Iwata T, Mino C, Kawata T
Division of Orthodontics, Department of Oral Function & Restoration, Graduate School of Dentistry, Kanagawa Dental University, Kanagawa, Japan.
Division of Orthodontics, Department of Oral Function & Restoration, Graduate School of Dentistry, Kanagawa Dental University, Kanagawa, Japan.
Arch Oral Biol. 2017 Mar;75:31-36. doi: 10.1016/j.archoralbio.2016.11.015. Epub 2016 Nov 30.
Transplantation of autologous teeth is a routine component of orthodontic treatment. The aim of this study was to develop a method for the regeneration of damaged periodontal ligament (PDL) on extracted teeth using a three-dimensional culture system.
We used the maxillary first premolars or third molars extracted from patients for orthodontic treatment. The extracted teeth were stained with toluidine blue to measure the residual PDL area. After confirming damage of the periodontal tissue on the root surface of the extracted teeth, we tried to regenerate the periodontal tissue. Other extracted teeth were inserted into a cell strainer filled with cellulose-based carrier materials to regenerate the periodontal tissue. The strainer was then placed in a 90-mm culture dish filled with culture medium and incubated at 37°C and 5% CO2 for about 1 month. The cultured teeth were observed under a stereomicroscope and examined by scanning electron microscopy (SEM), and were stained to detect alkaline phosphatase (ALP) activity.
Toluidine blue staining revealed that the residual periodontal membrane covered an average of 50.4% of the root surface area of each tooth. After culturing extracted teeth with our culture system, globular structures were found on the entire tooth root surface by stereomicroscopy, and PDL-like filamentous tissue was also detected by SEM. The entire tooth root surfaces of the cultured teeth were positive for ALP activity.
We have developed a useful culture method to stimulate the proliferation of cells in PDL-like tissue on the roots of extracted teeth.
自体牙移植是正畸治疗的常规组成部分。本研究的目的是开发一种利用三维培养系统使拔除牙上受损的牙周膜(PDL)再生的方法。
我们使用从正畸治疗患者口中拔除的上颌第一前磨牙或第三磨牙。将拔除的牙齿用甲苯胺蓝染色以测量残余牙周膜面积。在确认拔除牙牙根表面的牙周组织受损后,我们尝试使牙周组织再生。将其他拔除牙插入装有纤维素基载体材料的细胞筛中以再生牙周组织。然后将细胞筛置于装有培养基的90毫米培养皿中,在37°C和5%二氧化碳条件下孵育约1个月。在体视显微镜下观察培养的牙齿,并用扫描电子显微镜(SEM)进行检查,同时进行染色以检测碱性磷酸酶(ALP)活性。
甲苯胺蓝染色显示,残余牙周膜平均覆盖每颗牙齿牙根表面积的50.4%。用我们的培养系统培养拔除牙后,通过体视显微镜在整个牙根表面发现了球状结构,并且通过SEM也检测到了类似PDL的丝状组织。培养牙齿的整个牙根表面ALP活性呈阳性。
我们开发了一种有用的培养方法,可刺激拔除牙牙根上类似PDL组织中细胞的增殖。