Research Assistant Hofstra University Valley Stream, New York, United States
Introduction: The recent strategy of decellularizing plants to create scaffolds for vascular, skeletal, bone and cardiac tissue engineering has displayed immune responses during implantation. The need to negate the host immune response to implanted plant-based scaffolds has inspired a method of treatment to break down the molecular components of a leaf that elicit this response while maintaining its original structure. This cutting-edge technique is inexpensive and produces a biomimetic material that is biocompatible. The purpose of this study is to prevent plant-based scaffolds from generating an automatic immune response in vitro. Here we present an innovative method to develop grafts that utilizes decellularized leaf samples and a surface treatment that generates novel applications for vascular implementation.
Materials and
Methods: Leatherleaf viburnum scaffolds were pretreated in an alkaline solution at elevated temperatures. These scaffolds were taken out of the solution every quarter hour at each respective temperature. Surface-treated scaffolds were decellularized using 2% SDS for 72 hours, followed by a 10% bleach and 0.1% Triton X-100 solution for 3-6 hours. 3D vascular grafts were produced by rolling the decellularized scaffolds around a 1.5 mm rod with a gelatin and glutaraldehyde mixture. Tensile testing was performed upon each graft. Immune response testing was completed by seeding white blood cells onto each scaffold at a concentration of 625,000 cells/graft. Grafts were placed within a bioreactor set at 37˚C overnight to regulate temperature. Hoestch and trypsin treatments were then applied to quantify the white cell viability after 24 hours.
Results, Conclusions, and Discussions: We present an innovative method for generating 3D scaffolds utilizing decellularized leaf samples and a surface treatment solution. The tensile stress of treated samples was comparable to non-treated samples (6.34±5.0 MPa). Hoestch and trypsin treatments displayed evidence of successful increased white cell viability compared to the untreated control, suggesting immune response negation of treated scaffolds (105,000± 80,000 cells/graft). These plant-derived scaffolds were able to withstand moderate tensile stress and maintain their original structure, compared to commonly used vascular graft materials. Temperature optimization is further needed as results exhibited extreme fluctuations due to temperature variability. To summarize, we developed plant-based 3D scaffolds for vascular repair that can withstand necessary mechanical stress and annul the automatic immune response. Future studies will include different methods of surface treatment, temperature regulation, and biodegradation. The development of appropriate 3D scaffolds is hitched upon the improvements of these endeavors.
