Growth factor release and dental pulp stem cell attachment following dentine conditioning: An in vitro study.
Growth factor release and dental pulp stem cell attachment following dentine conditioning: An in vitro study.
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DOI：
10.1111/iej.13781
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发表时间：
2022-08
影响因子：
5
通讯作者：
Sloan, Alastair J.
中科院分区：
文献类型：
--
作者：
Sadaghiani, Leili;Alshumrani, Abdullah M.;Gleeson, Hannah B.;Ayre, Wayne Nishio;Sloan, Alastair J.
本研究旨在探讨牙本质调理剂对牙髓祖细胞（DPSC）在牙本质表面生长因子释放和定居的影响。使用的试剂包括乙二胺四乙酸（EDTA; 10%，pH 7.2）、磷酸（37%，pH < 1）、柠檬酸（10%，pH 1.5）和聚丙烯酸（25%，pH 3.9）。将人牙本质切片调节5和10分钟的夸张调节时间，使得生长因子释放达到高于所采用的实验室方法的检测限的可定量水平。通过酶联免疫吸附试验（ELISA）和免疫金标记对转化生长因子β 1（TGF-β1）释放和表面暴露进行定量。用扫描电镜观察DPSC在牙本质表面培养8天后的细胞形态和覆盖情况。牙本质切片处理5分钟后，通过ELISA测定，柠檬酸是将生长因子释放到水环境中的最有效试剂（采用Bonferroni校正的Mann-Whitney U，与磷酸和聚丙烯酸相比p <0.01）。除此之外，用磷酸处理相同时间的牙本质切片，与使用的其他试剂相比，表面上显示出显著更少的TGF-β1，如通过免疫金标记所测量的（MWU与Bonferroni校正，p <0.05）。8天后，在SEM下，用柠檬酸和EDTA处理的牙本质表面上的DPSC的广泛覆盖是明显的。在分别用磷酸和聚丙烯酸处理的牙本质表面上，观察到较少的扩散细胞和不一致的细胞覆盖。根据这项体外研究的结果，当使用弱酸性或螯合剂（如柠檬酸和EDTA）调节牙本质时，可能会获得理想的生物生长因子介导的效果。结果必须在这样的背景下解释，即所应用的材料在DPSC中诱导期望的生物反应的潜力仅是临床环境中其他重要因素中的一个考虑因素。然而，至关重要的是，要超越材料的单纯物理效应，并转向基于生物学的治疗方法，就有活力牙髓的牙齿的修复管理而言。
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. 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. 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. 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.
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