Decellularization of Caprine Urinary Bladder Using Sapindus Mukorossi Fruit Pericarp Extract

Abstract

In the present study herbal detergent has been used for decellularization of caprine urinary bladder matrix to develop scaffolds for use in regenerative medicine. Urinary bladder was delaminated and decellularized by hypertonic solution and 5% aqueous extract of Sapindus mukorossi fruit pericarp, respectively. Complete delamination was observed at 9±1h interval. The delaminated urinary bladder tissues were completely decellularized at 72 h as evidenced by histology, DAPI staining, scanning electron microscopy and estimation of DNA content. The herbal decellularization system removed ~94% of tissue’s DNA, preserving ECM’s microarchitecture. The non-enzymatic and collagenase (20U/ml) digestion test proved that the glutaraldehyde treatment significantly improve the biostability of these scaffolds. The free protein and free amino acid group concentration of uncross-linked decellularized matrix was significantly (P<0.05) higher than cross-linked tissues at different cross-linking time intervals. The fixation index was highest in tissues cross-linked for 72h. The free hydroxyproline content in the cross-linked acellular scaffolds was nil and moisture contents was significantly lower (P<0.05). Crosslinked scaffolds did not show any band pattern in SDS-PAGE gel. It was concluded that soap nut pericarp extract is a viable option for decellularization of caprine urinary bladder and glutaraldehyde treatment enhance its biostability. To the best of our knowledge, this is the first report on decellularization of urinary bladder using herbal detergent.