人牙髓淋巴管相关标志物的表达和超微结构分析

上海口腔医学 ›› 2013, Vol. 22 ›› Issue (6): 618-622.

人牙髓淋巴管相关标志物的表达和超微结构分析

兰卫东¹, 董迎春², 刘锐³

(1.南京大学医学院附属口腔医院牙体牙髓病科，2.麻醉科, 3.口腔颌面外科，江苏南京 210008)

收稿日期:2013-07-02 修回日期:2013-09-05 出版日期:2013-04-12 发布日期:2013-04-12
通讯作者: 董迎春，Tel: 025-83620337，E-mail:dongyingchun1001@hotmail.com
作者简介:兰卫东(1970-)，男，硕士，副主任医师，E-mail:lwd700123@sina.com
基金资助:
国家自然科学基金(81100768)；南京市医学科技发展专项资金资助项目(YKK11040,QRX11123 )

Specific markers and ultrastructure of lymphatic vessel in healthy human dental pulp

LAN Wei-dong¹, DONG Ying-chun², LIU Rui³

1.Department of Endodontics, 2. Department of Anesthesiology, 3.Department of Oral and Maxillofacial Surgery, Institute and Hospital of Stomatology, Nanjing University Medical School. Nanjing 210008, Jiangsu Province, China

Received:2013-07-02 Revised:2013-09-05 Online:2013-04-12 Published:2013-04-12
Supported by:
Supported by National Natural Science Foundation of China (81100768) and Key Project

摘要/Abstract

摘要： 目的：探讨健康牙髓中是否存在淋巴管。方法：选择因正畸拔除的牙30颗，取出牙髓，以健康牙龈组织做阳性对照，利用淋巴管特异性标志物D2-40和LYVE-1以及血管标志物CD31和CD34进行免疫组织化学染色，Western印迹法检测D2-40的表达，透射电镜进行超微结构分析。结果：健康牙髓组织中，管腔结构的内皮细胞均显示CD31和(或)CD34阳性，D2-40除成牙本质细胞弱阳性表达外，管腔结构均阴性表达。Western印迹法检测显示，用胶原酶去除牙髓成牙本质细胞后，D2-40所对应的分子量40 kDa的条带消失。牙髓管腔结构的内皮细胞电镜检测显示出血管而不是淋巴管的特征，即由单层内皮细胞组成，外有周细胞围绕，基底膜完整。结论：在人正常牙髓组织中可能不存在淋巴管，淋巴系统是否参与牙髓炎症的引流过程尚有待进一步研究。

关键词: 牙髓, 血管, 淋巴管, D2-40, 超微结构

Abstract: PURPOSE: To explore the existence of lymphatic vessels in healthy human dental pulp. METHODS: Thirty healthy human dental pulps were obtained from non-carious premolars removed for orthodontic reasons. Immunohistochemistry was performed using the antibodies specific for lymphatic endothelium such as D2-40 and LYVE-1, and for vascular endothelial cell such as CD31 and CD34. The expression of D2-40 was detected by Western blotting and ultrastructure was examined by transmission electron microscopy. RESULTS: In healthy human dental pulps, we failed to detect any reactivity for the lymphatic markers D2-40 and LYVE-1 in the observed vessels. These vessels were positive stained by blood endothelial markers CD34 and CD31. Odontoblasts were weakly stained with D2-40. Western blotting performed on collagenase-treated human dental pulps did not show a band at 40 kDa, corresponding to the molecular weight of the lymphatic marker D2-40. Transmission electron microscopy indicated that vessels in dental pulp consisted of endothelial monolayer surrounded by pericytes and complete basement membrane, which were typical ultrastructural characteristics of blood vessels rather than lymphatic vessel. CONCLUSIONS: Human dental pulp does not contain true lymphatic vessels under healthy conditions. Whether lymphatic system is involved in dental pulp interstitial fluid circulation during inflammation deserved further study.

Key words: Dental pulp, Blood vessel, Lymphatic vessel, D2-40, Ultrastructure

中图分类号:

R780.2

兰卫东, 董迎春, 刘锐. 人牙髓淋巴管相关标志物的表达和超微结构分析[J]. 上海口腔医学, 2013, 22(6): 618-622.

