Cho Tae-Ho, Lee Jin-Woo, Lee Mu-Hyoung
Department of Dermatology, College of Medicine, Kyunghee University, Seoul, Korea.
Photodermatol Photoimmunol Photomed. 2008 Jun;24(3):110-4. doi: 10.1111/j.1600-0781.2008.00347.x.
No comparative and simultaneous in vitro studies have been performed to determine the cytotoxic dose of narrowband UVB (NBUVB) and broadband UVB (BBUVB) for keratinocytes, melanocytes, and fibroblasts. Culture medium was often replaced with phosphate-buffered saline (PBS) before UV irradiation; however, its amount differed across studies. We determined the cytotoxic doses of NBUVB and BBUVB and tested for changes in viability according to the amount of PBS.
We exposed cultured human keratinocytes, melanocytes, and fibroblasts to ultraviolet light in the range 12.5-1000 mJ/cm(2) for NBUVB and 1.25-100 mJ/cm(2) for BBUVB. The viability was assessed after 24 h. We also determined changes in viability at cytotoxic doses according to the amount of PBS (40, 80, and 120 microl/well in a 96-well plate).
Cytotoxicity was observed at doses of 100, 200, and 400 mJ/cm(2) for NBUVB and 5, 10, and 25 mJ/cm(2) for BBUVB in keratinocytes, melanocytes, and fibroblasts, respectively. At cytotoxic doses, there was no change in viability according to the amount of PBS.
Fibroblasts are more resistant to UVB irradiation, irrespective of the amount of NBUVB and BBUVB, than keratinocytes and melanocytes. The amount of PBS during irradiation had no effect on viability.
尚未进行比较性的同时体外研究来确定窄谱中波紫外线(NBUVB)和宽谱中波紫外线(BBUVB)对角质形成细胞、黑素细胞和成纤维细胞的细胞毒性剂量。紫外线照射前,培养基常被磷酸盐缓冲盐水(PBS)替代;然而,不同研究中其用量有所不同。我们测定了NBUVB和BBUVB的细胞毒性剂量,并根据PBS用量测试了细胞活力的变化。
我们将培养的人角质形成细胞、黑素细胞和成纤维细胞分别暴露于NBUVB 12.5 - 1000 mJ/cm²和BBUVB 1.25 - 100 mJ/cm²范围内的紫外线。24小时后评估细胞活力。我们还根据PBS用量(96孔板中每孔40、80和120微升)测定细胞毒性剂量下细胞活力的变化。
在角质形成细胞、黑素细胞和成纤维细胞中,NBUVB剂量为100、200和400 mJ/cm²,BBUVB剂量为5、10和25 mJ/cm²时分别观察到细胞毒性。在细胞毒性剂量下,细胞活力并未因PBS用量而发生变化。
无论NBUVB和BBUVB的剂量如何,成纤维细胞比角质形成细胞和黑素细胞对紫外线照射更具抗性。照射期间PBS的用量对细胞活力没有影响。