Near infrared fluorescent chlorophyll nanoscale liposomes for sentinel lymph node mapping

Lina Fan,^1,* Qiang Wu,^1,* Maoquan Chu<sup>1,2

1</sup>School of Life Science and Technology, ²The Institute for Advanced Materials and Nano Biomedicine Tongji University, Shanghai, People's Republic of China

^*These authors contributed equally to this work

Background: Sentinel lymph node (SLN) mapping using in vivo near infrared fluorescence imaging has attracted great attention during the past few years. Here we report on the early use of poorly water-soluble chlorophyll with near infrared fluorescence extracted from the leaf of Chimonanthus salicifolius, for mouse axillary SLN mapping.
Methods and results: To improve the water solubility and SLN targeting of the chlorophyll, we encapsulated the chlorophyll in nanoscale liposomes. The liposome-coated chlorophyll nanocomposites obtained were spherical in shape and had an average diameter of 21.7 ± 6.0 nm. The nanocomposites dispersed well in water, and in aqueous suspension they exhibited brighter near infrared fluorescence than chlorophyll alone. After incubation of the nanocomposites with normal liver cells (QSG-7701) and macrophage cells (Ana-1) for no more than 48 hours, there was no obvious reduction in cell viability. When the nanocomposites were injected intradermally into the paw of a mouse, the axillary SLN was found to be strongly fluorescent and was easily visualized in real time without a requirement for surgery. The intensity of the near infrared fluorescence emitted by the SLN was obviously brighter than that emitted by the SLN of another mouse that had been intradermally injected with chlorophyll alone.
Conclusion: Our data show that the liposome-coated chlorophyll nanocomposites could have great potential for clinical SLN mapping due to their lack of toxicity, bright near infrared fluorescence, and small diameter.

Keywords: chlorophyll, liposomes, nanocomposites, near infrared fluorescence, sentinel lymph node mapping

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