Evaluation of in vivo models for studying calcification behavior of commercially available bovine pericardium.

BACKGROUND AND AIM OF THE STUDY: A common frame of reference is essential when attempting to determine if new treatments intended to reduce calcification of bioprostheses are superior to existing processes and products. The aim of this study was to examine calcification behavior for a commercially available pericardial bioprosthesis in subcutaneous and sheep valve models, and to evaluate the importance of appropriate control treatments in comparative studies with proposed new treatments. METHODS: Samples of bovine pericardium were placed subcutaneously under the dorsal skin of weanling rats and juvenile rabbits for 30-, 60- and 90-day intervals. Samples were either commercially available pericardial tissue or tissue processed with phosphate-buffered glutaraldehyde alone. Commercially available pericardial valves were also implanted in the mitral position in juvenile sheep, with elective sacrifice at 20 weeks. Retrieved samples underwent X-ray, histologic and elemental analysis. RESULTS: Commercial samples retrieved from the subcutaneous and sheep models showed similar, minimal calcification behavior on X-ray and histologic slides, whereas pericardium exposed to glutaraldehyde alone demonstrated rapid calcification. CONCLUSIONS: The 90-day subcutaneous rabbit model produced patterns of calcification similar to those in valves explanted from juvenile sheep after 20 weeks. A statistically significant decrease (p < 10(-8)) in calcification was demonstrated for clinical pericardium when compared with pericardium exposed to glutaraldehyde alone in the subcutaneous model. This suggests that subcutaneous models may be a cost-effective, time-efficient means of evaluating and comparing various tissue treatment methods. The rabbit methodology may provide a more accurate prediction of clinical performance, offering a greater degree of sensitivity. These studies also indicate that the commercially available process shows minimal calcification in the commonly used 30-day weanling rat subcutaneous model, contradicting other reported studies that may not accurately represent commercially available processes.

Evaluation of epoxy ether fixed bovine arterial grafts for mutagenic potential.

Two epoxy ether compounds [glycerol polyglycidyl ether (Denacol EX-313) and ethylene glycol diglycidyl ether (Denacol EX-810)] are under consideration as alternatives to glutaraldehyde for use in the processing of an arterial graft. The two are utilized as cross-linking and sterilant agents, respectively. Epoxy resins are multifunctional alkylating agents, and bifunctional alkylating epoxide solutions are known to be mutagenic. The correlation between mutagenic potential and carcinogenicity, in addition to evidence that diepoxides are carcinogenic in mice and rats, prompted the evaluation of the mutagenic potential of the epoxy treated, clinically rinsed graft. Ames and sister chromatid exchange (SCE) test procedures were used to assess mutagenic potential. Normal saline and distilled water were selected as the most physiologically representative and procedurally acceptable extraction mediums for the Ames and SCE tests, respectively. The results of the Ames and SCE tests in both the activated and non activated systems indicated that there were no statistically significant differences detected between various test article concentrates and the spontaneous mutation controls for both the activated and non activated systems. The epoxy treated graft was determined to be non mutagenic and demonstrated no dose related responses by these methods.

Evaluation of two epoxy ether compounds for biocompatible potential.

Bovine arterial tissue exposed to two epoxy ether compounds (Denacol EX-313 and Denacol EX-810) was evaluated for its biocompatible potential by in vitro and in vivo test procedures. The battery of test procedures included percent Inhibition of Cell Growth, Medium Eluate Method (MEM), Agar Overlay (AO), Blood Compatibility, Acute Mouse Systemic Injection, Rabbit Intracutaneous Irritation, Rabbit Subcutaneous Implantation, Guinea Pig Maximization, and Ames Tests. The epoxy exposed tissue was found to be noncytotoxic, nonmutagenic, and biocompatible by the test methods employed. In addition, the maximum concentrations of the unreacted Denacol EX-313 and EX-810 solutions found to demonstrate noncytotoxic reactions by the MEM and AO procedures were identified as 55 and 60 ppm for the MEM and 150 and 200 ppm for the AO procedure, respectively. These studies suggest that Denacol EX-313 and EX-810 are acceptable solutions for the processing of implantable tissue provided the epoxy residuals remain below those levels found to be cytotoxic.