无滑膜肌腱滑膜化的实验研究

实验将胎儿指屈肌腱鞘内滑膜组织进行体外培养，经原代培养和传代培养制成滑膜细胞悬液，再与胎儿掌长肌肌腱联合培养。结果显示，滑膜细胞爬行并覆盖了掌长肌肌腱的表面。为无滑膜肌腱滑膜化提供了实验形态学依据。     

无滑膜肌腱滑膜化的实验研究及其意义

无滑膜肌腱滑膜化的实验研究及其意义张正治，刘正津，糜建红第三军医大学重庆６３００３８本实验将家兔的鞘外无滑膜肌腱段放入膝关节腔内进行在体肌腱培养，另将胎儿无滑膜肌腱段体外与滑膜细胞联合培养，培养的肌腱段经组织学切片，免疫组化染色，扫描电镜观察。结果显...     

组织工程滑膜化肌健移植的实验研究

目的：观察用组织工程学方法形成的滑膜化肌鞘内移植后的形态变化。方法：在30只兔后肢的第2践腱鞘内移植入滑膜化肌腱段替代趾深屈肌腱。在不同时相点观察移植肌腱的愈合状况，粘连程度及屈趾功能。结果：滑膜化肌腱与受体肌腱之间形成了牢固的纤维性结合，粘连轻。结论：用组织工程学方法构建的滑膜化肌腱可以作为一种新的肌腱移植材料。     

Synoviolin基因修饰滑膜细胞预防肌腱粘连

背景:Synoviolin基因过表达,能使滑膜细胞过度生长和增殖,因此将Synoviolin基因转入膜细胞将有可能实现无滑膜肌腱滑膜化,从而有效防止肌腱粘连的发生。目的:观察Synoviolin基因修饰滑膜细胞预防肌腱粘连的影响。方法:将30只Leghom鸡随机分为实验组与对照组,建立肌腱损伤模型。实验组将含Synoviolin基因重组的腺病毒载体直接注入腱鞘残端表面。术后1,3,8周,行大体观察,组织学检查,生物力学测试。结果与结论:实验组第四趾肌腱缝合处粘连程度、第三趾肌腱的滑动距离及模拟主动屈曲度均优于对照组(P0.05);与对照组相比,实验组Ⅰ型胶原蛋白含量下降,Ⅲ型胶原蛋白比例下降;说明Synoviolin基因修饰滑膜细胞能使无滑膜肌腱滑膜化,提示利用Synoviolin基因修饰滑膜细胞是屈指肌腱术后预防肌腱粘连的有效方法之一。     

悬浮组织块法分离培养兔成纤维样滑膜细胞

目的:建立兔成纤维样滑膜细胞悬浮组织块分离培养法。方法:取正常兔膝关节滑膜组织,用悬浮组织块法分离培养兔成纤维样滑膜细胞,观察细胞生长状况及形态特征,取第3 代对数期细胞,CCK鄄8 法绘制其生长曲线,免疫荧光法检测波形蛋白的表达情况,并与组织块贴壁法进行比较。结果:两种方法体外培养得到的兔成纤维样滑膜细胞形态为长梭形纤维样,细胞生长力较旺盛,生长曲线为典型的“S冶形,免疫荧光检测显示第3 代细胞强表达波形蛋白。结论:悬浮组织块法可以分离培养出兔成纤样维滑膜细胞,方法简便,效率高,为体外分离培养兔成纤维样滑膜细胞提供了一种新的方法。     

不同来源人滑膜细胞体外培养的比较

目的对滑膜细胞分离培养的不同实验方法进行比较研究。方法取患者关节滑膜组织和游离的关节液,分别采用滑膜组织培养法和关节液培养法分离培养细胞,观察滑膜细胞的生长情况。结果采用组织培养法获得的滑膜细胞数量较多,细胞形态典型,增殖较快。关节液培养法培养的滑膜细胞．数量少,增殖慢,培养周期较长,所获得的滑膜细胞均能满足后续实验的需要。结论两种方法各有优缺点,可达到互为补充的效果。在临床上可根据病人的情况进行选择。     

