Title:

Determining Suitable Leaf Species for Perfusion of Leaf-Derived Vascular Scaffolds (LeaVS)

In the United States, nearly 500,000 Americans are treated annually for burns, with 40,000 requiring hospitalization and advanced treatment, costing between $2-3 billion USD annually. The standard of care is a split-thickness skin autograft, but due to limited tissue availability, a bioengineered skin substitute may be necessary. Current bioengineered skin substitutes lack vasculature necessary to enhance graft take. Decellularized leaf scaffolds possess vasculature mimicking the structural architecture of human skin and have the potential to function as inexpensive bioengineered skin substitutes, as cellulose is biocompatible and elicits a minimal inflammatory response. This study aims to evaluate several leaf types for their decellularization ability, perfusable vascular network morphology, and xylem diameters. Selected leaves include baby kale (Brassica oleracea), leatherleaf viburnum (Viburnum rhytidophyllum), sweet basil (Ocimum basilicum), Chicago hardy fig (Ficus carica), sage (Salvia officinalis), and spinach (Spinacia oleracea) as a control. To decellularize the leaves, a series of rinses alternating between hexanes and deionized water were performed, followed by 5 days in 1% w/v sodium dodecyl sulfate solution, and 3-21 days in a 1% Tween 20, 3% bleach solution until colorless. Leaves were perfused with red dye through the petiole to examine vasculature and analyzed with Vessel Analysis (ImageJ plugin) to quantify vascular density. Petiole xylem Feret diameters were analyzed by embedding samples in paraffin, sectioning, and imaging via phase contrast microscopy using a custom ImageJ macro. Results indicate that Chicago hardy fig and sweet basil have vascular density closest to spinach, mean Feret xylem diameters greater than spinach, and potential as decellularized scaffolds for seeding endothelial cells through the petiole. Additionally, leatherleaf viburnum and sage may be useful in drug delivery to a graft due to the uniform perfusion pattern observed during the dye perfusion test.

Name

Comment*

Comments are viewable only by submitter

Captcha*