Development of Biological Safty Testing Using Endocrine Disruping Effects

最近，与内分泌干扰物有关的公共问题一直在增加。作为一种内分泌干扰物，双酚A(BPA)是合成Bis-GMA的原料，Bis-GMA用于牙科材料，如裂隙封闭剂、复合树脂和粘结剂，聚碳酸酯用于牙科材料，如正畸托槽、临时牙冠和牙齿树脂。在牙科领域，迫切需要对聚合物材料中化学物质的安全性进行评价，明确牙科材料中残留的双酚A，分析双酚A在牙科材料中的洗脱规律，建立灵敏的内分泌双酚A检测系统。本研究的目的是弄清BPA等内分泌干扰物是否被从牙科材料中滤出，并发展牙科材料的洗脱测试，以及利用乳腺癌细胞株阐明BPA处理引起的基因变化。聚合材料…中可浸出的双酚A水平用带有荧光检测器的高效液相色谱仪对浸泡在人工唾液中的更多S类封闭剂、复合树脂和粘结剂的检测下限(0.5 ng/mL，即5.7-15.8ngBPA/g树脂)。另一方面，从浸泡在人工唾液中的正畸托槽、临时全冠和树脂牙中检测到可浸出的双酚A。此外，浸泡在人唾液中的正畸托槽中的双酚A和对叔丁基苯酚(t-BUP)的浸出量比浸入人工唾液中的托槽中的浸出水平更高。与对照组相比，浸泡在人唾液中的残留BPA和t-BUP增加。这些结果表明，在聚合物的洗脱试验中应使用人的唾液作为浸泡溶剂，以反映模拟的口腔环境。在传统的细胞增殖方法中，使用MCF-7来评价内分泌干扰物的雌激素活性。然而，由于该细胞系在细胞培养过程中发生转化，所以使用了T47D细胞系，即乳腺癌细胞系。该细胞重复性好，与MCS-7相比具有很高的灵敏度。经BPA处理24小时后，提取细胞的mRNA，进行差异显示。BPA处理24小时后，NADH脱氢酶亚型表达明显增加。较少

Recently, public problems concernig endocrine disruptors have been increasing. One of endocrine disruptors, bisphenol A(BPA)is a raw material that synthesizes Bis-GMA which is used as dental materials such as fissure sealants, composite resins and bondings as well as that does polycarbonate which is used as dental materials such as orthodontic bracket, temporary crown and teeth resins. In dental field, there are necessary to evaluate safety of chemical substances in polymer materials and it is urgent to clarify the residual BPA in dental materials, to analyze the elution pattern of BPA from it and to develop the sensitive detection system of endocrine disrupors. The purpose of the present study was to clarify whether endocrine disruptors such as BPA were leached from dental materials and to develop the elution testing of dental materials as well as to clarify the changes in genes elicited by BPA-treatment using breast cancer cell lines. Levels of leachable BPA from polymerized material … More s such as fissure sealants, composite resins and bondings immersed in artificial saliva was under detection limit(0.5ng/mL, namely 5.7-15.8ng BPA/g resin)using a high-performance liquid chromatograpy with a fluorescent detector. On the other hand, leachable BPA was detected from orthodontic brackets, temporary crowns and resin teeth immersed in artificial saliva. Furthermore, levels of leaching BPA and p-t-butylphenol(t-BuP)from orthodontic brackets immersed in human saliva was more inreasing than those of orthodontic brackets immersed in artificial saliva. Residual BPA and t-BuP immersed in human saliva was increased compared to controls. These results indicate that human saliva should be used as the immersion solvent for the elution testing of polymers to reflect the simulated oral environment. In conventional cell proliferation methods to evaluate the estrogenecity of the endocrine disruptors, MCF-7 was used. However, as this cell line was transformed during cell culture, T47D cell line, breast cancer cell line, was used. This cell was well reproduced and highly sensitve compared to MCS-7. mRNA was extracted from this cell treated BPA for 24 hours and then differential display was performed. Examining increased cDNA of expression, NADH dehydrogenase subtype was greatly increased after BPA treatment for 24 hours. Less