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实验性前盘移位对兔颅下颌关节硫酸化糖胺聚糖、透明质酸和连接蛋白影响的组织化学和免疫组织化学研究

Histochemical and immunohistochemical studies of the effects of experimental anterior disc displacement on sulfated glycosaminoglycans, hyaluronic acid, and link protein of the rabbit craniomandibular joint.

作者信息

Ali A M, Sharawy M

机构信息

School of Medicine, Medical College of Georgia, Augusta, USA.

出版信息

J Oral Maxillofac Surg. 1996 Aug;54(8):992-1003; discussion 1003-4. doi: 10.1016/s0278-2391(96)90399-7.

Abstract

PURPOSE

The purpose of this study was to determine the effects of surgically induced anterior disc displacement (ADD) on sulfated glycosaminoglycans (GAGs) such as keratan sulfate (KS), chondroitin-4-sulfate (C4S), and chondroitin-6-sulfate (C6S), hyaluronic acid (HA), and link protein (LP) of the rabbit craniomandibular joint (CMJ) using histochemical and immunohistochemical techniques.

MATERIALS AND METHODS

The right joint of 20 rabbits was exposed surgically, and all discal attachments were severed except for the posterior attachment. The disc was then repositioned anteriorly and sutured to the zygomatic arch. The left joint served as a sham-operated control. Ten additional joints were used as nonoperated controls. Deeply anesthetized rabbits were perfused with 2% buffered formalin 2 weeks (10 rabbits) or 6 weeks (10 rabbits) after surgery. Discs, bilaminar zones, condyles, and articular eminences were excised. Condyles and articular eminences were decalcified in ethylenediaminetetraacetic acid (EDTA). All tissues were sectioned at 10 microns in a cryostat. Sections were incubated with alcian blue and monoclonal antibodies directed against KS, C4S, C6S, HA, or LP. After incubation in the appropriate fluorescein isothiocyanate (FITC)-labeled secondary antibodies, tissue sections were studied under the fluorescence microscope.

RESULTS

The results showed a reduction in alcian blue staining and KS, C4S, C6S, HA, and LP immunostaining in the disc and articular surfaces at 2 weeks after induction of ADD. This reduction was followed by an increase in their immunostaining at 6 weeks. Also, there was a progressive increase in alcian blue staining, and KS, C4S, C6S, and HA immunostaining in the bilaminar zone at 2 and 6 weeks.

CONCLUSION

It was concluded that surgical induction of ADD in the rabbit CMJ leads to alterations in KS, C4S, C6S, HA, and LP content, consistent with similar changes accompanying osteoarthritis of other synovial joints.

摘要

目的

本研究旨在运用组织化学和免疫组织化学技术,确定手术诱导的前盘移位(ADD)对兔颅下颌关节(CMJ)中硫酸化糖胺聚糖(GAGs),如硫酸角质素(KS)、硫酸软骨素-4(C4S)、硫酸软骨素-6(C6S)、透明质酸(HA)和连接蛋白(LP)的影响。

材料与方法

对20只兔的右侧关节进行手术暴露,除后部附着外,切断所有盘附着。然后将盘向前重新定位并缝合至颧弓。左侧关节作为假手术对照。另外10个关节用作未手术对照。在术后2周(10只兔)或6周(10只兔),对深度麻醉的兔用2%缓冲福尔马林灌注。切除盘、双板区、髁突和关节结节。髁突和关节结节在乙二胺四乙酸(EDTA)中脱钙。所有组织在低温恒温器中切成10微米厚的切片。切片用阿尔辛蓝和针对KS、C4S、C6S、HA或LP的单克隆抗体孵育。在用适当的异硫氰酸荧光素(FITC)标记的二抗孵育后,在荧光显微镜下研究组织切片。

结果

结果显示,诱导ADD后2周,盘和关节表面的阿尔辛蓝染色以及KS、C4S、C6S、HA和LP免疫染色减少。这种减少之后,在6周时它们的免疫染色增加。此外,在2周和6周时,双板区的阿尔辛蓝染色以及KS、C4S、C6S和HA免疫染色逐渐增加。

结论

得出的结论是,兔CMJ中手术诱导的ADD导致KS、C4S-C6S、HA和LP含量发生改变,这与其他滑膜关节骨关节炎伴随的类似变化一致。

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