兔膝骨关节炎成纤维样滑膜细胞分离培养方法的研究

目的 在传统细胞组织块培养方法的基础上进行改进,建立一种简单、可重复性操作的兔膝骨关节炎成纤维样滑膜细胞分离培养方法。方法 通过手术诱导建立兔膝骨关节炎模型,在无菌条件下,快速取出兔膝骨关节炎的滑膜组织,经过冲洗处理后剪成一定大小碎片,用无菌滤纸吸净组织上附着的水分,接种在培养皿后,进行观察、换液、传代、拍照。将原代培养的细胞传至第三代细胞接种于96孔板中,采用噻唑蓝（MTT）法分别在不同时间梯度检测各孔对应吸光度,制作生长曲线;采用免疫荧光染色法观察细胞中波形蛋白的表达情况。结果 培养的细胞为典型的梭形,符合成纤维样滑膜细胞的生长形态特征,免疫荧光染色显示波形蛋白强表达。结论 改进后的组织块培养方法简单、操作可重复性强,可以获得大量兔膝骨关节炎成纤维样滑膜细胞,为后续膝骨关节炎的机理研究提供了大量细胞支持。     

滑膜细胞中波形蛋白的形态学研究

用免疫组织化学、激光共聚焦扫描和粘附式细胞仪上进行三维图像重建，观察了波形蛋白在体外培养人腱鞘滑膜细胞内的分布。结果显示：波形蛋白呈海绵状立体结构，分布于整个细胞质空间；认为波形蛋白的分布与培养细胞在不同生长时期的形态变化密切相关。     

小肠黏膜下层与滑膜间充质干细胞的组织相容性

背景：小肠黏膜下层有抗微生物活性、良好的生物相容性及生物力学性能,能在体内快速降解,与半月板纤维软骨细胞的胞外基质相近。 目的：观察小肠黏膜下层与滑膜间充质干细胞是否有良好的组织相容性。 方法：先后采用物理方法与化学方法处理猪小肠黏膜下层,并进行苏木精-伊红染色和扫描电镜观察。制备小肠黏膜下层浸提液,行以下实验：①热源实验：在新西兰大白兔耳缘静脉注射小肠黏膜下层浸提液。②皮肤致敏实验：在新西兰大白兔背部皮内分别注射小肠黏膜下层浸提液、多聚甲醛溶液及生理盐水。③全身毒性实验：在新西兰大白兔耳缘静脉分别注射小肠黏膜下层浸提液与生理盐水。将小肠黏膜下层与成骨诱导的兔滑膜间充质干细胞共培养,以小肠黏膜下层单独培养为对照。 结果与结论：经物理处理过的小肠黏膜下层表面仍然存在小肠黏膜上皮细胞、脂肪细胞和一些其他细胞黏附;经化学方法处理后其残留的细胞数量明显减少,但主要结构和成分未被改变。小肠黏膜下层黏膜面较肌层面光滑,无细胞残留,表面纤维组织交错形成疏松的三维网状立体结构,孔隙率为80%。小肠黏膜下层是一种无毒、无刺激、无免疫原性,生物相容性很好的生物材料,与兔滑膜间充质干细胞具有良好的组织相容性。     

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...     

Synovial cell culture and tissue engineering of a tendon synovial cell biomembrane

Restrictive adhesions are a common complication of tendon injury and repair in the hand, resulting in severe dysfunction. Creating a barrier between the repair sites and surrounding tissue layers may prevent adhesions. We present the first stage in the process of developing a synovial biomembrane for this purpose. Synovial cells harvested from the Achilles tendon sheath and the knee joint of a Wistar albino rat were cultured for 2 weeks in culture medium, and then impregnated into a collagen type 1 matrix for another 2 weeks. Cells originating from both tendon and synovium demonstrated cell growth and layer formation on the surfaces of the matrix 2 weeks after impregnation. Alcian blue staining using Scott's method demonstrated the presence of acidic mucopolysaccharide, indicating hyaluronic acid (HA) production. This provides indirect evidence of functioning synovial cells on the membrane. It is possible to culture synovial cells and engineer a synoviocyte-collagen membrane that synthesizes endogenous HA. Application of this biomembrane to tendon repair sites may help to prevent adhesions after tendon repairs. Evaluation of this method on in vivo models is required.     

