A Novel Perfusion Setup for In Vitro Calcification Analysis of Cardiac Patch Materials

Background: Calcification as a main degenerative mechanism remains a major problem in the long-term use of (biologic) patch material in pediatric cardiac surgery. To be able to evaluate the susceptibility of different materials to mineralization processes in vitro, we developed a standardized perfusion setup.

Methods: A centrifugal pump with its control module, hoses, a PTFE conduit, a heater unit, pressure lines, and a box were used to establish the perfusion setup. A calcification solution was chosen for perfusion. Perfused patch material was analyzed histologically (Alizarin red S staining) and chemically (micro X-ray fluorescence).

Results: The pump perfuses a parallel-connected hose system. In one perfusion line, patch material is sutured in the PTFE conduit. In the other line, a rubber sheath grounds the patch. Perfusion takes place at 37°C with 2,000 rpm. This creates a 5 L/min flow, which is partitioned into the parallel hose system to mimic conditions in pediatric patients. Perfusion pressure is set at 60 mm Hg. The pilot experiment showed that clear luminal patch calcification could be detected after 60 days of continuous perfusion.

Conclusion: Our self-made perfusion setup allows testing of different patches on their susceptibility to mineralization under standardized conditions. With our system, comparative analyses of different (commercially available) patches can be easily performed under in-vitro conditions in the future.

 

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No conflict of interest has been declared by the author(s).

Publication History

Article published online:
11 February 2025