Programa de Pós-Graduação em Ciências da Saúde, Universidade de Brasília, Brasília, Distrito Federal, Brazil.
Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, SGAN 916N - Av. W5 - Campus II - Modulo C, Room C-22170.790-160, Brasília, Distrito Federal, Brazil.
Sci Rep. 2023 Jun 12;13(1):9531. doi: 10.1038/s41598-023-36748-3.
Host Defense Peptides (HDPs) have, in previous studies, been demonstrating antimicrobial, anti-inflammatory, and immunomodulatory capacity, important factors in the repair process. Knowing these characteristics, this article aims to evaluate the potential of HDPs IDR1018 and DJK-6 associated with MTA extract in the repair process of human pulp cells. Antibacterial activity of HDPs, MTA and HDPs combined with MTA in Streptococcus mutans planktonic bacteria and antibiofilm activity was evaluated. Cell toxicity was assayed with MTT and cell morphology was observed by scanning electron microscopy (SEM). Proliferation and migration of pulp cells were evaluated by trypan blue and wound healing assay. Inflammatory and mineralization related genes were evaluated by qPCR (IL-6, TNFRSF, DSPP, TGF-β). Alkaline phosphatase, phosphate quantification and alizarin red staining were also verified. The assays were performed in technical and biological triplicate (n = 9). Results were submitted for the calculation of the mean and standard deviation. Then, normality verification by Kolmogorov Smirnov test, analyzing one-way ANOVA. Analyses were considered at a 95% significance level, with a p-value < 0.05. Our study demonstrated that HDPs combined with MTA were able to reduce biofilms performed in 24 h and biofilm performed over 7 days S. mutans biofilm (p < 0.05). IDR1018 and MTA, as well as their combination, down-regulated IL-6 expression (p < 0.05). Tested materials were not cytotoxic to pulp cells. IDR1018 induced high cell proliferation and combined with MTA induced high cellular migration rates in 48 h (p < 0.05). Furthermore, the combination of IDR1018 and MTA also induced high expression levels of DSPP, ALP activity, and the production of calcification nodules. So, IDR-1018 and its combination with MTA could assist in pulp-dentine complex repair process in vitro.
宿主防御肽(HDPs)在以前的研究中表现出抗菌、抗炎和免疫调节能力,这些都是修复过程中的重要因素。了解这些特性,本文旨在评估 IDR1018 和 DJK-6 与 MTA 提取物联合用于人牙髓细胞修复过程的潜力。评估了 HDPs、MTA 和 HDPs 与 MTA 联合对变形链球菌浮游菌的抗菌活性和抗生物膜活性。通过 MTT 测定细胞毒性,通过扫描电子显微镜(SEM)观察细胞形态。通过台盼蓝测定牙髓细胞的增殖和迁移,通过划痕愈合试验测定牙髓细胞的迁移。通过 qPCR(IL-6、TNFRSF、DSPP、TGF-β)评估炎症和矿化相关基因。还验证了碱性磷酸酶、磷酸盐定量和茜素红染色。这些实验在技术和生物学重复(n=9)中进行。结果提交用于计算平均值和标准差。然后,通过 Kolmogorov Smirnov 检验进行正态性验证,分析单因素方差分析。分析在 95%置信水平下进行,p 值<0.05。我们的研究表明,HDPs 与 MTA 联合使用能够减少 24 小时和 7 天的 S. mutans 生物膜形成(p<0.05)。IDR1018 和 MTA 以及它们的组合下调了 IL-6 的表达(p<0.05)。测试材料对牙髓细胞没有细胞毒性。IDR1018 在 48 小时诱导高细胞增殖,与 MTA 联合诱导高细胞迁移率(p<0.05)。此外,IDR1018 与 MTA 的组合还诱导了 DSPP 的高表达水平、ALP 活性和钙化结节的产生。因此,IDR-1018 及其与 MTA 的组合可以辅助体外牙髓牙本质复合体的修复过程。