LAN Wei-dong, DONG Ying-chun, LIU Rui. Specific markers and ultrastructure of lymphatic vessel in healthy human dental pulp[J]. Shanghai Journal of Stomatology, 2013, 22(6): 618-622.

参考文献

[1] Matsumoto Y, Kato S, Miura M, et al. Fine structure and distribution of lymphatic vessels in the human dental pulp: a study using an enzyme-histochemical method [J]. Cell Tissue Res, 1997, 288(1): 79-85.
[2] Sawa Y, Yoshida S, Ashikaga Y, et al. Immunohistochemical demonstration of lymphatic vessels in human dental pulp [J]. Tissue Cell, 1998, 30(5): 510-516.
[3] Matsumoto Y, Zhang B, Kato S. Lymphatic networks in the periodontal tissue and dental pulp as revealed by histochemical study [J]. Microsc Res Tech, 2002, 56(1): 50-59.
[4] Marchetti C, Poggi P. Lymphatic vessels in the oral cavity: different structures for the same function [J]. Microsc Res Tech, 2002, 56(1): 42-49.
[5] Oehmke MJ, Knolle E, Oehmke HJ. Lymph drainage in the human dental pulp [J]. Microsc Res Tech, 2003, 62(3): 187-191.
[6] Pimenta FJ, SAR, Gomez RS. Lymphangiogenesis in human dental pulp [J]. Int Endod J, 2003, 36(12): 853-856.
[7] Gerli R, Secciani I, Sozio F, et al. Absence of lymphatic vessels in human dental pulp: a morphological study[J]. Eur J Oral Sci, 2010, 118(2): 110-117.
[8] Virtej A, L?es S, Iden O, et al. Vascular endothelial growth factors signalling in normal human dental pulp: a study of gene and protein expression [J]. Eur J Oral Sci, 2013, 121(2): 92-100.
[9] 宫艳红, 汪竹平, 王铎, 等. 改良5’-核苷酸酶-碱性磷酸酶双染色法观察成牙本质细胞层毛细淋巴管 [J]. 牙体牙髓牙周病学杂志, 2010, 20(8): 430-436.
[10] Berggreen E, Haug SR, Mkonyi LE, et al. Characterization of the dental lymphatic system and identification of cells immunopositive to specific lymphatic markers [J]. Eur J Oral Sci, 2009, 117(1): 34-42.
[11] Niakosari F, Kahn HJ, Marks A, et al. Detection of lymphatic invasion in primary melanoma with monoclonal antibody d2-40: a new selective immunohistochemical marker of lymphatic endothelium [J]. Arch Dermatol, 2005, 141(4): 440-444.
[12] Ozmen F, Ozmen MM, Ozdemir E, et al. Relationship between lyve-1, vegfr-3 and cd44 gene expressions and lymphatic metastasis in gastric cancer[J]. World J Gastroenterol,2011,17(27): 3220-3228.
[13] Jennings RN, Miller MA, Ramos-Vara JA. Comparison of cd34, cd31, and factor Ⅷ-related antigen immunohistochemical expression in feline vascular neoplasms and cd34 expression in feline nonvascular neoplasms[J].Vet Pathol,2012,49(3): 532-537.
[14] Martin A, Gasse H, Staszyk C. Absence of lymphatic vessels in the dog dental pulp: an immunohistochemical study [J]. J Anat, 2010, 217(5): 609-615.
[15] Takahashi S, Ambe K, Kon H, et al. Immunohistochemical investigation of lymphatic vessel formation control in mouse tooth development: lymphatic vessel-forming factors and receptors in tooth development in mice [J]. Tissue Cell, 2012, 44(3): 170-181.
[16] 张武, 汪竹平, 王铎, 等. 急性炎症牙髓中毛细淋巴管的超微结构 [J]. 上海口腔医学, 2000, 9(3): 158-160.
[17] 张武, 汪竹平, 王铎, 等. 慢性牙髓炎毛细淋巴管的透射电镜观察 [J]. 实用口腔医学杂志, 2005, 21(5): 702-704.