Abstract:

BACKGROUND: Preparation of extracellular matrix (ECM) aims to remove the cellular components in tissues, there is no uniform standard for the current acellular methods, but the prepared ECM should be proved to be without residual cells. 
OBJECTIVE: To prepare human umbilical vein ECM by using hypotonic and hypertonic solutions, detergent and proteinase in a multistep process, mixed solution of Triton X-100 and ammonia water, and to explore the ideal method of preparing human umbilical vein ECM. 
METHODS: ECM of human umbilical veins was prepared by use of hypotonic and hypertonic solutions, detergent and proteinase in a multistep process and solution of Triton X-100 and NH₃·H₂O. The physical properties and hematoxylin-eosin staining of human umbilical vein before and after acellular matrix were observed, the amount of residual cell debris in 50 images at 400 magnification was measured.
RESULTS AND CONCLUSION: The acellular umbilical veins exhibited translucent porcelain-white jelly-like tubular structure. The cell components of the human umbilical veins resulting from hypotonic and hypertonic solutions, detergent and proteinase were almost removed, ECM maintained an integrity, and fibrous structures were preserved. ECM resulting from solution of Triton-X100 and NH₃·H₂O had different quantities of remaining cellular elements. There were significant differences between the two groups (P < 0.05). Hypotonic and hypertonic solutions, detergent and proteinase are ideal methods to prepare ECM from human umbilical veins.