Quantification of Lymphangiogenesis in the Murine Lymphedema Tail Model Using Intravital Microscopy

Area of Science:

• Biomedical Engineering
• Vascular Biology
• Regenerative Medicine

Background:

• Lymphedema, a chronic swelling from lymphatic dysfunction, affects millions with no cure.
• Assessing lymphatic growth is crucial for developing new lymphedema therapeutics.
• Intravital microscopy (IVM) offers powerful in vivo imaging for biological processes.

Purpose of the Study:

• To evaluate the applicability of IVM for visualizing and quantifying lymphatic vessels.
• To assess lymphangiogenesis in a live murine tail lymphedema model using IVM.
• To investigate the efficacy of gene-based therapy delivered via tissue nanotransfection (TNT).

Main Methods:

• A murine tail lymphedema model was established.
• Tissue nanotransfection (TNT) delivered genetic cargo (pCMV6-Prox1 or pCMV6 control) to the surgical site.
• Confocal/multiphoton microscopy analyzed FITC-dextran stained lymphatic vessels and branch points.

Main Results:

• The TNT-Prox1 group showed reduced lymphedema and increased lymphatic distribution compared to controls.
• Over threefold more lymphatic branching points were observed in the TNT-Prox1 group.
• IVM successfully visualized and quantified lymphatic changes in vivo.

Conclusions:

• IVM is a novel and effective tool for studying lymphatic vessels in live lymphedema models.
• IVM enables functional assessment and visualization of lymphangiogenesis after gene therapy.
• This approach aids in evaluating novel therapeutics for lymphedema.