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Formation of Human Periodontal Ligament Cell Spheroids on Chitosan Films
Published on: June 19, 2019
In vitro proliferation of periodontal ligament-like tissue on extracted teeth
1Division of Orthodontics, Department of Oral Function & Restoration, Graduate School of Dentistry, Kanagawa Dental University, Kanagawa, Japan.
Objective:
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.
Design:
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.
Result:
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.
Conclusions:
We have developed a useful culture method to stimulate the proliferation of cells in PDL-like tissue on the roots of extracted teeth.
