Immunohistochemical and Histochemical Analysis of Newly Formed Tissues in Root Canal Space Transplanted with Dental Pulp Stem Cells Plus Platelet-rich Plasma
Immunohistochemical and Histochemical Analysis of Newly Formed Tissues in Root Canal Space Transplanted with Dental Pulp Stem Cells Plus Platelet-rich Plasma
复制标题
牙髓干细胞加富含血小板血浆移植根管间隙中新形成组织的免疫组织化学和组织化学分析
DOI：
10.1016/j.joen.2014.05.010
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发表时间：
2014-10-01
影响因子：
4.2
通讯作者：
Zhang, Chengfei
中科院分区：
文献类型：
--
作者：
Zhu, Xiaofei;Wang, Yu;Zhang, Chengfei
前言：牙髓摘除术后根管内组织再生可通过移植自体牙髓干细胞和/或富血小板血浆来实现。然而，牙髓中新形成的组织的一致性；间隙仅通过组织学分析来检查。本研究旨在应用免疫组织化学和组织化学方法检测根管再生术后新生组织中的特异性标志物。方法：在我们先前的研究中，对4只成年犬的32个根管进行了牙髓再生，分别使用血块、牙髓干细胞移植、富含血小板的血浆或细胞与血浆的组合进行牙髓再生。检测牙周韧带组织中Periostin的表达，成牙本质细胞中巢蛋白和牙本质唾液蛋白的表达，骨组织中骨唾液蛋白和骨钙素的表达。样本还进行了抗酒石酸酸性磷酸酶(TRAP)染色，作为破骨细胞谱系的标志。结果：从牙周韧带到内管表面可见连续的Periostin阳性组织，在内管表面矿物质岛周围有弱的Periostin染色。TRAP、骨涎蛋白和骨钙素在椎管间隙也有阳性染色，提示有骨组织的存在。沿根管内表面的一层矿化组织呈TRAP阴性，提示该组织可能是牙骨质。在所有样本中，未观察到巢蛋白阳性反应，而牙本质唾液蛋白在牙周膜、牙本质小管和根管内纤维组织中检测到。4组间均无差异。结论：犬成熟根管再生术后形成的组织不是牙髓组织，而主要是牙周组织。
Introduction: Tissue regeneration in root canals after pulpectomy can be achieved by transplantation of autologous dental pulp stem cells and/or platelet-rich plasma. However, the identity of the newly formed tissue in the pulp; space has been only examined by histologic analysis. This study aimed to apply immunohistochemistry and histochemistry to detect specific markers in the newly generated tissues after root canal regenerative treatment. Methods: In our previous study, 32 root canals in 4 mature dogs were treated with a pulp regeneration procedure after pulpectomy using either blood clot, transplantation of dental pulp stem cells, platelet-rich plasma, or a combination of cells and plasma. In the present study, the tissues were examined for the expression of periostin to detect periodontal ligament tissue, nestin and dentin sialoprotein for odontoblasts, and bone sialoprotein and osteocalcin for bone tissues. Samples were also stained for tartrate-resistant acid phosphatase (TRAP) as a marker for osteoclastic lineages. Results: Continuous periostin-positive tissue was observed extending from the periodontal ligament into the inner canal surface in which the mineral islands were surrounded by weak periostin staining. There was also positive staining for TRAP, bone sialoprotein, and osteocalcin in the canal space, suggesting the presence of bone tissue. A layer of mineralized tissue along the inner surface of the root canal was negative for TRAP, suggesting the tissue likely to be cementum. In all samples, no nestin-positive reaction was observed, whereas dentin sialoprotein was detected in PDL, dentinal tubules, and intracanal fibrous tissues. There was no difference between any of the 4 groups. Conclusions: The tissues formed in the dog mature root canals after regenerative endodontic procedures are not pulp tissues but mainly periodontal tissues.