Decellularization of a Fasciocutaneous Flap for Use as a Perfusable Scaffold

The aim of the current study was to determine whether a rat fasciocutaneous flap could be decellularized using detergent perfusion and/or agitation methods while preserving the integrity of the extracellular matrix and circulatory networks.

Superficial inferior epigastric arterial flaps of 50 rats were randomly divided into the following 5 groups: (1) normal; (2) agitation in sodium dodecyl sulfate (SDS) for 72 hours (72h-AG); (3) perfusion and agitation with SDS for 12 hours (12h-PE-AG); (4) perfusion and agitation with SDS for 24 hours (24h-PE-AG); and (5) perfusion and agitation with SDS for 72 hours (72h-PE-AG). These flaps were evaluated by gross morphology, histology, integrity of the microcirculatory networks, and DNA quantification.

The DNA content of the normal flap was 1.53 μg/mg. The decellularized flaps had significantly reduced DNA contents: 72h-AG (0.55 μg/mg), 12h-PE-AG (0.52 μg/mg), 24h-PE-AG (0.23 μg/mg), and 72h-PE-AG (0.17 μg/mg). The DNA contents in both the 24h-PE-AG and 72h-PE-AG groups were significantly less than that of 72h-AG and 12h-PE-AG groups. These findings were confirmed by histology and gross morphology. The integrity of the extracellular matrix and vascular system was preserved as measured by collagen and elastin stains in the 4 decellularized groups. Despite the histological appearance of vessel integrity, none of the flaps maintained physiologic vascular integrity by closed-loop circulation.

A combination of perfusion and agitation for 24 hours or longer effectively decellularized the fasciocutaneous portion of composite tissue flaps and removed DNA content from the flap in our rat model with well-preserved vascular structure. This combined technique was superior to agitation alone. However, closed-loop circulation could not be preserved after decellularization with perfusion and/or agitation methods.