利用罗曼鸡肌腱滑液鞘体内构建趾深屈肌腱的生物力学及组织学研究

目的 探索肌腱滑液鞘对肌腱体内再生的作用。 方法 将36只罗曼鸡随机平均分为A、B两组。A组把左趾深屈肌腱滑液鞘的上、下、右侧分离,不切断趾深屈肌腱。同种异体脱细胞肌腱用部分分离的腱滑液鞘包裹,并固定在趾深屈肌腱左侧。B组直接把同种异体脱细胞肌腱固定在去除肌腱滑液鞘的趾深屈肌腱左侧。另取右趾深屈肌腱作为正常对照组。分别在第4、8、12周取材进行趾深屈肌腱最大载荷、弹性模量的生物力学检测及HE染色的组织学检测。结果 术后第4、8、12周时,除第4周A、B组在弹性模量方面差异无统计学意义,其余时间点A组最大载荷和弹性模量都大于B组（P<0.05）。随时间延长,A组胶原纤维增多,排列紧密,方向一致,炎性细胞浸润及纤维组织增生程度较轻。而B组胶原纤维数量逐渐减少、排列逐渐散乱,炎性细胞浸润、纤维组织增生程度明显高于A组。结论腱滑液鞘有助于肌腱再生,说明合适的体内环境对肌腱构建具有重要作用,本研究结果对临床上肌腱缺损病人找到合适肌腱替代物有积极作用。     

The accumulation of inflammatory cells in synovial sheath and epitenon during adhesion formation in healing rat flexor tendons.

The accumulation of inflammatory cells in synovial tissue was studied using indirect immunofluorescence assays on cell cultures and frozen tissue sections of healing rat digital flexor tendons. Flexor tendons were collected from rats 3, 7 and 14 days after crush injury. Tendon sheath and epithenon cells were isolated by sequential enzymic digestion and cultured for 2 days. Subpopulations of synovial and inflammatory cells were identified with MoAbs against cell surface glycoproteins present on B lymphocytes (CD45), T lymphocytes (CD2, CD4, CD8), macrophages (CD14) and endothelial cells. A phagocytosis assay was also used to identify macrophages. We report a substantial increase in the number of T lymphocytes (mainly helper/inducer) and phagocytotic cells with monocyte/macrophage surface markers in tendon sheath and epitenon 3 days after crush injury. The infiltration of inflammatory cells into synovial sheath and epitenon preceded an increase in fibronectin production by tendon cells which was seen 7 days after injury. To study the interaction between T lymphocytes and synovial cells in vitro, we established synovial fibroblast-like type B cell cultures and used stimulated and non-stimulated T lymphocytes in cell binding assays. We observed increased adhesiveness between unstimulated synovial cells and synovial cells previously cultured with activated and non-activated T lymphocytes. ELISA inhibition studies have shown an increase in fibronectin production by synovial fibroblasts co-cultured with stimulated CD4+ T lymphocytes. We suggest that the presence of inflammatory cells in synovial sheath and epitenon during tendon healing induces synovial fibroblasts and epitenon cells to increase their production of fibronectin, which provides a scaffold for subsequent adhesion formation.     

Ultrastructure of Human Tendon Sheath and Synovium: Implications for Tumor Histogenesis

Normal human tendon sheath and synovium were studied by scanning and transmission electron microscopy. The lining cells of the two tissues appear to be identical ultrastructurally. The most superficial cells (B-cells) possess long cytoplasmic extensions that clothe the membrane surface. Intermingled with deeper B-cells are the so-called A-cells, which have similar cytoplasmic features but lack long processes and instead have many filopodia. The frequent occurrence of intermediate forms indicates that the two cells form part of a morphologic spectrum. Comparison with cells of tumors that have been ascribed to synovium or tendon sheath (synovial sarcoma, epithelioid sarcoma, clear cell sarcoma) do not reveal any close similarities that might support a histogenetic relationship.     

