Development of human umbilical cord based scaffold for tissue engineering application

Abstract

Bioactive scaffolds composed of decellularized Extracellular Matrix (ECM) are attractive for Tissue engineering (TE) applications as such scaffolds provide natural biologic signals which are essential for the regeneration of tissues and exhibit excellent cell attachment and proliferation ability. This study compares different methods to optimise the decellularization of human umbilical cord (UC) tissues and subjecting these tissues to solubilization and film formation. The decellularization process consisted of a single freeze thaw cycle where the stored tissues at -20°C are thawed at room temperature followed by dissection of UC in appropriate sizes. Trypsin/EDTA and Peracetic acid was not successful in decellularzing UC tissues. Decellularization using Sodium dodecyl sulfate (SDS) and Triton-X 100 revealed that both 1% SDS and 0.1% SDS was successful in removing the cells from the small UC tissues (∼0.5cm). When the UC tissues were large (∼1 cm) only 1% SDS was able to completely decellularize cells. 1% SDS decelluarized UC tissues were ground in cryomill and solubilized in pepsin for 48 hours. UC tissues washed with several phosphate buffered saline (PBS) washes were not able to solubilize in porcine pepsin. Hence 1% SDS protocol was optimised by washing the decellularized tissues with 75% ethanol and was solubilized in pepsin. The film developed from the decellularized UC tissues support amniotic epithelial cell (AME) attachment and can be suitable for TE applications.