J Korean Med Sci.  2007 Apr;22(2):262-269. 10.3346/jkms.2007.22.2.262.

Modified Acellularization for Successful Vascular Xenotransplantation

• Jo WM
• Sohn YS
• Choi YH
• Kim HJ
• Cho HD

• KMID: 1713185
• DOI: http://doi.org/10.3346/jkms.2007.22.2.262

Abstract

The purpose of this study was to estimate the possibilities of an acellular matrix using a modified acellularization protocol, which circumvents immunological, microbiological, and physiological barriers. We treated porcine subclavian arteries with various reagents to construct acellular grafts. Afterwards, these grafts were interposed in a mongrel dogs' abdominal aorta. Six dogs underwent interposition with fresh porcine grafts (control group), and seven had interposed acellular grafts (acellular group). The control and acellular group dogs were sacrificed at 1, 3, 5 (n=2 in each group) and 12 months (n=1 in acellular group) after the operation. Histopathological examinations were then performed, to assess the degree to which re-endothelialization, inflammation, thrombus formation, and calcification occurred. The entire acellular group, but none of the control group, exhibited re-endothelialization. The degrees to which inflammation, thrombosis, and calcification occurred were found to be lower in the acellular group. We also discovered many smooth muscle cells in the medial layer of the xenograft that had been implanted in the dog sacrificed 12 months after the operation. These results suggest that the construction of xenografts using our modified acellularization protocol may offer acceptable outcomes as a vascular xenograft.

Keyword

Xenograft; Vascular Endothelial Cell; Graft Rejection

MeSH Terms

Transplantation, Heterologous/*methods
Tissue Engineering/*methods
Swine
Subclavian Artery/*cytology/*transplantation
Graft Survival/*physiology
Dogs
Cell-Free System/*transplantation
Animals

Figure

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  Fig. 1 Endothelial cells of cellular matrix. (A) Dog aorta (hematoxylin and eosin stain). (B) Pig graft (hematoxylin and eosin stain). (C) Immunohistochemical staining of dog aorta with rabbit anti-human Factor VIII antibody, which stained the endothelial cytoplasm positively (arrow). (D) Immunohistochemical staining of pig graft with rabbit anti-human Factor VIII antibody, which did not stain endothelial cytoplasm.

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  Fig. 2 Cellular matrix (Control, A, B) and acellular matrix (C, D); Cellular matrix (A, B) showed many cellular materials and stained nuclei, but acellular matrix (C, D) did not contain any cellular materials and showed collagen fibers only. (A) Cellular matrix (control group, hematoxylin and eosin stain, ×100). (B) Cellular matrix (control group , EM, ×10,000). (C) Acellular matrix (acellular group, hematoxylin and eosin stain, ×200). (D) Acellular matrix (acellular group, EM, ×8,000).

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  Fig. 3 Immunohistochemical stain of the recipient's endothelial cells with rabbit anti-human Factor VIII antibody. Control group graft did not reveal the re-endothelialization, but the entire acellular group showed re-endothelialization and repopulation from the recipient's cells. (A) Control group did not reveal re-endothelialization 3 months after operation. (B) Recipient's endothelial cells revealed ingrowth into the acellular graft (arrow) 3 months after operation. (C) Recipient's endothelial cells showed ingrowth in the acellular graft (arrow), and several cells were noted in the medial layer (empty arrow) 12 months after operation.

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  Fig. 4 Immunohistochemical stain of medial layer with smooth muscle actin (SMA). (A) Recipient's (dog) normal abdominal aorta. It showed a regular arrangement of the normal smooth muscle cells. (B) Donor's (pig) xenograft. The en-growing cells in the medial layer proved to be positive for SMA immunostaining but arranged irregularly, which had migrated from the recipient's aorta.

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