[Construction and evaluation of the property of decellular porcine aortic valve].
OBJECTIVE
To construct decellular porcine aortic valve (PAV) and to observe the existence of porcine endogenous retrovirus (PERV) and valve scaffold structure before and after implantation.


METHODS
(1) Porcine aortic valve was obtained. The cellular components of PAV were completely removed by using detergent and nucleotidase solution combined with alteration of osmosis. (2) The decellular underwent HE staining and light microscopy and detection of its physical and chemical properties. (3) 20 pieces of decellular PAV were implanted into dogs. On e month later blood samples of the dogs were collected. PCR and RT-PCR were used to detect the PERV expression in 20 samples of pig's peripheral blood, 20 fresh PAVs, cultured pig kidney cells of the PK15 line (as positive control), decellular PAVs implanted into the dogs, and 10 samples of dogs' peripheral blood. (4) Small pieces of decellular PAVs were implanted into the subcutaneous tissues of 6 rabbits at the back, 6 pieces for one rabbit, and then extracted by the ends of the 4th, 6th, 8th and 10th week respectively after implantation to undergo HE staining and light microscopy.


RESULTS
(1) Almost all cellular components in the PAVs had been removed after decellularization; the soluble protein contents lost markedly [(0.238 +/- 0.038)% vs. (0.484 +/- 0.116)%]; the water content of the decellular tissues increased significantly [(92.16 +/- 1.48)% vs. (89.2 +/- 1.55)%]; however, the decellular PAVs still maintained their excellent fibrous scaffold structure, and their shrinkage temperature and tension at fracture were not significantly changed [(72.0 +/- 0.7) degrees C vs. (71.2 +/- 0.8) degrees C, and (448.7 +/- 18.65)g/mm2 vs. (540.7 +/- 19.46)g/mm2 respectively]. (2) Agarose gel electrophoresis of all fresh PAVs and porcine peripheral blood samples showed a 219 bp band, which was 90% to 97% homologous with PERV-C gene, and the sequence of which is published in Medline. No 219 bp amplified band was found in all decellular PAVs and the peripheral blood samples of the dogs implanted with decellular PAV one month after the implantation. (3) The PAVs implanted in rabbit body showed very slight tissue reaction. Neutrophil, lymphocyte and plasmacyte infiltration were seen 4 weeks after; such inflammatory cell infiltration decreased markedly and the peripheral portions of the decellular PAVs began to be absorbed by the end of the 6th week after implantation. The decellular PAVs were completely absorbed without fibrosis or scar formation in the implantation area by the end of the 10th week.


CONCLUSIONS
(1) The cellular components of PAV can be completely removed, the excellent fibrous scaffold structure and mechanical strength of aorta valve can be maintained, and the antigenicity is very weak. Subcutaneous implantation investigation shows that decellular PAV is an absorbable and degradable biological material. (2) There is PERV-C in PAV that can be removed after decellularization. PERV-C reaction is negative in the peripheral blood samples of the recipients implanted with decellular PAV.
Liu
Wei-yong
WY
Department of Thoracic and Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Jin
Zhen-xiao
ZX
Gu
Chun-hu
CH
Zhao
Dong-e
DE
Tan
Hong-mei
HM
D000818:Animals; D001021:Aortic Valve / Q000166:cytology; D001705:Bioprosthesis; D004285:Dogs; D020077:Endogenous Retroviruses / Q000302:isolation & purification; D006349:Heart Valve Diseases / Q000601:surgery; D006350:Heart Valve Prosthesis; D008567:Membranes, Artificial; D011474:Prosthesis Design; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*
D008567:Membranes, Artificial