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5-Aminolevulinic acid tumor paint and photodynamic therapy for myxofibrosarcoma: an in vitro study
Journal of Orthopaedic Surgery and Research ( IF 2.8 ) Pub Date : 2020-03-05 , DOI: 10.1186/s13018-020-01606-9
Shachar Kenan , Haixiang Liang , Howard J. Goodman , Andrew J. Jacobs , Amanda Chan , Daniel A. Grande , Adam S. Levin

5-Aminolevulinic acid (5-ALA), a fluorescent contrast agent, has been used for tumor paint and photodynamic therapy (PDT) for various tumors, but its use with soft tissue sarcomas is not well documented. Myxofibrosarcoma, a subtype of soft tissue sarcoma with a high local recurrence rate, may benefit from similar types of treatment. The purpose of this study was to analyze the effects of 5-ALA tumor paint and PDT on a myxofibrosarcoma cell line. Tumor paint was assessed by exposing micromass pellets of human adipose-derived stromal (ADS) cells or myxofibrosarcoma (MUG-Myx1) cells to 5-ALA. Cell pellets were then visualized using a microscope at established excitation and emission wavelengths. Corrected total cell fluorescence was calculated per accepted protocols. Photodynamic therapy was similarly assessed by exposing ADS and MUG-Myx1 cells to 5-ALA, with subsequent analysis via flow cytometry and real-time confocal microscopy. The use of 5-ALA tumor paint led to a selective fluorescence in MUG-Myx1 cells. Findings were confirmed by flow cytometry. Interestingly, flow cytometry results showed progressive selective cell death with increasing 5-ALA exposure as a result of the PDT effect. PDT was further confirmed using confocal microscopy, which revealed progressive cellular bubble formation consistent with advancing stages of cell death—a finding that was not seen in control ADS cells. 5-ALA tumor paint and PDT were successfully used on a human myxofibrosarcoma cell line (MUG-Myx1). Results from this study showed both selective fluorescent tagging and selective cytotoxicity of 5-ALA toward malignant myxofibrosarcoma cells, while sparing benign adipose control cells. This finding was further confirmed in a dramatic time-lapse video, visually confirming active, targeted cell death. 5-ALA’s two-pronged application of selective tumor identification and cytotoxicity may transform surgical and medical approaches for treating soft tissue sarcomas.

中文翻译:

5-氨基乙酰丙酸肿瘤涂料和光动力疗法治疗粘液原纤维肉瘤:体外研究

5-氨基乙酰丙酸(5-ALA),一种荧光造影剂,已被用于多种疾病的肿瘤涂料和光动力疗法(PDT),但其与软组织肉瘤的使用尚无充分文献记载。粘膜纤维肉瘤是一种软组织肉瘤的亚型,具有较高的局部复发率,可以从相似的治疗方法中受益。这项研究的目的是分析5-ALA肿瘤涂料和PDT对粘膜原纤维肉瘤细胞系的影响。通过将人脂肪来源的基质细胞(ADS)或粘原纤维肉瘤(MUG-Myx1)细胞的微团沉淀暴露于5-ALA来评估肿瘤涂料。然后使用显微镜在确定的激发和发射波长下观察细胞沉淀。根据接受的方案计算校正的总细胞荧光。通过将ADS和MUG-Myx1细胞暴露于5-ALA进行类似的光动力学疗法评估,随后通过流式细胞术和实时共聚焦显微镜进行分析。5-ALA肿瘤涂料的使用导致MUG-Myx1细胞产生选择性荧光。通过流式细胞术证实了发现。有趣的是,流式细胞术结果显示,PDT效应导致5-ALA暴露增加导致选择性细胞死亡。使用共聚焦显微镜进一步证实了PDT,该技术揭示了与细胞死亡的进展阶段相一致的进行性细胞气泡形成-在对照ADS细胞中未发现这一发现。5-ALA肿瘤涂料和PDT已成功用于人类粘膜纤维肉瘤细胞系(MUG-Myx1)。这项研究的结果表明5-ALA对恶性粘液原纤维肉瘤细胞的选择性荧光标记和选择性细胞毒性,同时保留了良性脂肪对照细胞。这一发现在一个引人注目的延时录像中得到了进一步证实,该录像在视觉上证实了活跃的靶向细胞死亡。5-ALA对选择性肿瘤识别和细胞毒性的双重应用可能会改变用于治疗软组织肉瘤的手术和医学方法。
更新日期:2020-03-06
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