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(Circulation. 2009;119:1653-1660.)
© 2009 American Heart Association, Inc.

Valvular Heart Disease

Detoxification and Endothelialization of Glutaraldehyde-Fixed Bovine Pericardium With Titanium Coating

A New Technology for Cardiovascular Tissue Engineering

Norbert W. Guldner, MD; Inka Jasmund, MS; Hanngörg Zimmermann, MS; Markus Heinlein, MS; Britta Girndt, MS; Veronika Meier, MD; Florian Flüß, MD; Daniel Rohde, MS; Andreas Gebert, MD; Hans-Hinrich Sievers, MD

From Klinik für Herzchirurgie (N.G., I.J., V.M., F.F., H.-H.S.), Universitätsklinikum Schleswig-Holstein, Lübeck, Germany; Gesellschaft für Elektrometallurgie Medizintechnik GmbH (H.Z., M.H., B.G.), Höfener Straße, Nürnberg, Germany; University of Applied Sciences (D.R.), Lübeck, Germany; and Institut für Anatomie (A.G.), Universität zu Lübeck, Lübeck, Germany.

Correspondence to Prof Dr Norbert W. Guldner, Klinik für Herzchirurgie Universitätsklinikum Schleswig-Holstein, Campus Lübeck, Ratzeburger Allee 160, D-233538 Lübeck, Germany. E-mail guldner{at}uni-luebeck.de

Received September 24, 2008; accepted January 8, 2009.

Background— Endothelial cell seeding of glutardialdehyde-fixed biological heart valves is hypothesized to improve biocompatibility and durability; however, the toxicity of glutardialdehyde prevents its use as a biological coating. Therefore, different detoxification strategies are applied, including surface coating with titanium, before in vitro endothelialization of glutaraldehyde-fixed bovine pericardium as the base material for prosthetic heart valves.

Methods and Results— Bovine pericardium was fixed with 0.25% glutardialdehyde. Detoxification was performed with citric acid, aldehyde dehydrogenase, and plasma deposition with titanium at low temperatures of 30°C to 35°C. Toxic glutaraldehyde ligands were quantified photometrically, and the vitality of seeded cells was tested to validate detoxification methods. Detoxification agents and titanium coating were applied before seeding with human endothelial cells. Endothelial cells were visualized by electron microscopic surface scanning. To evaluate cell adhesion, shear stress was applied by a flow of 5 L/min over 24 hours. Compared with untreated glutaraldehyde-fixed samples, treatment with the different agents reduced free aldehyde groups gradually (citric acid 5% < citric acid 10% < titanium < aldehyde dehydrogenase). A combination of citric acid 10%, aldehyde dehydrogenase, and titanium coating resulted in a reduction of free aldehyde ligands to 17.3±4.6% (P0.05) and demonstrated a vitality of seeded cells of 94±6.7% (P0.05). This procedure yielded a completely confluent layer of regular human endothelial cells (n=5). After application of shear stress for 24 hours on these endothelial layers, cell vitality was 81%.

Conclusions— Titanium coating combined with chemical procedures yielded significant detoxification and complete endothelialization of conventional glutaraldehyde-fixed pericardium. This new technique might improve glutardialdehyde-fixed cardiovascular bioimplants for better biocompatibility and longer durability.

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Clinical Summaries
Circulation 2009 119: 1553-1555. [Extract] [Full Text]