Application of Tergitol in liver decellularization and preparation of bioink based on the obtained extracellular matrix

Paweł Mirosław^1*Katarzyna Kosowska¹Wiktoria Serafin¹Radosław Piwowar¹Agnieszka Romanik-Chruścielewska¹Kinga Westphal¹Michał Wszoła^1,2,3Marta Klak^1,2*

• ¹Foundation of Research and Science Development, Warsaw, Poland
• ²Polbionica Ltd., Warsaw, Poland
• ³Medispace Medical Centre, Warsaw, Poland

Biological materials derived from decellularized tissues are of great interest in the field of tissue engineering. Artificial organoids can be used in drug discovery or toxicological studies, potentially reducing the reliance on animal models. They also serve as valuable models for cancer and virology research. In the study, 13 slightly different detergent-enzymatic decellularization processes were performed. The end products of each process were evaluated for their DNA content, residual lipids, sulfated glycosaminoglycans (sGAG), collagen and detergent residues. To our knowledge, fFor the first time, Tergitol was used in liver decellularization. A method for detecting residual Tergitol content in the matrix was also developed. A significant reduction in DNA and fat content was achieved, as well as preservation of collagen. However, a marked loss of sGAGs was observed. The most promising variant involved gradient decellularization using Tergitol and ammonium hydroxide combined with pre-digestion by trypsin, which yielded the most favorable results and showed no cytotoxicity. A bioink was subsequently formulated from the decellularized matrix, which has potential applications in 3D bioprinting of artificial liver tissue. Tergitol appears to be a promising agent for liver decellularization, but further studies are necessary to optimize the method for preserving soluble components of the extracellular matrix.