Department of Preventive Medicine and Public Health, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan.
J Occup Health. 2010;52(2):106-14. doi: 10.1539/joh.l9129. Epub 2010 Feb 3.
We performed a cytotoxicity study by cell magnetometry, measured lactate dehydrogenase (LDH) activity by enzyme assay, detected DNA ladder formation, and performed morphological examination by electron microscopy in order to evaluate the safety of high temperature wool (HT wool), an asbestos substitute, using long and short chrysotile fibers (CF) as positive controls and phosphate buffered saline (PBS) as a negative control.
Alveolar macrophages were isolated from male Fisher rats. Following the addition of iron oxide particles (Fe(3)O(4)) to macrophages, HT wool, long or short CF was added. Then, the remanence strength was measured for 20 min after magnetization by an external field. Percent LDH release was calculated after determining LDH activity. DNA was detected using an apoptosis detection kit. Morphological observation was performed by taking electron micrographs of macrophages in the groups treated with HT wool and long- and short-CF.
Rapid relaxation, an indicator of decay of cytotoxicity, was observed by cell magnetometry immediately after magnetization was ended in the groups treated with HT wool and PBS, showing that HT wool causes no harmful effect on the cytoskeleton. The CF-treated groups had higher LDH activity than the PBS- and HT wool-treated groups. No fragmentation of DNA was observed in any group. In morphological observation, cytotoxicity in macrophages was lower in the HT wool-treated groups than in the CF-treated groups.
The results suggest that HT wool has no cytotoxicity, as evaluated by cell magnetometry, enzyme assay, DNA ladder detection and morphological examination.
通过细胞磁力计进行细胞毒性研究，通过酶测定法测量乳酸脱氢酶（LDH）活性，检测 DNA 梯形成，并通过电子显微镜进行形态学检查，以评估高温羊毛（HT 羊毛）的安全性，将石棉替代品的长、短温石棉纤维（CF）作为阳性对照，磷酸盐缓冲盐水（PBS）作为阴性对照。
从雄性 Fisher 大鼠中分离肺泡巨噬细胞。在向巨噬细胞中添加氧化铁颗粒（Fe(3)O(4))后，添加 HT 羊毛、长或短 CF。然后，在外磁场磁化后测量 20 分钟的剩余强度。测定 LDH 活性后计算 LDH 释放的百分比。使用凋亡检测试剂盒检测 DNA。通过拍摄用 HT 羊毛和长、短-CF 处理的巨噬细胞组的电子显微镜照片进行形态学观察。
细胞磁力计观察到，HT 羊毛和 PBS 处理组在磁化结束后立即出现快速弛豫，这是细胞毒性衰减的指标，表明 HT 羊毛对细胞骨架没有有害影响。CF 处理组的 LDH 活性高于 PBS 和 HT 羊毛处理组。任何组都未观察到 DNA 片段化。在形态学观察中，HT 羊毛处理组的巨噬细胞细胞毒性低于 CF 处理组。
细胞磁力计、酶测定法、DNA 梯检测和形态学检查评估结果表明，HT 羊毛没有细胞毒性。
