力学刺激促进骨髓基质干细胞体外软骨分化

目的探讨离心力刺激对猪骨髓基质干细胞(BMSCs)体外成软骨分化的作用,以及对三维支架上构建组织工程化软骨的影响,明确力学刺激与细胞分化及组织形成的关系,为体外软骨构建提供适当参数。方法抽取8周龄猪髂嵴骨髓,应用贴壁法分选单个核细胞,体外培养扩增后获得第2代BMSCs,以5.0×107/cm3的细胞密度接种到聚羟基乙酸(PGA)制成的圆柱形三维支架上,7d后分成4组,在不同的力学条件及诱导条件下培养。8周后取材,行相关检测。结果力学诱导组形成的组织呈白色,细腻光泽,形状规则,体积无明显改变,有良好的弹性和硬度,并具有典型的软骨陷窝结构和大量软骨特异性细胞外基质,Ⅱ型胶原及丰富的聚合蛋白多糖(GAG)成分。GAG含量为(6.0±1.2)mg/g,抗压强度为(2.2±0.8)kPa,弹性模量为(7.4±1.6)kPa。各项指标均明显优于其他各组。结论力学刺激有利于促进BMSCs成软骨分化,并在三维支架材料上构建组织工程化软骨。     

支架及无支架条件下构建组织工程软骨的研究进展

支架材料的研究是组织工程的研究热点之一,软骨细胞复合培养给组织工程软骨的构建带来希望,单一的材料诸如胶原、透明质酸、壳聚糖、纤维蛋白凝胶等已经证明可以与软骨细胞复合培养,两种或者多种材料复合可以提高材料的性能,更好地用于组织工程软骨的构建,并满足软骨缺损修复的需要;同样,在无支架情况下应用软骨细胞聚集培养、沉淀培养的方法,可以构建组织工程软骨,并给软骨缺损的修复带来新希望,但目前的研究较少。两者是组织工程软骨构建的两个主要方向。     

自体骨膜游离移植的实验研究--不同龄动物大块关节软骨缺损修复的比较

目的研究比较自体骨膜移植软骨再生修复不同龄动物大块关节软骨缺损。方法用52只不同龄家兔自体骨膜游离移植修复大块关节软骨缺损,比较移植骨膜生发层朝向关节腔与松质骨时再生软骨的差别。结果经不同时期肉眼和组织学检查证实,幼年兔和成年兔的骨膜移植都能生成软骨,修复大块关节软骨缺损。在成年兔骨膜再生的软骨与成年兔本身周围正常软骨的厚度、组织结构一样。移植骨膜生发层朝向关节腔与松质骨二者间再生软骨结果无明显差别。结论骨膜具有再生软骨的能力,可用来移植修复关节软骨的缺损。骨膜移植生发层不同朝向对软骨再生无明显影响。成年后骨膜移植修复关节软骨缺损能够生成与自身相适应的软骨。     

Histological comparison of patellar cartilage degeneration between chondromalacia in youth and osteoarthritis in aging

The histological findings of the patellar cartilage were compared between cases of chondromalacia, which occurs predominantly in young persons (22 patients, average age 19.8 years) and cases of osteoarthritis, which is common among the elderly (21 patients, average age 65.4 years). The histological findings of cartilage in the chondromalacia were characterized by increased density and vigorous fibrous metaplasia of chondrocytes. These findings may be considered to represent a reactive change in the chondrocyte. Cartilage degeneration in osteoarthritis, by contrast, is regressive and presents a clearly different histological picture from that of chondromalacia patellae. We conclude that chondromalacia does not easily lead to osteoarthritis. On the other hand, the cartilage was characteristically softened, as observed by gross inspection, and showed rarefaction of the cartilage matrix. It should be noted that the change was not observed in aging, but showed a pattern of cartilage degeneration peculiar to young patients with chondromalacia patellae.     

Chondrocyte-seeded hydroxyapatite for repair of large articular cartilage defects. A pilot study in the goat

The aim of this study was to evaluate the potential for restoration of a large cartilage defect in the goat knee with hydroxyapatite (HA) loaded with chondrocytes. Isolated chondrocytes were suspended in fibrin glue, seeded on top of the HA, and then the composite graft was implanted in the defect. After transplantation, cell behaviour, newly synthesised matrix and the HA–glue interface were assessed histologically after 2, 4, 12, 26 and 52 weeks. Special attention was paid to the incorporation process of HA in the subchondral bone and interactions between this biomaterial and the fibrin-glue–chondrocyte suspension.

Chondrocytes in the glue proved to survive the transplantation procedure and produced new metachromatically stained matrix two weeks after implantation. The glue–cell suspension had penetrated the superficial porous structure of the HA. Four weeks after surgery, islands of hyaline-like cartilage were observed at the HA–glue interface. A layer of fibrous tissue was formed surrounding the HA graft, resulting in a relatively instable fixation of the HA in the defect. This instability of the graft in the defect, possibly together with early weight bearing, resulted in a gradual loss of the newly formed hyaline cartilage-like repair tissue. Progressive resorption of the HA occurred without any sign of active bone remodelling from the host site. One year after surgery part of the defect which extended down to the cancellous bone had been predominantly restored with newly formed lamellar bone. Only small HA remnants were still present at the bottom of the original defect. Resurfacing of the joint had occurred with fibrocartilaginous repair tissue.

The absence of adequate fixation capacity of the HA near the joint space resulted in a relative instability of the graft with progressive resorption. Therefore, HA is not a suitable biomaterial to facilitate the repair of large articular cartilage defects.     

游离骨膜移植修复关节软骨缺损的实验研究

在家兔的股骨内髁上作一关节软骨全层缺损,且从同侧胫骨近端取下一片骨膜骨块,将骨膜生发层向外包绕骨块,嵌入缺损处,观察其作用。实验表明,骨膜在一定条件下可以化生成软骨;滑液环境与关节运动在软骨化生过程中起重要作用;透明软骨化生和再生率与运动量及开始时间相关;自体骨可作为一种将游离骨膜固定在软骨缺损内的填充材料。     

鹿茸多肽诱导自体骨髓间质干细胞移植修复兔膝关节软骨缺损

目的探讨经鹿茸多肽(PAP)诱导的兔自体骨髓间质干细胞(MSCs)复合胶原膜(MCMG)对兔膝关节局部全层软骨缺损的修复作用。方法新西兰大白兔随机分成A,B,C3组,A组以PAP诱导后的自体MSCs和MCMG修复软骨缺损;B组以经转化生长因子β1(TGF-β1)诱导后的自体MSCs和MCMG修复软骨缺损;C组以自体MSCs和MCMG修复软骨缺损。分别于术后2、4和8周取材进行大体、组织学及免疫组织化学染色观察,根据关节软骨组织学计分标准进行评分。结果对术后8周大体及各阶段组织学形态评分结果显示:A组与B组修复差别无统计学意义(P>0.05),而A、B组明显优于C组(P<0.05)。结论经PAP诱导的兔自体MSCs/MCMG支架复合物可促进软骨缺损的快速修复,恢复软骨组织的结构和功能。     

细胞因子与骨性关节炎软骨退变的研究现状

骨性关节炎是最常见关节疾患之一,是导致中老年残疾的最常见原因,严重危害着中老年人的生活质量。骨性关节炎的主要病理是软骨的改变,软骨细胞分解和合成活动的平衡有助于维持软骨细胞外基质结构和功能的完整性,而软骨细胞的这种平衡受到细胞因子制约,本文就近年来细胞因子与OA软骨退变之间的研究现状综述。