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J Thorac Cardiovasc Surg 1997;114:421-427
© 1997 Mosby, Inc.

SURGERY FOR ACQUIRED HEART DISEASE

EFFECT OF CRYOPRESERVATION AND HISTOCOMPATIBILITY ON TYPE I PROCOLLAGEN GENE EXPRESSION IN AORTIC VALVE GRAFTS

Supported by grants 94-WA-532 and 95-WA-507R from the Washington Affiliate of the American Heart Association and by a grant from the Research Endowment of Children's Hospital and Medical Center.

Received for publication Dec. 26, 1996; accepted for publication April 28, 1997. Address for reprints: Flavian M. Lupinetti, MD, Division of Cardiac Surgery, Children's Hospital and Medical Center, 4800 Sand Point Way N.E., Seattle, WA 98105.

Abstract

Background: Allograft valves are excellent substitutes for diseased or absent valves but undergo primary tissue degeneration. Fibroblast viability may determine resistance to valve deterioration. This study evaluated gene expression for procollagen by valve grafts and studied the effects of cryopreservation and histocompatibility on this property. Methods and results: Fresh and cryopreserved rat aortic valves were implanted heterotopically into syngeneic or allogeneic recipients. Nonviable, cryothermally injured valves were used as negative controls. The grafts and native aortic roots were excised 3 days after implantation. Northern hybridization with a human procollagen ₁ (I) complementary DNA probe was used to assess the expression of type I procollagen mRNA. The content of procollagen mRNA relative to 18S ribosomal RNA was evaluated by means of scanning densitometry. In situ hybridization was used to locate the areas of procollagen mRNA expression in the grafts. Both fresh and cryopreserved grafts exhibited greater expression than the native valve. This increase in expression was observed in both syngeneic and allogeneic grafts, but not in the negative control group. In situ hybridization showed a strong signal for procollagen in the aortic wall and a weak signal in the leaflet and myocardium in the viable grafts and in native tissues. Conclusions: Regardless of preservation or allogenicity, fibroblast viability in aortic valve grafts persists after implantation. Increased gene expression for procollagen suggests a capacity for repair and regeneration.

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