In Vivo Antitumor Efficacy Analysis of PDT: A Technique to Determine the Phototoxic Potential of a Photosensitizer in Tumor Bearing Mice

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. This video describes the in vivo anti-tumor efficacy analysis using photosensitizer containing nanoparticles in a tumor-bearing mouse, to determine the phototoxic potential of the photosensitizer.

Protocol

All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Synthesis of Hyaluronic Acid-Ceramide (HACE)

1. 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.
2. 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.
3. Dissolve the synthesized....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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