Novel Method of Sterilization of Porcine Liver Bio-Scaffold Using the Electrolyzed Water

Abstract

Introduction: Tissue-engineered liver bio-scaffolds are in need to be sterilized to meet the clinical application requirements. Although there are many techniques used for sterilization, each one has advantages and disadvantages which may adversely affect the ECM properties. Slightly acidic electrolyzed water (SlAEW) which contains free chlorine, is considered to be effective for disinfection. SlAEW is generated by the electrolysis of a solution containing low NaCl concentration. It has a strong bactericidal activity and disinfection potential against viruses including hepatitis B and C viruses. The aim of the study to develop a new technique which enables the researchers to sterilize not only the liver scaffold but also all kinds of scaffolds with minimal effects on the ECM properties. Methods: Fresh porcine livers were sliced and decellularized using 0.1%SDS. Decellularization was evaluated by histological and biochemical (DNA, glycosaminoglycan and collagen content) analyses. Sterilization was done by the slightly acidic electrolyzed water for different periods (20min,1hour & 2hours) for optimization the sufficient time for sterilization. Analysis of the sterilization efficiency was done by microbiological tests. The morphological and biochemical changes were analyzed after the sterilization. We also evaluated the effect of sterilization on the host cell attachment, proliferation and short term viability using in vitro bioactivity tests. Results: The use of SlAEW resulted in generation of a sterilized scaffold with minimal effect on the ECM components and structure. The in vitro bioactivity test revealed that scaffold treated by SlAEW can support the attachment, viability and proliferation of cells. Conclusion: We conclude that the biologic porcine liver scaffold can be sterilized with the electrolyzed water and still retain its bioactivity. This work was also supported by a grant from the Next-Generation BioGreen 21 program (No.P J008030,J008032), Rural Development Administration, Republic of Korea.