A developmental study of the synovial membrane of the rat temporomandibular joint: changes in the three-dimensional configuration during postnatal development

The development of synovial membranes in the posterior synovial portion of the rat temporomandibular joint was studied and the three-dimensional structure of the posterior synovial portion reconstructed from sagittal semithin sections. Reconstructions showed that the synovial membrane expanded and that synovial folds increased in number and became complicated in shape with the growth of the joint. Using transmission-electron microscopy, it was observed that the synovial lining cells degenerated, that the synovial membrane split to make further synovial folds, and that the folded-end structures consisted of synovial lining cells that extended into the subsynovial connective tissue. It is suggested that in the development of the three-dimensional configuration of the synovial membrane, several processes proceed simultaneously to form the synovial folds: a splitting of the synovial membrane, infolding of the synovial membrane into the subsynovial connective tissue, and outgrowth of the synovial folds towards the synovial cavity.     

Decorin对僵直膝关节滑膜B型细胞增殖及I型胶原蛋白表达的影响

目的:探讨重组核心蛋白聚糖(decorin)对人僵直膝关节滑膜B型细胞的增殖和Ⅰ型前胶原蛋白(PcⅠ)mRNA表达及Ⅰ型胶原蛋白(ColⅠ)合成的影响,为decorin治疗人膝关节僵直提供理论依据。方法:分离培养人僵直膝关节滑膜B型细胞,分别加入不同浓度(0.1、5和10 mg/L)的decorin;培养24 h、48 h和72 h后用MTT法测定关节滑膜B型细胞的活力;用流式细胞术检测滑膜B型细胞的周期和凋亡;RT-PCR法检测滑膜B型细胞的PcⅠmRNA表达;Western blot检测ColⅠ的合成。结果:5和10 mg/L的decorin可有效降低滑膜B型细胞的活力(P0.05),明显增加处于G0/G1期细胞的百分比(P0.05),并下调PcⅠmRNA的表达,抑制ColⅠ的合成;同时实验组和对照组均未检测到终末期凋亡细胞。结论:Decorin可抑制人僵直膝关节滑膜B型细胞的增殖、PcⅠmRNA的表达和ColⅠ的合成,在防治膝关节僵直中可能起一定的作用。     

Overexpression of synoviolin facilitates the formation of a functional synovial biomembrane

Digital flexor tendon repair poses a significant challenge for hand surgeons. Currently, extrasynovial tendon grafts are frequently used in clinical settings to bridge flexor tendon defects. However, the healing process is always accompanied by postoperative adhesion. This is mostly due to the fact that no synovial membrane covers the extrasynovial tendon surface, in contrast to the intrasynovial tendon. In this study, we present an efficient method of developing a functional synovial biomembrane on the surface of the extrasynovial tendon. Synoviocytes were isolated from the knee joint of a Japanese white rabbit. After being infected with lentivirus, the over-expression of synoviolin in these synoviocytes was confirmed by semi-quantitative RT-PCR and western blotting. Cellular proliferation and increased hyaluronic acid secretion were confirmed in the synoviolin over-expressing synoviocytes by MTT-based method, cell cycle assays and ELISA. Furthermore, the synoviolin over-expressing synoviocytes were co-cultured with extrasynovial tendons that were harvested from the hind leg of rabbits. After being co-cultured in vitro for 3 and 7 days, these infected synoviocytes were found to accelerate the formation of a biomembrane on the tendon surface compared to the control group. More importantly, Alcian blue staining confirmed the ability of this cultured biomembrane to produce specific matrices containing acidic carboxyl mucopolysaccharides (mainly hyaluronic acid). All these results demonstrate that the over-expression of synoviolin stimulates the proliferation and HA secretion of synoviocytes and facilitates the formation of a functional synovial biomembrane.     

