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富含原花青素的葡萄籽提取物对牙髓细胞的直接和牙本质内生物刺激作用。

Direct and transdentinal biostimulatory effects of grape seed extract rich in proanthocyanidin on pulp cells.

机构信息

Department of Dentistry, Faculty of Health Sciences, University of Brasília, Brasilia, Brazil.

Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, Brasilia, Brazil.

出版信息

Int Endod J. 2019 Apr;52(4):424-438. doi: 10.1111/iej.13019. Epub 2018 Oct 23.

Abstract

AIM

To evaluate the potential biostimulatory effects of grape seed extract (GSE) on a primary culture of human pulp cells.

METHODOLOGY

Human molars were used to obtain the primary pulp cell culture and 0.5-mm dentine discs. For GSE direct exposure, dose-response (0.0065-6.5%) and time response (1-60 min of contact) were examined. For transdentinal exposure, 0.65% of GSE was tested for 24 h. Cellular metabolism, nitric oxide and collagen production, and cell morphology alterations were assessed at periods of 24 and 72 h. After cell differentiation and direct exposure to GSE, the total protein production (TP), alkaline phosphatase activity (ALP) and formation of mineralization nodules (MN) were assessed. The results were analysed by parametric tests or non-parametric tests (α = 0.05).

RESULTS

The lower concentration of GSE tested (0.0065%) was associated with an increase in cellular metabolism, a reduction in the production of nitric oxide and an increase in extracellular matrix synthesis (collagen). Distinct behaviours were observed for the different concentrations, without a reduction of cellular metabolism >10% compared with the control, either when applied directly or transdentinally. SEM revealed no significant change in cell morphology, except for the positive control (H O ). There was no difference in TP, ALP or MN between the control group and the group exposed to GSE.

CONCLUSIONS

Treatment with grape seed extract, even at the highest concentration and longest period, caused neither direct nor transdentinal cytotoxic effects on human pulp cells. Grape seed extract components may play a biostimulatory role and protect dental pulp cells when in direct contact.

摘要

目的

评估葡萄籽提取物(GSE)对人牙髓细胞原代培养的潜在生物刺激作用。

方法

用人磨牙获得原代牙髓细胞培养物和 0.5mm 牙本质片。对于 GSE 的直接暴露,进行了剂量反应(0.0065-6.5%)和时间反应(1-60 分钟接触)的研究。对于跨牙本质暴露,测试了 0.65%的 GSE 24 小时。在 24 小时和 72 小时的不同时期评估细胞代谢、一氧化氮和胶原蛋白的产生以及细胞形态的改变。在细胞分化和直接暴露于 GSE 后,评估总蛋白产生(TP)、碱性磷酸酶活性(ALP)和矿化结节(MN)的形成。使用参数检验或非参数检验(α=0.05)分析结果。

结果

测试的较低浓度的 GSE(0.0065%)与细胞代谢增加、一氧化氮产生减少和细胞外基质合成(胶原蛋白)增加相关。当直接或跨牙本质应用时,观察到不同浓度的不同行为,但与对照相比,细胞代谢没有降低超过 10%。SEM 显示细胞形态没有明显变化,除了阳性对照(H₂O)。TP、ALP 或 MN 在对照组和暴露于 GSE 的组之间没有差异。

结论

即使在最高浓度和最长时间下,葡萄籽提取物的处理也不会对人牙髓细胞造成直接或跨牙本质的细胞毒性作用。葡萄籽提取物的成分在直接接触时可能发挥生物刺激作用并保护牙髓细胞。

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