Validation and Optimization of Porcine Adipose Tissue Scaffolds

Tissue engineering is an alternative approach to address the scarcity of much needed donor tissues and organs.  Biodegradable porous structures (scaffolds) are used to guide the growth of cells into the necessary shape and function of the tissue to be replaced.  Although scaffolds created from animal tissues such as skin, bladder and intestine have seen clinical usage, they are limited by their thickness, area, and quality.  As an alternative, we examined generating scaffolds from porcine adipose tissue (pig fat), an inexpensive byproduct of the meat industry. To achieve decellularization and fat removal from the adipose tissue, the tissue was soaked in 100% ethanol for 30 minutes then placed in a 1:1 mixture of chloroform and methanol for 30, 60, 90, or 120 minutes. The first method proved to be very time consuming and involved harmful agents. A second decellularization method was formulated using a freeze-thaw process with samples in 30%, 50%, or 70% at -80 ^oC and room temperature. Samples were analyzed via H/E staining, compression testing for mechanical properties, and freeze drying for a qualitative measure of fat content. Four cycles of freezing-thawing were found to remove the cellular components of the tissue but were not sufficient to fully remove the fat content from the tissue.  Currently, additional freeze-thaw cycles and adjustments to the freezing solution are being explored to fully remove the fat content from the tissue.