Abstract:

Objective To evaluate the different concentrations of sodium dodecyl sulfonate（SDS）on efficiency of cell removal from porcine small intestinal submucosa（SIS）and optimize the best concentration to produce an acellular porcine SIS scaffold for use in developing a tissueengineered corneal epithelium. Methods Fresh porcine SIS were decellularized with SDS (concentration of 0.1%, 0.2%, 0.3%, and 0.5% for 15 minutes, 30 minutes, 1 hour, and 2 hours) and randomly divided into 4 groups (A, B, C, D, 20 pieces in each group ) and the decellularization process was observed at the same time.After decellularization，the scaffolds were examined through the HE, 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) staining and the genomic DNA contect analysis to observe the biological characteristics of SIS. Results A cellular SIS was shown ivory, translucent and had certain transmittance. HE and DAPI staining revealed that when treatment with the time of 30 minutes in group A and B SIS were decellularized completely and preserved the overall tissue histoarchitecture, the collagen fiber porosity was increased, the acellular rate was more than 90%. Conclusion The acellular matrix treatment effect of 0.1% and 0.2% SDS for a period of 30min is satisfactory. The immunogenicity of the SIS can be reduced, therefore, it is an ideal acellular concentration for SIS and provides a theoretical basis for the construction of tissue engineering corneal epithelium.

Key words: Tissue engineering, Scaffold, Decellularization, Small intestinal submucosa, HE staining, DAPI staining