOX-5, SOX-6, SOX-9, type II collagen, and aggrecan protein expressions also significantly decreased in advanced OA cartilage compared with minimally OA cartilage. DNA methylation study of SOX-9 promoter regions revealed no difference in the epigenetic status between the two areas. It is concluded that SOX trio gene and protein decreased with advancement of OA in human articular cartilage.     

高频超声对关节软骨消肿与肿胀行为的定量分析

关节软骨可减轻关节滑动中所产生的关节面间的摩擦,具有负重传导、润滑及耐磨损的特性。关节软骨肿胀属性的改变可作为检测早期关节炎的指标之一。本研究的目的在于利用高频超声技术和信号处理技术建立一个实时超声监测系统,以一种非接触、无损的方法研究软骨的瞬时渗透性消肿(收缩)及肿胀(膨胀)行为。样本取自新鲜成年牛膝盖骨,通过改变外溶液的浓度引入渗透压,促使软骨发生收缩膨胀反应,利用实时超声监测系统进行无损测量。实验表明关节软骨的肿胀消肿过程出现“过激松弛”现象,最大收缩膨胀应变的绝对值分别为0.75%±0.57%和0.28%±0.28%,膨胀应变与收缩应变具有显著性差异(p<0.05)。同时发现软骨内超声速度随时间的指数性变化可以表征离子的渗透过程。实验证明该超声系统具有良好的可重复性(组内相关系数ICC>0.98)和可靠性(相对误差<5%)。应用高频超声技术不仅可无损量化地研究软骨的渗透性消肿膨胀行为,而且为今后研究软骨病变及早期发现关节炎提供一个有效可行的方法。     

老龄关节软骨和软骨下骨的变化及相互作用机制研究进展

衰老是骨关节炎(osteoarthritis,OA)和骨质疏松等疾病的主要影响因素。然而,它们并不是衰老的必然结果,而且与衰老有关的骨和软骨的变化与疾病发展之间的关系尚不清楚。研究表明,骨关节炎的发生和发展并不是简单的软骨磨损过程,它的发生包括整个关节各组织复杂的生物、化学和力学变化,特别是软骨和软骨下骨之间生化和力学的相互作用。衰老促成OA的发生和发展,但并不是直接导致OA的原因。与衰老相关的变化为OA的发生提供了一个基础,使关节更容易受到其他因素(如异常的生物力学和生物化学)影响,从而促进OA发展。因此,了解衰老影响关节组织的基本机制可能会为减缓或预防OA发展提供新的靶点。从衰老过程中软骨和软骨下骨的年龄相关性变化、力学传导作用和血管新生机制3个方面对相关研究进展进行综述。     

低温保护剂渗透关节软骨研究进展

玻璃化保存关节软骨具有临床治疗和科学研究的双重意义。如何在保证对软骨细胞造成的损伤最小的前提下,将足够多的低温保护剂载入软骨组织,是玻璃化保存关节软骨的关键环节之一,而低温保护剂渗透关节软骨的特性是指导设计低温保护剂加载处理过程的重要依据。从实验研究和数学建模两方面,系统评述低温保护剂渗透关节软骨的研究进展,指出现有研究存在的不足,并对今后低温保护剂渗透关节软骨的研究方向提出展望。     

低温保护剂和降温速率对猪关节软骨压缩杨氏模量的影响

利用动态热机械分析仪(DMA),研究了3种低温保护剂(DMSO、甘油、丙二醇)、4种不同低温保护剂浓度(10%V/V、30%V/V、60%V/V、80%V/V)以及两种降温速率(1℃/min和20℃/min)对猪关节软骨杨氏模量的影响。研究结果表明:在相同的低温保护剂浓度条件下,样品经DMSO处理过的杨氏模量最大,甘油的次之,丙二醇的最小,这是由3种保护剂官能团的水合作用所决定的;随着低温保护剂浓度的增加,杨氏模量也增加,如降温速率为20℃/min时,10%甘油的杨氏模量为1.68 MPa,30%甘油的杨氏模量为2.15 MPa,60%甘油的杨氏模量为2.97 MPa,其它两种低温保护剂也是呈同样的趋势;较慢的降温速率更有利于关节软骨生物力学特性的保存,如新鲜关节软骨的杨氏模量为2.53 MPa,当丙二醇浓度为60%、降温速率为1℃/min时的杨氏模量为2.38 MPa,20℃/min时的杨氏模量为2.25 MPa,其他两种低温保护剂也呈同样的趋势。     

Quantification of Stiffness Change in Degenerated Articular Cartilage Using Optical Coherence Tomography-Based Air-Jet Indentation