Blood supply of the flexor digital tendon in the hand and its clinical significance

Summary An anatomical study on the blood sources and vascularity of the flexor digital tendon was conducted in the upper extremities of fresh cadavers by means of arterial injection and meticulous dissection of the transparent tendon under the microscope. According to whether or not synovial membrane surrounded the tendon, the flexor digital tendon can be divided into 2 regions: non-synovial and synovial. The major intrinsic blood supply of the digital tendon was in the form of longitudinal vascular bundles, while the transverse anastomotic branches were short and sparse. The non-synovial region of the tendon was covered by paratenon and the vascular distribution of this region was uniform. In the synovial sheath, the blood vessels distributed only on the dorsal side, while the volar side was devoid of vessels. The profundus and superficialis tendons had an avascular zone at the proximal interphalangeal and metacarpophalangeal joints respectively. It was considered that the difference of the vascular architecture might be related to the mechanical force to which the tendon was subjected. The nutrition of tendon was discussed and the selection of tendon graft at operation was suggested.

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Vascularisation des tendons fléchisseurs des doigts au niveau de la main : signification clinique

Résumé Une étude anatomique des pédicules et de la vascularisation des tendons fléchisseurs des doigts a été réalisée sur cadavres frais, au moyen d'injections artérielles d'encre de chine, et d'une dissection méticuleuse sous microscope du tendon diaphanisé. Selon que la membrane synoviale entoure ou non le tendon, ce dernier peut être divisé en deux parties: partie non synoviale et partie synoviale. La vascularisation intrinsèque du tendon fléchisseur digital est essentiellement constituée par des éléments vasculaires longitudinaux, tandis que les anastomoses transversales sont courtes et éparses. La partie non synoviale du tendon est enveloppée par le paratendon et la distribution vasculaire dans cette région est uniforme; dans la gaine synoviale les vaisseaux sanguins se distribuent seulement du côté dorsal, tandis que le côté palmaire est dépourvu de vaisseaux. Les tendons fléchisseurs superficiels et profonds présentent une zone avasculaire au niveau des articulations interphalangienne et métacarpophalangienne. On peut penser que la différence d'architecture vasculaire peut être en relation avec les forces mécaniques auxquelles le tendon est soumis. Le mode de nutrition du tendon est discuté et le choix d'une greffe tendineuse en per-opératoire est suggéré.

     

Three-dimensional ultrastructure of synoviocytes in the knee joint of rabbits and morphological changes in osteoarthritis model

The synovial intima is composed of two types of synoviocytes: absorptive macrophages and secretory, fibroblast-like F cells. Many studies have tried to observe synoviocytes by scanning electron microscopy (SEM) but failed to reveal the entire shape of synoviocytes because they are deeply embedded in the interstitial matrix. The present study, primarily employing SEM observation of NaOH macerated samples, reveals the distribution and three-dimensional ultrastructure of the synoviocytes in the normal knee joint of rabbits, and the morphological changes of synoviocytes in an osteoarthritis model of this animal. F cells were broadly distributed throughout the synovial intima, while macrophages showed a restricted distribution on fatty tissues around the patella. F cells were classified into a flat type, which covered the surface of synovial membrane like an epithelium, and a dendritic type, which extended long processes to form a characteristic meshwork on the surface. The flat type predominated in regions adhering to the femur, while the dendritic type predominated in ambilateral parts of both the patella and tendon of the musculus quadriceps femoris, and on the peripatellar fatty tissue. Intermediate forms of flat and dendritic types appeared in middle regions between the patella and periphery of the joint capsule. In the synovial membrane of the osteoarthritis model, both types of synoviocytes increased in number and changed their morphology, indicating their elevated activities in absorption and secretion. It is suggested that the ultrastructural changes in synoviocytes reflect pathological conditions of the synovial membrane, and synoviocytes play important roles in the pathogenesis of osteoarthritis.