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牙本质预处理后生长因子释放和牙髓干细胞附着的体外研究。

Growth factor release and dental pulp stem cell attachment following dentine conditioning: An in vitro study.

机构信息

School of Dentistry, College of Biomedical and Life Sciences, Cardiff University, Cardiff, UK.

College of Dentistry, Taif University, Ta'if, Saudi Arabia.

出版信息

Int Endod J. 2022 Aug;55(8):858-869. doi: 10.1111/iej.13781. Epub 2022 Jun 16.

Abstract

AIM

The aim of the study was to investigate the effect of dentine conditioning agents on growth factor liberation and settlement of dental pulp progenitor cells (DPSCs) on dentine surfaces.

METHODOLOGY

The agents used included ethylenediaminetetraacetic acid (EDTA; 10%, pH 7.2), phosphoric acid (37%, pH < 1), citric acid (10%, pH 1.5) and polyacrylic acid (25%, pH 3.9). Human dentine slices were conditioned for exaggerated conditioning times of 5 and 10 min, so that the growth factor liberation reached quantifiable levels above the limit of detection of the laboratory methods employed. Transforming growth factor beta-1 (TGF-β1) release and surface exposure were quantified by enzyme-linked immunosorbent assay (ELISA) and immunogold labelling. Scanning electron microscopy (SEM) was used to assess the morphology of cells and coverage by DPSCs cultured on dentine surfaces for 8 days.

RESULTS

After 5-min conditioning of dentine slices, citric acid was the most effective agent for growth factor release into the aqueous environment as measured by ELISA (Mann-Whitney U with Bonferroni correction, p < .01 compared with phosphoric and polyacrylic acid). As well as this, dentine slices treated with phosphoric acid for the same period, displayed significantly less TGF-β1 on the surface compared with the other agents used, as measured by immunogold labelling (MWU with Bonferroni correction, p < .05). After 8 days, widespread coverage by DPSCs on dentine surfaces conditioned with citric acid and EDTA were evident under SEM. On dentine surfaces conditioned with phosphoric and polyacrylic acid, respectively, less spread cells and inconsistent cell coverage were observed.

CONCLUSIONS

Based on the findings of this in vitro study, a desirable biological growth factor-mediated effect may be gained when conditioning dentine by milder acidic or chelating agents such as citric acid and EDTA. The results must be interpreted in the context that the potential of the applied materials inducing a desirable biological response in DPSCs is only one consideration amongst other important ones in a clinical setting. However, it is crucial to look beyond the mere physical effects of materials and move towards biologically based treatment approaches as far as the restorative management of teeth with viable dental pulps are concerned.

摘要

目的

本研究旨在探讨牙本质处理剂对生长因子释放和牙髓祖细胞(DPSCs)在牙本质表面附着的影响。

方法

所用的处理剂包括乙二胺四乙酸(EDTA;10%,pH7.2)、磷酸(37%,pH<1)、柠檬酸(10%,pH1.5)和聚丙烯酸(25%,pH3.9)。对人牙本质片进行过度处理,处理时间分别为 5 分钟和 10 分钟,以使生长因子释放达到实验室方法检测限以上的可量化水平。通过酶联免疫吸附测定(ELISA)和免疫胶体金标记定量测定转化生长因子-β1(TGF-β1)的释放和表面暴露。扫描电子显微镜(SEM)用于评估细胞形态和在牙本质表面培养 8 天后 DPSCs 的覆盖情况。

结果

牙本质片经 5 分钟处理后,柠檬酸作为 ELISA 测量的最有效的生长因子释放到水性环境中的试剂(曼-惠特尼 U 检验,Bonferroni 校正,p<0.01 与磷酸和聚丙烯酸相比)。此外,磷酸处理相同时间的牙本质片,与其他使用的试剂相比,表面上的 TGF-β1 明显较少,免疫胶体金标记测量(MWU 检验,Bonferroni 校正,p<0.05)。8 天后,SEM 显示,在柠檬酸和 EDTA 处理的牙本质表面上,DPSCs 广泛覆盖。在磷酸和聚丙烯酸处理的牙本质表面上,观察到细胞扩散较少,细胞覆盖不一致。

结论

根据本体外研究的结果,当通过柠檬酸和 EDTA 等较温和的酸性或螯合剂处理牙本质时,可能获得理想的生物生长因子介导的效果。必须在应用材料在 DPSCs 中诱导理想的生物学反应的潜力只是临床环境中其他重要考虑因素之一的背景下解释这些结果。然而,在涉及有活力牙髓的牙齿的修复管理时,必须超越材料的纯粹物理效应,朝着基于生物学的治疗方法发展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b85/9541952/b773ee0234c8/IEJ-55-858-g003.jpg

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