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再生性和传统牙周手术后猴与人牙周组织中细胞角蛋白的表达模式

Patterns of cytokeratin expression in monkey and human periodontium following regenerative and conventional periodontal surgery.

作者信息

Sculean A, Berakdar M, Pahl S, Windisch P, Brecx M, Reich E, Donos N

机构信息

Department of Periodontology and Conservative Dentistry, University of Saarland, Homburg, Germany.

出版信息

J Periodontal Res. 2001 Aug;36(4):260-8. doi: 10.1034/j.1600-0765.2001.036004260.x.

DOI:10.1034/j.1600-0765.2001.036004260.x
PMID:11519700
Abstract

The pattern of cytokeratin expression has been extensively described in the normal and inflamed periodontium. However, there is no information regarding the pattern of cytokeratin expression in the periodontium which has been reformed following regenerative periodontal surgery. The aim of the present investigation was to evaluate the pattern of cytokeratin expression in the reformed human and monkey periodontium following regenerative and conventional periodontal surgery. In 3 monkeys, acute fenestration-type and chronic intrabony defects were treated with guided tissue regeneration (GTR), enamel matrix proteins (EMD), or coronally repositioned flap surgery (control). After a healing period of 5 months, the animals were sacrificed and perfused with 10% buffered formalin for fixation. Specimens containing the defects and surrounding tissues were dissected free, decalcified in EDTA and embedded in paraffin. Histological sections were cut with the microtome set at 3 microm. The sections were alternatively stained either with hematoxylin and eosin, or immunohistochemically by using one of the broad range monoclonal antibodies 34betaE 12 (for cytokeratins 1, 5, 10 and 14) or KL 1 (for cytokeratins 1, 2, 5, 6, 7, 8, 10, 11, 16 and 19), or one of the individual monoclonal antibodies LL025 (for cytokeratin 16), DC 10 (for cytokeratin 18), A53-B/A2 (for cytokeratin 19). Twelve patients, each displaying one deep intrabony defect scheduled for extraction due to advanced periodontitis or prosthetic reasons, were treated as described above. Following a healing period of 6 months, the teeth were extracted together with some of their surrounding soft and hard tissues. The histological and immunohistochemical processing of the human biopsies was identical to that described in monkeys. The results revealed that both the normal non-treated (original) monkey and human junctional epithelium stained strongly with all of the monoclonal antibodies used. The reformed junctional epithelium displayed the same cytokeratin expression pattern as the non-treated junctional epithelium. No differences regarding the cytokeratin expression pattern of the junctional epithelium were found between the treatments and types of healing (i.e. regenerative, through a new periodontal ligament, or reparative through a long junctional epithelium). In the intact periodontal ligament, the epithelial rests of Malassez displayed a comparable cytokeratin expression pattern to that of the junctional epithelium. However, no expression of cytokeratins was seen in the newly formed periodontal ligament. In such specimens, cytokeratin expression was observed only until the borderline between the regenerated cementum and the epithelial downgrowth. It was concluded that: a) the reformed junctional epithelium, following any type of surgical procedure, displays a similar pattern of cytokeratin expression to the original junctional epithelium; b) in the newly formed periodontal ligament, no expression of cytokeratins is present; and c) the epithelial rests of Malassez do not seem to reform after regenerative periodontal surgery.

摘要

细胞角蛋白在正常和炎症性牙周组织中的表达模式已被广泛描述。然而,关于再生性牙周手术后改建的牙周组织中细胞角蛋白的表达模式尚无相关信息。本研究的目的是评估再生性和传统牙周手术后人类和猴改建牙周组织中细胞角蛋白的表达模式。对3只猴的急性开窗型和慢性骨内缺损分别采用引导组织再生术(GTR)、釉基质蛋白(EMD)或冠向复位瓣手术(对照)进行治疗。经过5个月的愈合期后,处死动物,并用10%缓冲甲醛灌注固定。将包含缺损及周围组织的标本分离出来,在乙二胺四乙酸(EDTA)中脱钙,然后石蜡包埋。用切片厚度设置为3微米的切片机制作组织学切片。切片交替进行苏木精和伊红染色,或使用广谱单克隆抗体34βE12(针对细胞角蛋白1、5、10和14)或KL1(针对细胞角蛋白1、2、5、6、7、8、10、11、16和19),或个别单克隆抗体LL025(针对细胞角蛋白16)、DC10(针对细胞角蛋白18)、A53 - B/A2(针对细胞角蛋白19)之一进行免疫组织化学染色。12例患者,每人有一个因重度牙周炎或修复原因计划拔除的深骨内缺损,按上述方法进行治疗。经过6个月的愈合期后,将牙齿连同其周围的一些软硬组织一并拔除。人类活检组织的组织学和免疫组织化学处理与猴的处理相同。结果显示,正常未处理(原始)的猴和人类结合上皮对所有使用的单克隆抗体均呈强阳性染色。改建的结合上皮显示出与未处理的结合上皮相同的细胞角蛋白表达模式。在治疗方法和愈合类型(即通过新的牙周膜再生或通过长结合上皮修复)之间,未发现结合上皮细胞角蛋白表达模式存在差异。在完整的牙周膜中,马拉瑟上皮剩余显示出与结合上皮相当的细胞角蛋白表达模式。然而,在新形成的牙周膜中未观察到细胞角蛋白的表达。在这类标本中,仅在再生牙骨质与上皮下延的边界处观察到细胞角蛋白表达。得出以下结论:a)任何类型手术操作后改建的结合上皮,其细胞角蛋白表达模式与原始结合上皮相似;b)在新形成的牙周膜中,不存在细胞角蛋白表达;c)马拉瑟上皮剩余在再生性牙周手术后似乎不会改建。

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