Articular cartilage is a thin complex tissue that covers the bony ends of joints. Changes in the composition and structure of articular cartilage will cause degeneration, which may further lead to osteoarthritis. Decreased stiffness is one of the earliest symptoms of cartilage degeneration and also represents the imperfect quality of repaired cartilage. An optical coherence tomography (OCT)-based air-jet indentation system was recently developed in our group to measure the mechanical properties of soft tissues. In this study, this system was applied to quantify the change of mechanical properties of articular cartilage after degeneration induced by enzymatic digestions. Forty osteochondral disks (n = 20 × 2) were prepared from bovine patellae and treated with collagenase and trypsin digestions, respectively. The apparent stiffness of the cartilage was measured by the OCT-based air-jet indentation system before and after the degeneration. The results were also compared with those from a rigid contact mechanical indentation and an ultrasound water-jet indentation. Through the air-jet indentation, it was found that the articular cartilage stiffness dropped significantly by 84% (p < 0.001) and 63% (p < 0.001) on average after collagenase and trypsin digestions, respectively. The stiffness measured by the air-jet indentation system was highly correlated (R > 0.8, p < 0.001) with that from the other two indentation methods. This study demonstrated that the OCT-based air-jet indentation can be a useful tool to quantitatively assess the mechanical properties of articular cartilage, and this encourages us to further develop a miniaturized probe suitable for arthroscopic applications.     

Age-Related Changes in the Synthesis of Matrix Macromolecules by Bovine Articular Cartilage

Calf and mature cow articular cartilage was labeled in vitro with [³⁵S]SO₄ and [³H]glycine and kinetics of incorporation of both isotopes by cartilage fragments was determined by scintillation spectroscopy. The cartilage fragments were then extracted in sequence with 4M GuHCl (Guanidium chloride) and pepsin. The pepsin digest was adjusted to 1.3 M NaCl and pepsin-solubilized collagen salted out. The 4M GuHCl extract, collagen and pepsin-resistent residue were then freeze-dried. The 4M GuHCl extract was further fractionated by DEAE (Diethylaminoethyl) 52 ion exchange chromatography to obtain protein and PG (Proteoglycan) fractions. The protein fraction was also characterised by SDS-PAGE and PG fraction by Sepharose C1–2B chromatography under associative conditions in the presence and absence of an exogenous HA (Hyaluronic acid). The GAG (Glycosaminoglycan) side chains of the PG samples were analysed by Sephadex G-200 column chromatography and their composition determined by paper chromatography after chondroitinase ABC digestion.

Linear incorporation of both isotopes was observed from 1 to 18 hours of incubation and roughly equal amounts of f³⁵S]SO₄ counts were found on per cell bases in both cartilages although less [³H]glycine was incorporated by cow chondrocytes. It was also found that calf chondrocytes synthesize much greater proportion of the collagen whereas the cow cells synthesize PGs of smaller hydrodynamic sizes, bearing shorter GAG side chains that are enriched in KS (Keratan sulfate) and Ch-6S (Chondroitin-6 sulfate isomer). A failure of cow ³⁵S-PGs monomers to interact with an exogenous HA in the presence of other extracted components was also demonstrated.

The relevance of these findings for the mechanism of cartilage damage in aging and osteoarthritis is discussed.     

Irreversible Changes in Glycosaminoglycan Composition of Anatomically Intact Bovine Articular Cartilage Induced by Intermittent Loading

The changes in matrix composition induced by 1 MPa intermittent (0.2 Hz) loading of anatomically intact bovine articular cartilage in vitro are studied. The kinetics of chondrocyte response was determined in experiments where sesamoid bones of adult cows were loaded for 0, 3, 5 and 7 days. The reversibility of the induced changes were studied in sesamoid bones that were loaded for 5 days and subsequently cultured without loading for another 1, 2 and 3 weeks. Water content was not affected by loading, nor by subsequent unloaded culture. Glycosaminoglycan content was not affected by loading, nor by subsequent unloaded culture for another 2 weeks. However, after 3 weeks of culture following 5 days of loading, a significant decrease in glycosaminoglycan content was found. Glycosaminoglycan synthesis was already increased after 3 days of loading. Loading for longer periods (5 and 7 days) further increased the glycosaminoglycan synthesis rate. Glycosaminoglycan synthesis decreased during the first week of subsequent culture without loading. In the third week it dropped to a lower level than observed for the control. The amount of 3-B-3(-) epitope was increased and the length of newly synthesized glycosaminoglycans was decreased by intermittent loading. Subsequent unloaded culture did not further increase the amount of 3-B-3(-) epitope. However, after 3 weeks of unloaded culture the newly synthesized glycosaminoglycans were as long as those synthesized in the control. The results suggest that the changes in glycosaminoglycan chain length were reversible. However, the overall chondrocyte response to our loading regime seems to be detrimental to the tissue: expression of 3-B-3(-) epitope induced by loading together with the drop in glycosaminoglycan content and synthesis observed at the end of the total culture period indicate that the cartilage is irreversibly damaged.