Abstract
Background5-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.MethodsTumor 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.ResultsThe 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.Conclusions5-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.
Highlights
Myxofibrosarcoma is a soft tissue sarcoma known for a high rate of local recurrence [1]
Fluorescence persisted after 5ALA removal in the MUG-Myx1 cell pellets but not in the adipose-derived stromal (ADS) cell pellets, where fluorescence quickly diminished to baseline levels
photodynamic therapy (PDT) MUG-Myx1 and ADS cells were treated with 5-Aminolevulinic acid (5-ALA) at a concentration of 500 μg/mL when they had both reached 70% confluence
Summary
Myxofibrosarcoma is a soft tissue sarcoma known for a high rate of local recurrence [1]. It is one of the most common sarcomas in elderly patients, mainly affecting patients during the sixth to eighth decades of life [2]. Myxofibrosarcoma typically occurs in the extremities and is less common in the trunk, neck, and feet. Curative treatment for extremity soft tissue sarcomas requires resection with wide margins. Despite frequently being located superficial to the fascia, myxofibrosarcoma tends to spread along the fascial planes with associated large areas of peritumoral edema. Because it is difficult to identify the extent of microscopic fascial spread, surgical treatment may require multiple reexcisions to obtain microscopically negative margins. Preoperative magnetic resonance imaging can aid in surgical planning, but tumor size is often underestimated [6]
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