Apoptotic cell death induced by 5-aminolaevulinic acid-mediated photodynamic therapy of hypertrophic scar-derived fibroblasts
Apoptotic cell death induced by 5-aminolaevulinic acid-mediated photodynamic therapy of hypertrophic scar-derived fibroblasts
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DOI：
10.3109/09546634.2012.697987
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发表时间：
2014-10-01
影响因子：
2.9
通讯作者：
Li, Wei
中科院分区：
文献类型：
--
作者：
Li, Xin;Zhou, Zhao Pin;Li, Wei
目的：探讨5-氨基乙酰丙酸（ALA）光动力疗法（PDT）对增生性瘢痕（HS）真皮成纤维细胞的作用。方法：HS样本来自5名接受手术的患者，健康供体的正常皮肤作为对照。分离真皮成纤维细胞，并与不同浓度的ALA一起培养6小时。用共聚焦显微镜观察细胞内原卟啉IX（PpIX）的表达。ALA处理5小时后，用红色激光（635 nm波长）以10 mW/cm（2）的功率密度和0.5 - 4 J/cm（2）的能量密度照射细胞。24 h后用CCK-8试剂盒测定细胞存活率。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记和流式细胞术分析annexin V检测细胞死亡。结果：ALA治疗后，HS患者和健康供体的成纤维细胞中观察到细胞内PpIX积累。激光照射后，两种细胞类型中的活细胞以剂量依赖性方式随能量密度而减少。此外，低剂量PDT观察到凋亡细胞死亡，而高剂量PDT增加坏死细胞。结论：在ALA处理后，HS衍生的成纤维细胞有效地积累PpIX，并且可以通过受控激光照射通过细胞凋亡来消除。
Objective: To investigate the effects of 5-aminolevulinic acid (ALA)-mediated photodynamic therapy (PDT) on dermal fibroblasts from hypertrophic scars (HSs). Methods: HS samples were obtained from five patients who underwent surgery, and normal skin from healthy donors was used as a control. Dermal fibroblasts were isolated and cultured with various concentrations of ALA for 6 h. Intracellular protoporphyrin IX (PpIX) was measured by confocal microscopy. After 5 h of ALA treatment, cells were irradiated by a red laser (635 nm wavelength) at a power density of 10 mW/cm(2) with an energy density from 0.5 to 4 J/cm(2). Cell survival was measured by a CCK-8 Kit after 24 h. Cell death was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and flow cytometric analysis of annexin V. Results: Intracellular PpIX accumulation was observed in fibroblasts from HS patients and healthy donors after ALA treatment. After laser irradiation, viable cells were decreased among both cell types in a dose-dependent manner with energy density. In addition, apoptotic cell death was observed with low dose PDT, whereas necrotic cells were increased by high dose PDT. Conclusion: HS-derived fibroblasts efficiently accumulate PpIX after ALA treatment and can be eliminated via apoptosis by controlled laser irradiation.