Proteoglycans in gingival crevicular fluid as a maker of bone destruction

研究表明硫酸软骨素蛋白多糖（CS-PG）与鹿牙周组织的代谢和破坏有关。本研究是为了证实CS-PG作为骨破坏的制造者的假设。1. 提取GCF、牙龈和血清蛋白，SDS-PAGE分离，Western blotting法转移到硝化纤维素上。用单克隆抗体对与pg相关的C4S和C6S表位进行免疫染色，GcF中两种pg的主要条带在电泳迁移率上与这些条带相对应，而GcF中少数微弱染色条带与龈相对应。2. 在醋酸纤维素膜上用双向电泳法分离龈下牙石中的硫酸酸化pg。结果表明，CS-PG是脱钙球茎中唯一的硫酸化PG。此外，所有CS-PG均为C4S-PG， HPLC法未检出C6S异构体。3. ELISA法检测GCF中与骨胶原特异性交联的吡啶啉（Pyr）和脱氧吡啶啉（DPyr），这些交联在实验性牙周炎急性期增加，在慢性期减少。这一发现与以往炎症急性期GCF中C4S-PG含量升高的结果一致。

It has been demonstrated that chondoroitin sulfate proteoglycan(CS-PG) in assosiated with th emetabolism and destruction of deerer periodontal tissue. This study was porformed to confirm the hypothesis of CS-PG as a maker of bone destruction. 1. the extracted proteins from GCF, gingiva and serum were separated by SDS-PAGE, transfered to nitrocellulose by the method of Western blotting. Immunostaining was performed using monoclonal antivodies which recognize epitopes in C4S and C6S assosiated with PG.The major bands of both PGs in GcF corresponded in electrophoretic mobikity to that of these rum, while a few faintly stained bands in GCF Correspond to that of the gingica. 2. The sulfated PGs from human subgingival calculus were separated on Cellulise acetate membrane using two dimensional electrophoresis. The results showed that CS-PG is the sole sulfated PG in decalcified Clculus. In addition, all of CS-PG was C4S-PG, not detectable C6S isomer in sample by HPLC procedure. 3. Pyridinoline(Pyr) and Deoxypyridinoline(DPyr), spesific crosslinks to bone collagen, were detected in GCF by ELISA technuque, These crosslinks were found to increase in the acute phase and decrease in the chronic phase during experimental periodontitis. Such a findings is accord with the previous results in which the content of C4S-PG in GCF increase in acute phase of inflammation.