Development of Ficus Benjamina Leaves Derived Cellulose Scaffolds for Tissue Engineering Applications

Abstract

The main aim of this study was to prepare Ficus benjamina leaves scaffolds for tissue engineering applications. Fresh leaves were decutinized with mixed solution of n-hexanes and diethyl ether in ratio of 9:1 and decellularized using 2.5% SDS (P1 and P2) and 5% SDS (P3 and P4) followed by incubation in 0.5% (P1, P3) and 1% (P2, P4) TnBP solution and finaly soaking in 4% NaOCl for 12 to 36h. The control and decellularized leaves scaffolds samples were characterized on the basis of macroscopic observations, histological/microscopic observations, DAPI staining, DNA quantification, density and porosity, swelling property, water vapor transmission rate, FTIR spectroscopic examination, and biocompatibility assessment. Cuticular wax was completely removed by n-hexane and diethyl ether treatment and the treatment leaves were completely decellularized at 12 and 24h interval after bleaching as revealed by H&E, DAPI stained micrographs and SEM ultrastructure. DNA (ng/µl) quantity, density and of the decellularized Ficus benjamina leaves tissue also reduced significantly (P<0.05). However, porosity, swelling percent and water vapor transmission rate of decellularized leaf scaffolds increased significantly (P<0.05). Decellularized leaf scaffolds were found hemocompatible. FTIR analysis of both control and decellularized Ficus benjamina leaf samples showed well defined similar peaks in spectra. The subcutaneously implanted decellularized leaf scaffolds were infiltrated with host cells and found biocompatible as showed by histopathological, DAPI and SEM images. The results demonstrated that the use of SDS and TnBP by immersion technique is a suitable procedure for decellularization of Ficus benjamina plants leaves to produce biocompatible cellulose scaffolds.