Treatment of gingival defects with gingival mesenchymal stem cells derived from human fetal gingival tissue in a rat model

Jing Li,Bao-Hua Xu,Hong-Lin Liu,Hong Li,Li-Hong Ge,Wen-Jian Zhang,Zhen Xu,Kai Zhang,Shi-Qing Xu,Jin-Ning Lou

doi:10.1186/s13287-017-0751-7

Jing Li, Bao-Hua Xu + Show 8 more

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https://doi.org/10.1186/s13287-017-0751-7

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Abstract

BackgroundThe study aimed to evaluate the efficacy and safety of gingival mesenchymal stem cells (GMSCs) from human fetal gingival tissue used for treating gingival defects in a rat model.MethodsGMSCs were isolated from human fetal gingival tissue and identified by flow cytometry for nestin, Oct4, vimentin, NANOG, CD105, and CD90. The immunogenicity of GMSCs was analyzed by mixed lymphocyte reactions; the tumorigenicity of GMSCs was evaluated by xenotransplanting into nude mice. The gingival defect animal model was established by mechanical resection in rats. GMSCs were transplanted into the defective area, and the regeneration of gingival tissue was observed twice weekly. Four weeks after transplantation, the gingival tissue was surgically cut down, and the graft was analyzed by immunohistochemistry staining for human mitochondrial antigens and rat CD3 and CD20.ResultsGMSCs from human fetal gingival tissue positively expressed nestin, Oct4, vimentin, NANOG, CD105, and CD90. There was no cell aggregation after mixed lymphocyte reactions, and interleukin-2 did not increase. Inoculation of GMSCs into nude mice for 6 months showed no tumor formation. GMSCs were transplanted into the gingiva defects of rats. One week after transplantation, the defect area was reduced, and after 3 weeks the morphology and color of local gingival tissue was similar to normal gingival tissue, and gingival height was the same as the normal control group.ConclusionsUsing GMSCs from human fetal gingival tissue to treat gingival defects is a safe and effective innovative treatment method.

Highlights

The study aimed to evaluate the efficacy and safety of gingival mesenchymal stem cells (GMSCs) from human fetal gingival tissue used for treating gingival defects in a rat model
Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM)/F12 containing 10% fetal calf serum for stem cells (Biochrom, Germany), 40 μg/L leukemia inhibitor factor (LIF; PeproTech, USA), 10 μg/L basic fibroblast growth factor, and 10 μg/L epidermal growth factor (EGF; PeproTech) in 25-cm2 T-flasks at 37 °C in 5% CO2
Culture and identification of human fetal GMSCs Human fetal GMSCs adhered to the bottom of flasks after overnight culture

Summary

Introduction

The study aimed to evaluate the efficacy and safety of gingival mesenchymal stem cells (GMSCs) from human fetal gingival tissue used for treating gingival defects in a rat model. Periodontitis is an inflammatory and destructive disease that occurs in the gingiva, periodontal ligaments, alveolar bone, and cementum, and is one of the oldest and most common oral diseases. The incidence of periodontal disease in different countries varies from 50 to 90% [1, 2] and is the main cause of tooth loss in adults. The typical feature of the stationary phase of periodontitis is defects of the gingiva and other periodontal tissues. The clinical symptoms caused by root exposure often require surgical methods, such as the laterally positioned flap, the free gingival graft, and guide tissue regeneration, to cover the exposed root surface and improve clinical symptoms. Surgical treatment often results in trauma, covers a limited treatment area, requires adequate

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