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Histlogical Examination of Periodontal Regeneration by Implantation of Cultured Periodontal Ligament Cell

培养牙周膜细胞植入牙周再生的组织学检查

基本信息

批准号：
09671942
负责人：
SUGAYA Tsutomu
金额：
$ 1.41万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

In the previous study, it was suggested that the cultured and implanted periodontal ligament cell had the regenerative potential for the periodontal ligament and cementum. But the kinetic aspects and the function of the cultured and implanted cell were not cleared. The purpose of this study is to evaluate the kinetic aspects of cultured and implanted periodontal ligament cells. In the experimental group, periodontal ligament cells were cultured and labeled on the dentin blocks in Minimum Essential Medium (MEM), supplemented with bromodeoxyuridine (BrdU) and implanted on the syngeneic rats calvaria defects. In the control group, dentin blocks without cultured cells were implanted. The samples were prepared for histological examination on 7, 14, 21 and 28 daysafter implantation. Specimens were made with Historesin plus, immunostained with anti-BrdU monoclonal antibody, and stained for alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP). In the control group, on 7 day TRAP-positive cells were detected, and TRAP-positive and ALP -positive cells were observed on 14 day. Ankylosis was also observed on 14 day, which was advanced on 21 and 28 day. In the experimental group, results were similar to those of the control group on the 7 day. But on 14, 21and 28 day, cellular cementum-like tissue was observed on the dentin blocks in some of the specimens. BrdU positive fibroblast-like cells, ALP-positive cells and TRAP-positive cells were also detected around the newly formed cellular cementum-like tissue. These results suggest that the cultured and implanted periodontal ligament cells are able to persist up to 28 days after implantation, and may be concerned with the formation of new cementum-like tissue and inhibit ankylosis.

以往的研究认为，牙周膜细胞培养后植入牙周膜组织中，具有再生牙周膜和牙骨质的潜能。但对体外培养和植入的细胞的动力学和功能尚不清楚。本研究的目的是评估牙周膜细胞培养和植入的动力学方面。实验组将牙周膜细胞在添加溴脱氧尿苷（BrdU）的最低必需培养基（MEM）中培养并标记于牙本质块上，种植于同系大鼠颅骨缺损处。对照组植入不含培养细胞的牙本质块。分别于植入后7、14、21、28天取材进行组织学检查。用Historesin plus制作标本，用抗BrdU单克隆抗体进行免疫染色，并进行碱性磷酸酶（ALP）和抗酒石酸酸性磷酸酶（TRAP）染色。在对照组中，在第7天检测到TRAP阳性细胞，并且在第14天观察到TRAP阳性细胞和ALP阳性细胞。第14天也观察到关节强直，21和28天进展。实验组在第7天的结果与对照组相似。14、21、28天，部分牙本质块表面出现细胞状牙骨质样组织。在新生的细胞性牙骨质样组织周围可见BrdU阳性的成纤维细胞样细胞、ALP阳性细胞和TRAP阳性细胞。这些结果表明，培养和植入的牙周膜细胞能够在植入后持续长达28天，并可能与新的牙骨质样组织的形成和抑制强直有关。