Poster AA17 - Optimization of Plant Decellularization for Vascular Graft Development

Introduction: In recent years the use of decellularized plant tissue as a biomaterial has become more prevalent due to the remarkable similarities between plant and animal tissues. Decellularized plants have many advantages such as their availability, low cost, and eco-friendly nature of their production. Since the use of plant-derived materials for tissue engineering is so new, it is crucial to begin standardizing these products with the intention of clinical applications in the future. The goal of this study is to evaluate the differences in extracellular matrix structure between plant tissue decellularization techniques to determine the most appropriate method for the development of 3D vascular grafts.

Materials and

Methods: Leatherleaf viburnum was cut to a standard dimension and decellularized using various techniques including the following: Condition A: trypsin, tergitol and EGTA; Condition B: trypsin, tergitol; Condition C: SDS, tergitol; and and condition E: SDS, 6 hours bleach, triton X-100 (clearing solution). The effect of the duration of treatment in the clearing solution was also evaluated on SDS-decellularized leatherleaf for up to 3 days. Samples were all decellularized using condition E. Samples were collected from four different locations on our campus, and were all decellularized using condition E, to determine if the leaves varied between locations. H&E and safranin staining was performed on each sample. Samples were imaged and image analysis was performed using ImageJ to evaluate the diameters of fibers within the cross-section of the plant. Additionally, the OrientationJ plugin was used to evaluate fiber orientation.

Results, Conclusions, and Discussions: As the use of decellularized plant tissue as a biomaterial becomes more frequent, the standardization of development of plant-based scaffolds and tissues has become critical to ensuring safe and effective manufacturing for future research and medical advancements. A one-way ANOVA yielded significant variation in fiber diameter between conditions, p< 0.05. A post hoc Tukey test showed that fiber diameter of condition B was significantly higher (by 23.42%, p< 0.05) for decellularized leatherleaf compared to conditions A and D. The structural difference between ECM derived from these decellularization protocols is important because it suggests potential functional differences between graft scaffolds, such as tissue mechanics and the ability to promote cell infiltration. These properties are crucial to the success of vascular graft materials after implantation. The study of plant decellularization protocols and the resulting ECM structures will help enable scaffold optimization for graft development.

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