In Vitro Phototoxicity Assay: A PDT-based Method to Evaluate the Phototoxic Potential of a Photosensitizer in Lung Cancer Cells
Published: April 30, 2023
Abstract
Source: Chang, J. E., et al. Anticancer Efficacy of Photodynamic Therapy with Lung Cancer-Targeted Nanoparticles. J. Vis. Exp. (2016).
Photodynamic therapy (PDT) is a non-invasive and non-surgical method for lung cancer treatment. Photosensitizers selectively accumulate in tumor tissue and lead to tumor cell death in the presence of oxygen and the proper wavelength of light. In this protocol video, PDT-based method is used to evaluate the phototoxic potential of a potential photosensitizer in lung cancer cells.
Protocol
1. Synthesis of Hyaluronic Acid-Ceramide (HACE) Solubilize 12.21 mmol of hyaluronic acid (HA) oligomer and 9.77 mmol of tetra-n-butylammonium hydroxide (TBA) in 60 ml of double-distilled water (DDW). Stir for 30 min. To synthesize the DS-Y30 linker, dissolve 8.59 mmol of DS-Y30 ceramide and 9.45 mmol of triethylamine in 25 ml of tetrahydrofuran (THF). Mix with 8.59 mmol of 4-chloromethylbenzoyl chloride in THF. Stir for 6 hr at 60 °C. Dissolve the synthesized 8.10 mmol of HA-…
Disclosures
The authors have nothing to disclose.
Materials
| oligo hyaluronic acid | Bioland Co., Ltd. | ||
| DS-Y30 (ceramide 3B; mainly Noleoyl-phytosphingosine) | Doosan Biotech Co., Ltd. | ||
| adipic acid dihydrazide | Sigma Aldrich | A0638 | |
| N-(3-dimethylaminopropyl)-N′- ethylcarbodiimide | Sigma Aldrich | 39391 | |
| 4-(chloromethyl)benzoyl chloride | Sigma Aldrich | 270784 | |
| Paclitaxel | Taihua Corporations | ||
| syringe filter | Sartorius Stedim Biotech GmbH | 17762 | 15 mm, RC, PP, 0.45 µm |
| RPMI-1640 | Gibco Life Technologies, Inc. | 11875 | |
| Penicillin–streptomycin | Gibco Life Technologies, Inc. | 15070 | |
| Fetal bovine serum | Gibco Life Technologies, Inc. | 16140071 | |
| Celite (Filter agent) | Sigma Aldrich | 6858 | See step 1.4 |
Cite This Article
In Vitro Phototoxicity Assay: A PDT-based Method to Evaluate the Phototoxic Potential of a Photosensitizer in Lung Cancer Cells. J. Vis. Exp. (Pending Publication), e20248, doi: (2023).