Inhalation Toxicity Research Center, Occupational Safety and Health Research Institute, Korea Occupational Safety and Health Agency, 30, Expro-ro 339 beon-gil, Yuseong-gu, Daejeon, Republic of Korea.
Sci Rep. 2024 Oct 14;14(1):23981. doi: 10.1038/s41598-024-75491-1.
Recently, there have been reports of sarcoma occurring in a Korean science teachers who used a 3D printer with acrylonitrile butadiene styrene (ABS) and polylactic acid (PLA) filaments for educational purposes. However, limited toxicological research data on 3D printing make it challenging to confirm a causal relationship between 3D printing and cancer. Therefore, occupational accidents involving teachers who have developed sarcoma have not been officially recognized. To address this gap, we aimed to evaluate the carcinogenic potential of particulate matter produced from ABS and PLA filaments commonly used in 3D printing. We created a generator mimicking 3D printing to generate particulate matter, which was used as an experimental material. The collected particulate matter was exposed to an in vitro system to investigate genetic damage, effects on cell transformation, and changes in carcinogenesis-related genes. Various assays, such as the comet assay, cell transformation assays, microarray analysis, and glucose consumption measurement, were employed. Cytotoxicity tests performed to determine the exposure concentration for the comet assay showed that cell viability was 83.6, 62.6, 42.0, and 10.2% for ABS at exposure concentrations of 50, 100, 200, and 400 µg/mL, respectively. PLA showed 91.7, 80.3, 65.1, and 60.8% viability at exposure concentrations of 50, 100, 200, and 400 µg/mL, respectively. Therefore, 50 µg/mL was set as the highest concentration for both ABS and PLA, and 25 and 12.5 µg/mL were set as the medium and low concentrations, respectively. The comet assay showed no changes in genetic damage caused by the particulate matter. Cytotoxicity results performed to establish exposure concentrations in the transformation assay showed that ABS showed cell viability of 88.0, 77.4, 84.7, and 85.5% at concentrations of 1.25, 2.5, 5, and 10 µg/mL, respectively, but few cells survived at concentrations above 20 µg/mL. PLA showed minimal cytotoxicity up to a concentration of 20 µg/ml. Therefore, in the cell transformation assay, a concentration of 10 µg/mL for ABS and 20 µg/mL for PLA was set as the highest exposure concentration, followed by medium and low exposure concentrations with a common ratio of 2. In cell transformation assays, only one transformed focus each was observed for both ABS and PLA particulate matter-exposed cells. The microarray assay revealed changes in gene expression, with a 41.7% change at 10 µg/mL for ABS and an 18.6% change at 20 µg/mL for PLA compared to the positive control group. Analysis of carcinogenesis-related gene expression changes on days 1, 7, and 25 of the promotion phase revealed that in cells exposed to 5 µg/mL of ABS, RBM3 gene expression increased by 3.66, 3.26, and 3.74 times, respectively, while MPP6 gene expression decreased by 0.33, 0.28, and 0.38 times, respectively, compared to the negative control group. Additionally, the measurement of glucose consumption showed that it increased in cells exposed to ABS and PLA particulate matter. Our findings suggest that the carcinogenic potential of ABS- and PLA-derived particulate matter in 3D printing cannot be completely ruled out. Therefore, further research in other test systems and analysis of additional parameters related to carcinogenesis, are deemed necessary to evaluate the carcinogenic risk of 3D printers using these materials.
最近，有报道称，一名韩国科学教师在使用丙烯腈丁二烯苯乙烯（ABS）和聚乳酸（PLA）长丝进行教育目的的 3D 打印时患上了肉瘤。然而，3D 打印的毒理学研究数据有限，难以确定 3D 打印与癌症之间的因果关系。因此，涉及已发展为肉瘤的教师的职业事故尚未得到正式承认。为了解决这一差距，我们旨在评估 ABS 和 PLA 长丝中常见的 3D 打印产生的颗粒物质的致癌潜力。我们创建了一个模拟 3D 打印的发生器来产生颗粒物质，将其作为实验材料。收集的颗粒物质暴露于体外系统中，以研究遗传损伤、对细胞转化的影响以及与致癌相关的基因变化。进行了各种测定，如彗星试验、细胞转化试验、微阵列分析和葡萄糖消耗测量。为确定彗星试验的暴露浓度而进行的细胞毒性试验表明，ABS 在暴露浓度为 50、100、200 和 400 µg/mL 时，细胞活力分别为 83.6、62.6、42.0 和 10.2%。PLA 在暴露浓度为 50、100、200 和 400 µg/mL 时，细胞活力分别为 91.7、80.3、65.1 和 60.8%。因此，将 50 µg/mL 设定为 ABS 和 PLA 的最高浓度，将 25 和 12.5 µg/mL 设定为中低浓度。彗星试验显示颗粒物质未引起遗传损伤的变化。为建立转化试验中的暴露浓度而进行的细胞毒性试验结果表明，ABS 在浓度为 1.25、2.5、5 和 10 µg/mL 时，细胞活力分别为 88.0、77.4、84.7 和 85.5%，但浓度高于 20 µg/mL 时，细胞存活很少。PLA 直至 20 µg/ml 时表现出最小的细胞毒性。因此，在细胞转化试验中，将 ABS 的 10 µg/mL 和 PLA 的 20 µg/mL 设定为最高暴露浓度，随后设定共同比例为 2 的中低暴露浓度。在细胞转化试验中，仅观察到 ABS 和 PLA 颗粒物质暴露细胞的每个转化焦点各一个。微阵列试验显示基因表达发生变化，ABS 为 10 µg/mL 时变化 41.7%，PLA 为 20 µg/mL 时变化 18.6%，与阳性对照组相比。在促进阶段第 1、7 和 25 天对致癌相关基因表达变化的分析表明，在暴露于 5 µg/mL ABS 的细胞中，RBM3 基因表达分别增加了 3.66、3.26 和 3.74 倍，而 MPP6 基因表达分别降低了 0.33、0.28 和 0.38 倍，与阴性对照组相比。此外，葡萄糖消耗的测量表明 ABS 和 PLA 颗粒物质暴露的细胞中葡萄糖消耗增加。我们的研究结果表明，3D 打印中 ABS 和 PLA 衍生颗粒物质的致癌潜力不能完全排除。因此，有必要在其他测试系统中进行进一步的研究，并分析与致癌相关的其他参数，以评估使用这些材料的 3D 打印机的致癌风险。
