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生物陶瓷对人牙髓干细胞牙本质/成骨分化的影响。

Effect of Biodentine on Odonto/Osteogenic Differentiation of Human Dental Pulp Stem Cells.

作者信息

Wang Xuerong, Cai Yixin, Zhang Min, Xu Junchen, Zhang Chengfei, Li Jin

机构信息

Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, Jiangsu Province Key Laboratory of Oral Diseases, Department of Geriatric Dentistry, The Affiliated Stomatological Hospital of Nanjing Medical University, 136 Hanzhong Road, Nanjing 210029, China.

Department of Endodontics, Yancheng Stomatological Hospital, Yancheng 224000, China.

出版信息

Bioengineering (Basel). 2022 Dec 21;10(1):12. doi: 10.3390/bioengineering10010012.

Abstract

This study aims to compare the biological characteristics of human dental pulp stem cells (hDPSCs) isolated from different-aged populations and examine the effects of Biodentine on proliferation and odonto/osteogenic differentiation of hDPSCs isolated from the elderly in vitro. hDPSCs were isolated from three different-aged populations: group A (≤18 years old), group B (19−59 years old), and group C (≥60 years old). The adhesion, proliferation, odonto/osteogenesis, and senescence were compared. The optimal concentration of aqueous Biodentine extract was determined by CCK-8 assay, alkaline phosphatase (ALP), and alizarin red staining (ARS). The effect of Biodentine on odonto/osteogenic gene and protein expression of hDPSCs in each group was evaluated by quantitative real-time PCR (QRT-PCR) and Western blot. hDPSCs were successfully isolated from three different-aged populations. Flow cytometry revealed that all isolated hDPSCs were positive for CD73 (>90%), CD90 (>90%), CD146 (<30%), and negative for CD45 (<1%). There existed an age-related decline in proliferation, odonto/osteogenic gene expression, and S-phase fraction (p < 0.05), an increase in senescence genes and p21 and p16 expression, and time needed for cell adhesion. Biodentine promoted hDPSC proliferation and mineralization in each group, particularly at a concentration of 0.2 mg/mL. Biodentine markedly enhanced odonto/osteogenesis-related gene and protein expression in each group (p < 0.05). hDPSCs can be obtained from populations of all ages. Though there is an age-related decline in their biological properties, hDPSCs from the elderly still maintain certain proliferation and multidirectional differentiation abilities. Biodentine can significantly promote the proliferation and odonto/osteogenic differentiation of hDPSCs isolated from the elderly over 60 years old, which could be considered a pulp capping material for vital pulp therapy in the elderly. Nevertheless, the efficacy of Biodentine in clinical application has to be further studied.

摘要

本研究旨在比较从不同年龄人群分离的人牙髓干细胞(hDPSCs)的生物学特性,并检测Biodentine对体外分离的老年hDPSCs增殖及成牙本质/成骨分化的影响。hDPSCs从三个不同年龄人群中分离:A组(≤18岁)、B组(19 - 59岁)和C组(≥60岁)。比较了细胞的黏附、增殖、成牙本质/成骨能力及衰老情况。通过CCK - 8法、碱性磷酸酶(ALP)及茜素红染色(ARS)确定Biodentine水提取物的最佳浓度。采用定量实时PCR(QRT - PCR)和蛋白质免疫印迹法评估Biodentine对各组hDPSCs成牙本质/成骨相关基因和蛋白表达的影响。成功从三个不同年龄人群中分离出hDPSCs。流式细胞术显示,所有分离的hDPSCs CD73阳性(>90%)、CD90阳性(>90%)、CD146阴性(<30%)、CD45阴性(<1%)。增殖、成牙本质/成骨相关基因表达及S期比例存在与年龄相关的下降(p < 0.05),衰老相关基因及p21和p16表达增加,细胞黏附所需时间增加。Biodentine促进各组hDPSC增殖和矿化,尤其在浓度为0.2 mg/mL时。Biodentine显著增强各组成牙本质/成骨相关基因和蛋白表达(p < 0.05)。hDPSCs可从各年龄段人群中获得。虽然其生物学特性存在与年龄相关的下降,但老年hDPSCs仍保持一定的增殖和多向分化能力。Biodentine可显著促进60岁以上老年人分离的hDPSCs增殖及成牙本质/成骨分化,可考虑作为老年人活髓治疗的盖髓材料。然而,Biodentine在临床应用中的疗效仍需进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3591/9854827/08e46a8db465/bioengineering-10-00012-g001.jpg

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