Objectives

To assess in vitro the effect of experimental mesoporous BAG, on human dental pulp cells (hDPCs) behavior in terms of cytocompatiblity and bioactivity via mineralization potential.

Methods

Fine (FP) and large particles (LP) of a fixed BAG composition named 0NaMBG have been elaborated by a sol-gel process. In vitro assessment was achieved on cultured primary hDPCs using indirect contact. The effect of the concentration of 800 μg/mL on cell metabolic activity and cytotoxicity were examined by performing Alamar blue and crystal violet assays. Alizarin Red staining was used to detect and quantify the formation of mineralized nodules and ALP activity was colourimetrically quantified. The expression of Odontogenic markers: DMP-1 and osteopontin (OPN) expression and cell morphology was evaluated by confocal microscopy.

Results

According to the Alamar blue and crystal violet assay, 0NaMBG samples were non-cytotoxic. Cells treated with 0NaMBG particles expressed higher metabolic activity than control cells, especially for LP. Both FP and LP significantly increased both extra and intra cellular ALP activity. hDPCs exhibited good cell spreading and adhesion in the presence of FP and LP extracts by confocal imaging. Further, Alizarin red S assay demonstrated more mineralization nodules and significant enhancement of the extracellular calcium deposition when cells were interfaced with both FP and LP compared to the control cells. Moreover, LP extracts enhanced the production and secretion of odontogenic markers: dentin matrix protein 1 (DMP-1) and osteopontin.

Significance

LP have a higher surface area and pore volume, which could explain their greater bioactivity in contact with pulp cells. The clinical relevance of these findings implicate that 0NaMBG could be used as fillers in dental therapeutic materials suitable for dentin and/or pulp tissues preservation.

中文翻译：

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实验性生物活性玻璃对牙髓细胞体外行为的影响。

目标

为了评估在体外实验中孔袋的效果，在cytocompatiblity和生物活性方面对人牙髓细胞（hDPCs）行为通过矿化的潜力。

方法

固定的BAG组合物名为0NaMBG的细颗粒（FP）和大颗粒（LP）已通过溶胶-凝胶工艺进行了精细加工。使用间接接触在培养的原代hDPC上进行了体外评估。通过进行Alamar蓝和结晶紫测定，检测了800μg/ mL浓度对细胞代谢活性和细胞毒性的影响。茜素红染色用于检测和定量矿化结节的形成，并用比色法定量ALP活性。通过共聚焦显微镜评估成牙标记物：DMP-1和骨桥蛋白（OPN）的表达以及细胞形态。

结果

根据Alamar蓝和结晶紫测定，0NaMBG样品无细胞毒性。用0NaMBG颗粒处理的细胞比对照细胞表达更高的代谢活性，特别是对于LP。FP和LP均显着增加细胞外和细胞内ALP活性。通过共聚焦成像，在FP和LP提取物的存在下，hDPCs表现出良好的细胞扩散和粘附性。此外，与对照细胞相比，当细胞与FP和LP接触时，茜素红S分析显示出更多的矿化结节和细胞外钙沉积的显着增强。此外，LP提取物增强了牙源性标记物（牙本质基质蛋白1（DMP-1）和骨桥蛋白）的产生和分泌。

意义

LP具有较高的表面积和孔体积，这可以解释其与纸浆细胞接触时具有更高的生物活性。这些发现的临床意义暗示，0NaMBG可用作适合牙本质和/或牙髓组织保存的牙科治疗材料中的填充剂。