Biological Effects of New Chemical-Mechanical Caries Removal Products on Human Dental Pulp Stem Cells.

IF 2.9 2区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Caries Research Pub Date : 2024-07-22 DOI:10.1159/000540119

Sergio López-García, Nuria Pérez-Guzmán, Francisco J Rodríguez-Lozano, María Pilar Pecci-Lloret, David García-Bernal, Laura Murcia, Ricardo E Oñate-Sánchez, Carmen Llena

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引用次数: 0

Abstract

Introduction: The aim of this study was to compare the biological effects of four chemical caries removal materials and to assess their cytotoxicity using human dental pulp stem cells (hDPSCs).

Methods: The products evaluated are: 1 - papain-based product (BRIX 3000®); 2 - papain/chloramine based products (NATURAL-CARE and Papacárie Duo®); and 3 - chloramine based product (Cariesolut). The following in vitro experiments were carried out: IC50 measurement, cell metabolic activity (MTT) assay, cell migration, immunofluorescence experiment, cell apoptosis analysis, and reactive oxygen species (ROS) production analysis. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p < 0.05).

Results: The IC50 values were: Brix 3000: 0.596%; Papacárie Duo: 0.052%; NATURAL CARE: 1.034%; and Cariesolut: 0.020%. The MTT assays showed non-adequate cell viability of all chemical-mechanical caries removal tested at 2% at 24, 48, and 72 h (p < 0.001). The same behaviour was observed at 0.1% in the Papacárie Duo and Cariesolut groups. In contrast, 0.1% of Brix 3000 at all times and NATURAL CARE at 24 h treated cells showed cell viability rates similar to the control group. At 0.01% only Brix 3000 did not show statistically significant differences at any time. Delayed cell migration was observed in all hDPSCs treated with Papacárie Duo and Cariesolut (p < 0.01 and p < 0.001). Phalloidin staining images showed a high confluence of cells in the presence of NATURAL CARE, similar to the control group. On the contrary, no cells were observed in Brix 3000 and Cariesolut at 2% and 0.1% concentrations. Papacárie Duo showed cells at all concentrations, but hDPSCs treated at 0.01% concentration exhibited better proliferation and spreading than those in the control group. Apoptosis essay showed that Brix 3000 at both 0.1% and 0.01% had a percentage of live cells higher than 99%, with 68.4% live cells at 2%, 3.69% early apoptotic cells, and 27.9% late apoptotic cells. Conversely, the rest of the materials showed an abundance of apoptotic cells, even at low concentrations. 0.1% and 0.01% of BRIX 3000 did not affect the ROS production levels, while 2% of BRIX 3000 counterpart very significantly increased the percentage of CM-H2DCFDA positive cells. Again, all concentrations of Cariesolut showed significantly higher levels of ROS production than those observed in control cells.

Conclusion: Our results suggest that Brix 3000 would be the most suitable material for chemical caries removal, with Papacárie Duo and NATURAL CARE also being good options, and discourage the use of Cariesolut due to its low cytocompatibility on dental pulp stem cells.

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新型化学机械除龋产品对人类牙髓干细胞的生物效应。

目的：本研究旨在比较四种化学除龋材料的生物效应，并利用人体牙髓干细胞（hDPSCs）评估其细胞毒性：评估的产品有1 -木瓜蛋白酶类产品（BRIX 3000®）；2 -木瓜蛋白酶/氯胺类产品（NATURAL-CARE 和 Papacárie Duo®）；3 -氯胺类产品（Cariesolut）：进行了以下体外实验：IC50测定、细胞代谢活性（MTT）测定、细胞迁移、免疫荧光实验、细胞凋亡分析和活性氧（ROS）产生分析。统计分析采用单因素方差分析和 Tukey 后检验（p<0.05）：IC50 值为结果：IC50 值分别为：Brix 3000：0.596%；Papacárie Duo：0.052%；NATURAL CARE：1.034%；Cariesolut：0.020%：0.020%.MTT 检测显示，在 24、48 和 72 小时内，所有测试的 2% CMCR 细胞存活率均不达标（p<0.001）。Papacárie Duo 和 Cariesolut 组在 0.1% 的浓度下也出现了同样的情况。相比之下，0.1% 的 Brix 3000 和 NATURAL CARE 在 24 小时内处理的细胞存活率与对照组相似。在 0.01% 的浓度下，只有 Brix 3000 在任何时间都没有显示出显著的统计学差异。用 Papacárie Duo 和 Cariesolut 处理的所有 hDPSCs 都观察到细胞迁移延迟（p<0.01 和 p<0.001）。类木磷脂染色图像显示，细胞在 NATURAL CARE 的作用下高度融合，与对照组相似。相反，在浓度为 2% 和 0.1% 的 Brix3000 和 Cariesolut 中没有观察到细胞。Papacárie Duo 在所有浓度下都出现了细胞，但与对照组相比，在 0.01% 浓度下处理的 hDPSCs 表现出更好的增殖和扩散能力。细胞凋亡检测显示，0.1% 和 0.01% 浓度的 Brix 3000 活细胞比例高于 99%，2% 浓度的活细胞比例为 68.4%，早期凋亡细胞比例为 3.69%，晚期凋亡细胞比例为 27.9%。相反，其余材料即使在低浓度下也显示出大量凋亡细胞。0.1% 和 0.01% 的 BRIX 3000 不会影响 ROS 生成水平，而 2% 的 BRIX 3000 则会显著增加 CM-H2DCFDA 阳性细胞的比例。同样，所有浓度的 Cariesolut 所产生的 ROS 水平都明显高于对照细胞：我们的研究结果表明，Brix 3000 是最适合用于化学除龋的材料。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Caries Research 医学-牙科与口腔外科

CiteScore

6.30

自引率

7.10%

发文量

审稿时长

6-12 weeks

期刊介绍： ''Caries Research'' publishes epidemiological, clinical and laboratory studies in dental caries, erosion and related dental diseases. Some studies build on the considerable advances already made in caries prevention, e.g. through fluoride application. Some aim to improve understanding of the increasingly important problem of dental erosion and the associated tooth wear process. Others monitor the changing pattern of caries in different populations, explore improved methods of diagnosis or evaluate methods of prevention or treatment. The broad coverage of current research has given the journal an international reputation as an indispensable source for both basic scientists and clinicians engaged in understanding, investigating and preventing dental disease.