Abstract
The decellularization of long segments of tubular tissues such as blood vessels may be improved by perfusing decellularization solution into their lumen. Particularly, transmural flow that may be introduced by the perfusion, if any, is beneficial to removing immunogenic cellular components in the vessel wall. When human umbilical arteries (HUAs) were perfused at a transmural pressure, however, very little transmural flow was observed. We hypothesized that a watertight lining at the abluminal surface of HUAs hampered the transmural flow and tested the hypothesis by subjecting the abluminal surface to enzyme digestion. Specifically, a highly viscous collagenase solution was applied onto the surface, thereby restricting the digestion to the surface. The localized digestion resulted in a water-permeable vessel without damaging the vessel wall. The presence of the abluminal lining and its successful removal were also supported by evidence from SEM, TEM, and mechanical testing. The collagenase-treated HUAs were decellularized with 1% sodium dodecyl sulfate (SDS) solution under either rotary agitation, simple perfusion, or pressurized perfusion. Regardless of decellularization conditions, the decellularization of HUAs was significantly enhanced after the abluminal lining removal. Particularly, complete removal of DNA was accomplished in 24 h by pressurized perfusion of the SDS solution. We conclude that the removal of the abluminal lining can improve the perfusion-assisted decellularization.
Highlights
The decellularization of long segments of tubular tissues such as blood vessels may be improved by perfusing decellularization solution into their lumen
For all the subsequent experiments involving the abluminal lining removal, human umbilical arteries (HUAs) were treated with the collagenase solution containing 50% sucrose for 60 min as the treatment resulted in acceptable vessel permeability with better consistency
We demonstrate the presence of an abluminal watertight lining in HUAs that renders the vessel water impermeable
Summary
The decellularization of long segments of tubular tissues such as blood vessels may be improved by perfusing decellularization solution into their lumen. Decellularization is a process to remove immunogenic cellular components from native tissues or organs, while minimizing the associated adverse effects on the chemical composition, biological activity, and mechanical properties of their extracellular matrix (ECM). It has been found that non-chemically cross-linked decellularized tissues, when implanted, are able to modulate host response toward a constructive tissue remodeling[15] Because of their advantageous mechanical properties, decellularized native blood vessels have been used as a scaffold to create tissue-engineered vascular grafts (TEVGs)[11,16,17]. The diffusion of cellular components out of a long vessel may be limited to the abluminal side of the vessel
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