Release of Glutaraldehyde From an Albumin-Glutaraldehyde Tissue Adhesive Causes Significant In Vitro and In Vivo Toxicity Walter Fürst, MS, Asmita Banerjee

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Release of Glutaraldehyde From an Albumin-Glutaraldehyde Tissue Adhesive Causes Significant In Vitro and In Vivo Toxicity Walter Fürst, MS, Asmita Banerjee The Annals of Thoracic Surgery Volume 79, Issue 5, Pages 1522-1528 (May 2005) DOI: 10.1016/j.athoracsur.2004.11.054 Copyright © 2005 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Release of glutaraldehyde from BioGlue (1 mL) applied into four consecutive plates and overlaid with 5 mL of saline solution (mean ± minimum, maximum; n = 2 [two separate cartridges]). Plate 1 contained 1 mL of the material that is expelled when the cartridge is primed, and plates 2 to 4 contained BioGlue as it would be applied to tissues. Both the priming material that shall be discarded as well as the “save” material meant to be applied to the tissue released similar amounts of glutaraldehyde. The Annals of Thoracic Surgery 2005 79, 1522-1528DOI: (10.1016/j.athoracsur.2004.11.054) Copyright © 2005 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Carbolfuchsin staining of MRC5 cells incubated 100 minutes in cell culture medium supplemented with different concentrations of glutaraldehyde: (a) control; (b) 12 μg/mL; (c) 111 μg/mL; and (d) 333 μg/mL. The Annals of Thoracic Surgery 2005 79, 1522-1528DOI: (10.1016/j.athoracsur.2004.11.054) Copyright © 2005 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Cytotoxicity of supernatants obtained from BioGlue overlaid with saline solution. As assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide test, addition of these supernatants to the culture medium for 100 minutes nearly abolished viability of C2C12 mouse myoblast cells (mean ± standard deviation, n = 6). Two separate experiments with different cartridges of BioGlue were performed (Exp.1 and Exp.2). (BioGlueExp.1 and Exp.2 = culture medium supplemented with supernatants (10% vol/vol) of polymerized BioGlue; Medium = untreated cells; OD540 = optical density at 540 nm; saline = culture medium supplemented with saline [10% vol/vol].) The Annals of Thoracic Surgery 2005 79, 1522-1528DOI: (10.1016/j.athoracsur.2004.11.054) Copyright © 2005 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Effects of BioGlue and fibrin sealant (TISSEEL VH) on tissue of a rabbit partial lung resection model 2 days after application. (a) BioGlue (BG) evoked alveolar edema formation (X), while with fibrin sealant, alveoli (A) remained normally open (b). (c) Sham-operated animals. (Hematoxylin and eosin.) The Annals of Thoracic Surgery 2005 79, 1522-1528DOI: (10.1016/j.athoracsur.2004.11.054) Copyright © 2005 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Effect of BioGlue (BG) on the tissue of a rabbit liver after abrasion. Two days after application, areas of necrosis (a; circled area) with distinct boundaries (b; arrows) could be observed. (c) In contrast, fibrin sealant (FS) did not affect the liver tissue (L). Separation of fibrin sealant from tissue was caused by treatment for histologic evaluation. In vivo hemostasis was achieved, and no rebleeding was detected. (Hematoxylin and eosin.) The Annals of Thoracic Surgery 2005 79, 1522-1528DOI: (10.1016/j.athoracsur.2004.11.054) Copyright © 2005 The Society of Thoracic Surgeons Terms and Conditions

Fig 6 (a) Effect of BioGlue (BG) on aortic tissue. (b) Further magnification revealed no obvious reaction of aortic tissue. (A = aorta.) (Hematoxylin and eosin.) The Annals of Thoracic Surgery 2005 79, 1522-1528DOI: (10.1016/j.athoracsur.2004.11.054) Copyright © 2005 The Society of Thoracic Surgeons Terms and Conditions