In Vivo Molecular Toxicity Profile of Dental Bioceramics in Embryonic Zebrafish (Danio rerio)

The investigation of the biocompatibility of potential and commercially available dental material is a major challenge in dental science. This study demonstrates that the zebrafish model is a novel in vivo model for investigating the biocompatibility of dental materials. Two commercially available dental materials, mineral trioxide aggregate (MTA) and Biodentine, were assessed for their biocompatibility. The biocompatibility analysis was performed in embryonic zebrafish with the help of standard toxicity assays measuring essential parameters such as survivability and hatching. The mechanistic and comparative analysis of toxicity was performed by oxidative stress analysis by measuring ROS induction and apoptosis in zebrafish exposed to dental materials at different concentrations. The molecular investigation at the protein level was done by a computational approach using in silico molecular docking and pathway analysis. The toxicity analysis showed a significant reduction in hatching and survivability rates along with morphological malformations with an increase in the concentration of exposed materials. ROS and apoptosis assay results revealed a greater biocompatibility of Biodentine as compared to that of MTA which was concentration-dependent. In silico analysis showed the significant role of the tricalcium silicate–protein (Sod1, tp53, RUNX2B) interaction in an exhibition of toxicity. The study provides a new vision and standard in dental material sciences for assessing the biocompatibility of potential novel and commercially available dental materials.潜在及商业化牙科材料的生物相容性研究，是牙科科学领域的核心挑战之一。本研究证实，斑马鱼模型可作为一种新型体内模型，用于牙科材料的生物相容性评估。本研究选取两款商业化牙科材料——矿物三氧化物凝聚体（mineral trioxide aggregate, MTA）与Biodentine，对其生物相容性开展评估。实验以胚胎斑马鱼为对象，通过检测存活率、孵化率等关键参数的标准毒性实验，完成生物相容性分析。针对毒性的机制与对比研究，则通过氧化应激分析实现：即检测不同浓度牙科材料暴露下斑马鱼体内的活性氧（reactive oxygen species, ROS）诱导水平与细胞凋亡情况。本研究采用计算机模拟（in silico）分子对接与通路分析的计算方法，完成蛋白质层面的分子探究。毒性分析结果显示，随着暴露材料浓度升高，斑马鱼的孵化率与存活率显著下降，同时伴随形态畸形。活性氧与细胞凋亡检测结果表明，Biodentine的生物相容性优于MTA，且该差异呈浓度依赖性。计算机模拟分析显示，硅酸三钙与蛋白质（Sod1、tp53、RUNX2B）的相互作用在毒性表现中发挥关键作用。本研究为牙科材料科学领域中潜在新型及商业化牙科材料的生物相容性评估工作，提供了全新的研究视角与标准范式。