Dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHEDSCs), and periodontal ligament stem cells (PDLSCs) isolation, characterization and the effectiveness of allantoin as bioactive molecule for dental regeneration

IF 5.5 2区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE Journal of dentistry Pub Date : 2025-03-01 Epub Date: 2025-02-02 DOI:10.1016/j.jdent.2025.105604

Betül Mutlu Özçınar , Can Özükoç , Emrah Türkmen , Rabia Çakır

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Abstract

Introduction

Dental stem cells are valuable tools in regenerative medicine due to their pluripotency and self-renewal properties. This study aimed to investigate the effects of allantoin (Al) on Dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHEDSCs), and periodontal ligament stem cells (PDLSCs) regarding cytotoxicity, proliferation, wound healing, and osteogenic differentiation.

Methods

Human dental stem cells were isolated from three dental tissues using the explant culture method and cultured in DMEM-F12 medium supplemented with 15 % fetal bovine serum (FBS) and antibiotics. The cytotoxicity and proliferation of allantoin were assessed using the XTT cell viability assay at concentrations ranging from 0.25 to 5 mg/mL. Wound healing was evaluated through a scratch assay at 1 mg/mL, and osteogenic differentiation was assessed using Alizarin Red S staining at 0.5 mg/mL and 1 mg/mL.

Results

Al exhibited no cytotoxic effects across the tested concentrations. It enhanced cell proliferation, particularly in SHEDSCs at 5 mg/mL. DPSCs also showed significant improvement in wound healing in the scratch assay. At 1 mg/mL, Al inhibited osteogenic differentiation in DPSCs and PDLSCs, as indicated by reduced mineralization.

Conclusion

Al shows potential as a non-cytotoxic agent for enhancing the proliferation of dental stem cells, especially SHEDSCs. However, its limited effect on wound healing of SHEDSCs and PDLSCs and inhibition of osteogenic differentiation at higher concentrations suggest that further optimization is required for its application in bone regeneration.

Statement of Clinical Relevance

Evaluation of the effects of plant-based therapeutic compounds on various types of dental stem cells may have the potential to increase the success of stem cell-based therapies in clinical applications in regenerative dentistry.

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牙髓干细胞（DPSCs）、人脱落乳牙干细胞（SHEDSCs）和牙周韧带干细胞（PDLSCs）的分离、表征及尿囊素作为牙再生生物活性分子的有效性

牙干细胞因其多能性和自我更新的特性而成为再生医学的重要工具。本研究旨在探讨尿囊素（Al）对牙髓干细胞（DPSCs）、人脱落乳牙干细胞（SHEDSCs）和牙周韧带干细胞（PDLSCs）在细胞毒性、增殖、伤口愈合和成骨分化方面的影响。方法：采用外植体培养法从3种牙组织中分离出人牙干细胞，在添加15%胎牛血清（FBS）和抗生素的DMEM-F12培养基中培养。使用XTT细胞活力测定法评估尿囊素在0,25至5mg /mL浓度范围内的细胞毒性和增殖。在1 mg/mL浓度下通过划痕法评估伤口愈合情况，在0、5 mg/mL和1 mg/mL浓度下通过茜素红S染色评估成骨分化情况。结果：不同浓度的铝均无细胞毒作用。当浓度为5mg /mL时，它能促进细胞增殖，尤其是在SHEDSCs中。在划痕实验中，DPSCs也显示出明显的伤口愈合改善。在1mg /mL浓度下，Al抑制了DPSCs和PDLSCs的成骨分化，这表明矿化程度降低。结论：Al作为一种非细胞毒性药物具有促进牙干细胞，尤其是SHEDSCs增殖的潜力。然而，其对SHEDSCs和PDLSCs伤口愈合的作用有限，高浓度下对成骨分化的抑制作用也有限，因此其在骨再生中的应用还需要进一步优化。临床相关性声明：评估植物性治疗化合物对不同类型牙科干细胞的作用可能有潜力增加干细胞治疗在再生牙科临床应用中的成功。

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来源期刊

Journal of dentistry 医学-牙科与口腔外科

CiteScore

7.30

自引率

11.40%

发文量

349

审稿时长

35 days

期刊介绍： The Journal of Dentistry has an open access mirror journal The Journal of Dentistry: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal of Dentistry is the leading international dental journal within the field of Restorative Dentistry. Placing an emphasis on publishing novel and high-quality research papers, the Journal aims to influence the practice of dentistry at clinician, research, industry and policy-maker level on an international basis. Topics covered include the management of dental disease, periodontology, endodontology, operative dentistry, fixed and removable prosthodontics, dental biomaterials science, long-term clinical trials including epidemiology and oral health, technology transfer of new scientific instrumentation or procedures, as well as clinically relevant oral biology and translational research. The Journal of Dentistry will publish original scientific research papers including short communications. It is also interested in publishing review articles and leaders in themed areas which will be linked to new scientific research. Conference proceedings are also welcome and expressions of interest should be communicated to the Editor.