208 FABRICATION OF A 3-D SCAFFOLD FOR SKELETAL MUSCLE TISSUE ENGINEERING

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2012208 FABRICATION OF A 3-D SCAFFOLD FOR SKELETAL MUSCLE TISSUE ENGINEERING Gjanje Smith, Jonathan McMichael, Heather-Marie Wilson, Robert Vernon, Thomas Kean, Robert Welikson, Kathleen Kobashi, James Dennis, and Margaret Allen Gjanje SmithGjanje Smith Seattle, WA More articles by this author , Jonathan McMichaelJonathan McMichael Seattle, WA More articles by this author , Heather-Marie WilsonHeather-Marie Wilson Seattle, WA More articles by this author , Robert VernonRobert Vernon Seattle, WA More articles by this author , Thomas KeanThomas Kean Seattle, WA More articles by this author , Robert WeliksonRobert Welikson Seattle, WA More articles by this author , Kathleen KobashiKathleen Kobashi Seattle, WA More articles by this author , James DennisJames Dennis Seattle, WA More articles by this author , and Margaret AllenMargaret Allen Seattle, WA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.261AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES A 3-D construct seeded with autologous cells on a biodegradable scaffold could serve as tissue replacement in pelvic floor reconstruction. Our goals are to design a scaffold that incorporates suture suitable for implantation and optimizes the conditions for a seeded implant in a 3-D construct by testing various formulations of collagen and hyaluronan. METHODS The C2C12 mouse myoblast cell line was used. Scaffold matrix components included collagen and hyaluronan (Hystem- HP®, Glycosan Biosystems). Constructs were seeded at 3, 4.5, and 6 ×105 cells/ml and polymerized in a silicone mold. 1:1,10:1 and 20:1 ratios of collagen:hyaluronan (C:H) were tested. Three scaffold designs were tested: constructs with a suture running through the middle, with Nitex mesh at the ends only, and with suture and mesh just inside the edges of the matrix. After 21 days, constructs were tested for passive stiffness. The constructs were then stained for dead cells, cell nuclei and filamentous actin, and confocal imaging was performed. RESULTS All construct conditions resulted in elongated, differentiated muscle cells with filamentous actin that appeared fused and multinucleated. C:H at 1:1 produced a more cellular construct with a greater proportion of elongated cells and fewer dead cells at 3 weeks than the 20:1 and 10:1 (C:H) samples (Figure 1). Constructs seeded with 6 ×105 cells/ml and C:H of 1:1 were 2x stiffer than those seeded at lower densities. The suture-only design (tension-free) resulted in random cell aggregation. Mesh alone also resulted in cell aggregation due to lack of integration of cells into the mesh. Combining suture and mesh allowed cells to integrate into the mesh and promoted rearrangement of the cells and matrix with development of uniaxial tension between suture ends which promoted cell elongation and alignment. CONCLUSIONS Greater cell viability was observed in constructs with an equal ratio of C:H. Combining suture with mesh located inside the matrix promoted the development of uniaxial tension that allowed the cells to align in the direction of tension. A higher seeding density resulted in a stiffer construct. Further testing is needed to determine the viability and functionality of constructs in vivo. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e87 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Gjanje Smith Seattle, WA More articles by this author Jonathan McMichael Seattle, WA More articles by this author Heather-Marie Wilson Seattle, WA More articles by this author Robert Vernon Seattle, WA More articles by this author Thomas Kean Seattle, WA More articles by this author Robert Welikson Seattle, WA More articles by this author Kathleen Kobashi Seattle, WA More articles by this author James Dennis Seattle, WA More articles by this author Margaret Allen Seattle, WA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...