Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand.
School of Pharmacy, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand.
Int J Mol Sci. 2022 Aug 20;23(16):9425. doi: 10.3390/ijms23169425.
This investigation aimed to isolate and culture human dental pulp cells from carious teeth (cHDPCs) and compare their growth characteristics, colony-forming efficiency, mineralization potential and gene expression of Toll-like receptors (TLR)-2, TLR-4, TLR-9, tumour necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-8, IL-17A, 1L-17R, IL-23A, nuclear factor-kappa B (NF-κB), mitogen-activated protein kinase (MAPK1), dentin matrix protein (DMP)-1, dentin sialophospho protein (DSPP), sex determining region Y-box 2 (SOX2) and marker of proliferation Ki-67 (MKi67) with cells isolated from healthy or non-carious teeth (ncHDPCs).
Pulp tissues were obtained from both healthy and carious teeth (n = 5, each) to generate primary cell lines using the explant culture technique. Cell cultures studies were undertaken by generating growth curves, a colony forming unit and a mineralization assay analysis. The expression of vimentin was assessed using immunocytochemistry (ICC), and the gene expression of above-mentioned genes was determined using quantitative real-time reverse-transcription polymerase chain reaction.
ncHDPCs and cHDPCs were successfully isolated and cultured from healthy and inflamed human dental pulp tissue. At passage 4, both HDPC types demonstrated a typical spindle morphology with positive vimentin expression. No statistical difference was observed between ncHDPCs and cHDPCs in their growth characteristics or ability to differentiate into a mineralizing phenotype. ncHDPCs showed a statistically significant higher colony forming efficiency than cHDPCs. The gene expression levels of TLR-2, TLR-4, TLR-9, TNF-α, IL-6, IL-8, IL-17R, IL-23A, NF-κB, MAPK1, DMP1, DSPP and SOX2 were significantly higher in cHDPCs compared with ncHDPC cultures.
cHDPCs retain their differentiation potential and inflammatory phenotype in vitro. The inflamed tooth pulp contains viable stem/progenitor cell populations which have the potential for expansion, proliferation and differentiation into a mineralizing lineage, similar to cells obtained from healthy pulp tissue. These findings have positive implications for regenerative endodontic procedures.
本研究旨在从龋病牙(cHDPCs)中分离和培养人牙髓细胞,并比较其生长特性、集落形成效率、矿化潜能以及 Toll 样受体(TLR)-2、TLR-4、TLR-9、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6、IL-8、IL-17A、IL-17R、IL-23A、核因子-κB(NF-κB)、丝裂原活化蛋白激酶(MAPK1)、牙本质基质蛋白(DMP)-1、牙本质涎磷蛋白(DSPP)、性决定区 Y 框 2(SOX2)和增殖标志物 Ki-67(MKi67)的基因表达,与从健康或非龋病牙(ncHDPCs)中分离的细胞进行比较。
使用组织块培养技术从健康和龋病牙(n = 5,每个)中获取牙髓组织,以生成原代细胞系。通过生成生长曲线、集落形成单位和矿化分析进行细胞培养研究。使用免疫细胞化学(ICC)评估波形蛋白的表达,并用定量实时逆转录聚合酶链反应确定上述基因的基因表达。
成功地从健康和炎症性人牙髓组织中分离和培养了 ncHDPCs 和 cHDPCs。在第 4 代时,两种 HDPC 类型均表现出典型的纺锤形形态,波形蛋白表达阳性。在生长特性或分化为矿化表型的能力方面,ncHDPCs 和 cHDPCs 之间没有统计学差异。ncHDPCs 的集落形成效率明显高于 cHDPCs。cHDPC 培养物中 TLR-2、TLR-4、TLR-9、TNF-α、IL-6、IL-8、IL-17R、IL-23A、NF-κB、MAPK1、DMP1、DSPP 和 SOX2 的基因表达水平明显高于 ncHDPC 培养物。
cHDPCs 在体外保留其分化潜能和炎症表型。发炎的牙髓含有有活力的干细胞/祖细胞群体,这些细胞具有扩增、增殖和分化为矿化谱系的潜力,类似于从健康牙髓组织中获得的细胞。这些发现对再生性牙髓治疗有积极意义。