[Decellularized human tissue engineering aortic valves conduit].
OBJECTIVE
To explore the suitable method to create a decellularized human tissue engineering homograft aortic valves conduit and to study its biologic, immol/Lunological, and biomechanical properties.


METHODS
Human homograft aortic valves conduits donated by healthy adult males undergoing brain death preserved in liquid nitrogen were treated with pH 8.0 hypotonic Tris buffer for 12 hours; then pH 8.0 isotonic buffer with 0.5% DOA, 1% DOA or 1% Triton for 12 hours-24 h; l and then pH 7.6 isotonic buffer with DNAase 200 micro g/ml, RNAase 20 micro g/ml for 2 hours. the histology, HLA-DR antigen, water content, thinkness, denaturation temperature, DNA content, Collagen Contents, stress-strain, destroying stress were examined.


RESULTS
In comparison with the standard cryopreserved human homograft aortic valves conduit, the valves, wall and muscle of the homograft aorta treated by 1% DOA for 24 hours were decellularized completely, and the 3-dimensional network structure of elastic fibers and collagenous fibers remained intact. Immol/Lunohistochemistry showed a remarkable decrease of expression of tissue genome DNA contents, a decrease by 91.14% in the valves, and by 91.53% in the wall, and a remarkable decrease of the expression of HLA-DR antigens. However, the water content in the decellular aortic wall was increased significantly (75.4 +/- 1.8 vs 82.0 +/- 0.7, P < 0.05). No significant differences were found in the denaturation temperature, stress-strain, destroying stress parameter, and tissue extension ratio in the decellular human homograft aortic valves conduits.


CONCLUSIONS
The decellularization method by isotonic buffer with 1% DOA-DNAase is effective. The decellularized human homograft aortic valves conduits are unaltered in biologic, biomechanical, and physical properties with lowered immol/Lunogenicity, It can be used as an ideal valve for patients or as a homograft stent for developing tissue engineering valve by host recellularization.
Cui
Bin
B
Fuwai Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100037, China.
Liu
Ying-long
YL
Xie
Ning
N
Yu
Cun-tao
CT
Song
Lai-feng
LF
Li
Jian-rong
JR
Wu
Song
S
D000328:Adult; D001021:Aortic Valve / Q000166:cytology; D001705:Bioprosthesis; D006350:Heart Valve Prosthesis; D006801:Humans; D008297:Male; D011474:Prosthesis Design; D023822:Tissue Engineering / Q000379:methods; D014184:Transplantation, Homologous