Department of Stomatology, Kunming General Hospital of Chengdu Military Command, Teaching Hospital of Kunming Medical University, Kunming, China.
Department of Stomatology, Kunming Municipal Hospital of Traditional Chinese Medicine, Kunming, China.
Int Endod J. 2018 Feb;51 Suppl 2:e157-e166. doi: 10.1111/iej.12812. Epub 2017 Aug 1.
To assess the effects of 2-hydroxyethyl methacrylate (HEMA) on proliferation and migration of human pulp cells, as well as on matrix metalloproteinase (MMP-2 and MMP-9) expression in human odontoblast-like cells, contributing to the goal of determining the relationship between resin materials and MMP activity in pulp-dentine complexes.
Dental pulp cell cultures were established from pulp tissue of human teeth extracted for orthodontic purposes. Pulp cell differentiation was characterized in the presence of dentine sialophosphoprotein, bone sialoprotein and alkaline phosphatase by reverse transcription polymerase chain reaction. MMP activity was assessed by gelatine zymography with media containing HEMA. Cell viability was evaluated using methyl thiazolyl tetrazolium assay for 24-72 h. Cell migration was tested using Transwell migration assay. Western blotting was used to visualize MMP expression with the nontoxic HEMA concentrations (0-400 μg mL ) for 48 h.
Pulp cell proliferation decreased with HEMA exposure in a time- and concentration-dependent manner. HEMA concentrations ≤400 μg mL did not induce changes in cell viability at 48 h (P < 0.05). Pulp cells were induced to differentiate into odontoblast-like cells in media containing 5 mg mL ascorbic acid and 10 mmol L β-sodium glycerophosphate for 3-4 weeks. After incubation with HEMA, dose-dependent inhibition was observed; HEMA had a strong inhibitory effect on MMP activity. Compared with the control group, cell migration and MMP expression were inhibited significantly with increasing HEMA concentration at noncytotoxic doses (P < 0.05).
Cell viability was not affected at HEMA concentrations ≤400 μg mL . Within this range, HEMA inhibited MMP-2 and MMP-9 expression and activity, which may protect against type I collagen degradation effectively during dentine adhesive procedures.
评估 2-羟乙基甲基丙烯酸酯（HEMA）对人牙髓细胞增殖和迁移的影响，以及对人成牙本质细胞样细胞基质金属蛋白酶（MMP-2 和 MMP-9）表达的影响，以期确定树脂材料与牙髓牙本质复合体中 MMP 活性之间的关系。
从因正畸需要而拔除的人牙齿牙髓组织中建立牙髓细胞培养物。通过逆转录聚合酶链反应，用牙本质涎磷蛋白、骨涎磷蛋白和碱性磷酸酶来鉴定牙髓细胞分化。通过含 HEMA 的凝胶酶谱法评估 MMP 活性。用噻唑蓝比色法在 24-72 小时评估细胞活力。通过 Transwell 迁移试验检测细胞迁移。用 Western blot 法在 48 小时内用无毒的 HEMA 浓度（0-400μg/mL）可视化 MMP 表达。
HEMA 暴露会导致牙髓细胞增殖随时间和浓度依赖性降低。HEMA 浓度≤400μg/mL 时，48 小时内细胞活力无变化（P<0.05）。在含有 5mg/mL 抗坏血酸和 10mmol/L β-甘油磷酸钠的培养基中培养 3-4 周，可诱导牙髓细胞分化为成牙本质细胞样细胞。孵育 HEMA 后，观察到剂量依赖性抑制作用；HEMA 对 MMP 活性有强烈的抑制作用。与对照组相比，在非细胞毒性剂量下，随着 HEMA 浓度的增加，细胞迁移和 MMP 表达显著受到抑制（P<0.05）。
在 HEMA 浓度≤400μg/mL 时，细胞活力不受影响。在此范围内，HEMA 抑制 MMP-2 和 MMP-9 的表达和活性，这可能在牙本质粘接过程中有效防止 I 型胶原降解。
