In vitro tissue engineering of a cardiac graft using a degradable scaffold with an extracellular matrix–like topography Osamu Ishii, MD, Michael Shin,

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Presentation transcript:

In vitro tissue engineering of a cardiac graft using a degradable scaffold with an extracellular matrix–like topography Osamu Ishii, MD, Michael Shin, PhD, Taijiro Sueda, MD, PhD, Joseph P. Vacanti, MD The Journal of Thoracic and Cardiovascular Surgery Volume 130, Issue 5, Pages 1358-1363 (November 2005) DOI: 10.1016/j.jtcvs.2005.05.048 Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 A, Gross view of single-layer scaffold. A thin, nonwoven fibrous mesh is suspended across a wire ring with a diameter of about 15 mm. The thickness of the mesh is about 10 μm. The wire ring acts as a passive load to condition cardiomyocytes during in vitro culture. B, Scanning electron microscopic micrograph of scaffold. The electrospun fibers have an average diameter of 250 nm. The pores are interconnected and are much larger than the fibers. The topography of the nonwoven mesh resembles that of an extracellular matrix and facilitates cell attachment. The Journal of Thoracic and Cardiovascular Surgery 2005 130, 1358-1363DOI: (10.1016/j.jtcvs.2005.05.048) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 A, Cross-sectional view of a single graft. Hematoxylin and eosin staining shows that cells have attached to the scaffold and proliferate well through the entire thickness of the scaffold. (Original magnification 400×.) B, Cross-sectional view of a 5-layer graft. Cardiomyocytes were first cultured on individual scaffolds to induce cell attachment. After 5 to 7 days, 5 scaffolds were gently placed on top of each other and maintained in culture. The 5-layer construct has the appearance of a single tissue segment, as shown here with hematoxylin and eosin staining. There is no evidence of physical gaps, and it appears the individual layers have fused well. (Original magnification 400×.) C, Cross-sectional view of a single graft. Masson trichrome staining reveals the presence of muscle and collagen throughout the entire specimen. (Original magnification 400×.) D, Cross-sectional view of a 5-layer graft. Masson trichrome staining reveals the presence of collagen. In addition, a stratified cardiac muscle–like structure can be seen. (Original magnification 400×.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 1358-1363DOI: (10.1016/j.jtcvs.2005.05.048) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 A, Surface view of a 5-layer graft visualized with hematoxylin and eosin staining. Cells are spread across the entire surface. The appearance of overlapping nuclei is attributed to cells adhering to pores in the scaffold below the surface. (Original magnification 200×.) B, Surface view of a 5-layer graft visualized with Masson trichrome staining. Cardiac muscle–like structures can be seen. (Original magnification 200×.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 1358-1363DOI: (10.1016/j.jtcvs.2005.05.048) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 A, Immunohistochemical staining for cardiac troponin I in a 5-layered graft. Cardiac troponin I–positive cells are seen throughout the entire cross-section of the graft. (Original magnification 400×.) B, Immunohistochemical staining for collagen type I in a 5-layered graft. Collagen is abundantly present throughout the entire specimen. (Original magnification 400×.) C, Immunohistochemical staining for connexin43 in a 5-layered graft. Diffuse gap junctions between the cells on 5-layered grafts can be seen. The nuclei have been counterstained with 4′,6′-diamidino-2-phenylindole hydrochloride. (Original magnification 400×.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 1358-1363DOI: (10.1016/j.jtcvs.2005.05.048) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 A, Scanning electron microscopic micrograph of a 5-layer graft. The surface is densely covered with cells after 2 weeks of culture. Scale bar = 20 μm. B, Scanning electron microscopic micrograph of a 5-layer graft. The cells have penetrated the entire cross-section of the specimen. Nondegraded fibers of the scaffold are also present. Scale bar = 10 μm. The Journal of Thoracic and Cardiovascular Surgery 2005 130, 1358-1363DOI: (10.1016/j.jtcvs.2005.05.048) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions