This study was carried out to estimate a good decellularization protocol for pericardium and jugular vein of cattle and buffalo with preservation of the extracellular matrix.
Pericardium and jugular vein were decellularized chemically using Triton X-100 (TX)
plus sodium deoxycholate (SD), physically by freeze–thaw cycles +TX + SD and enzymatic protocol used TX+SD+ DNase +RNase. Untreated pericardium and jugular
vein were used as control. The histological analysis was performed to evaluate the
efficiency of decellularization and extracellular matrix preservation. But The Immunoreaction was evaluated by subcutaneous implantation in rats. It was found that no
cells or cell fragments were retained, and there was no apparent tissue disruption in
the decellularized tissues, except chemical group showed that the layers of pericardium were dispersed, and layers of jugular vein revealed that the elastic fibers were dispersed, degraded and disorganized. The content of collagen and elastic fibers were
affected after decellularization, in physical group showed mild detached collagen fibrils and mild fragmented elastic fibers, but enzymatic group makes
sever detached collagen fibrils and sever disrupted elastic fibers. The implanted tissue
showed mild-moderate inflammatory reaction and calcification in physical group, but
scanty to mild inflammatory reaction and calcification in enzymatic group. These results suggested that the physical protocol showed optimal decellularization results
with better extracellular matrix preservation than other methods and the enzymatic
method is good in reduction the immunoreaction (mild to scanty inflammatory cells
infiltration). |
