Abstract
Introduction A major obstacle in tissue engineering is to create a surgically implantable tissue
with long-term viability. Several promising techniques have focused on biological
vascularized matrices (BioVaM) with preserved vascular pedicles in the porcine model.
However, the handling of this model is time-consuming and expensive. Therefore, our
aim was to establish a BioVaM in the rat.
Materials and Methods Small bowel segments of Sprague-Dawley rats were isolated and perfused via cannulation
of the superior mesenteric artery and the portal vein. All cellular matrix components
were removed by sequential treatment with sodium dodecyl sulfate, sodium deoxycholate,
and DNase. Quality of decellularization was investigated by histology and potential
residual DNA by spectrophotometry. Primary endothelial cells (ECs) isolated from the
major vessels of Sprague-Dawley rats. Cells were labeled with fluorescent cell tracker
and injected into the vascular pedicles of the matrix. Attachment of ECs was assessed
using fluorescence microscopy of the whole mount.
Results Decellularized matrix demonstrated the absence of cellular components but conserved
matrix architecture as determined by immune fluorescent, pentachrome, and hematoxylin
and eosin stains. DNA content was reduced by more than 99%. ECs were characterized
by specific staining against endothelial nitric oxide synthase and von Willebrand
factor; when injected, ECs attached along the vessel walls including the capillaries
of the intestinal wall.
Conclusions Rat small bowel segments harvested with intact vascular pedicles and associated vascular
network can be successfully decellularized and re-endothelialized ex vivo. This model
is an inexpensive and easy to handle alternative and appears to be a promising approach
for establishing vascularized tissue constructs.
Keywords
tissue engineering - rat model - vascular pedicles - vascularized tissue - re-endothelialization
