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Prevascularized Acellular Construct for Tissue Engineering

Prevascularized Acellular Construct for Tissue Engineering. Ryan Nagao Apr 9, 2010. Objective. Create a construct that can be connected to the host vascular system for rapid anastomosis and tissue survival for large defects. Current Solutions to Vascularization.

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Prevascularized Acellular Construct for Tissue Engineering

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  1. PrevascularizedAcellular Construct for Tissue Engineering Ryan Nagao Apr 9, 2010

  2. Objective • Create a construct that can be connected to the host vascular system for rapid anastomosis and tissue survival for large defects

  3. Current Solutions to Vascularization Jain RK., et al. Nat Biotechnol, 2005

  4. PrevascularizingAcellular Tissue in vitro Decellularization EC cell seeding Excision Perfusion culture

  5. Specific Aims

  6. Specific Aims

  7. Specific Aim 1 • Specific Aim 1 – Preservation of intricate microvasculature using decellularization • Evaluate the scaffold composition for cellularity and ECM components against unmodified tissue from the same source • Implantation subcutaneously to evaluate immunogenicity in vivo

  8. Aim 1: Experimental Strategy neurophilosophy.files.wordpress.com/ Tom WJ., et al. IEEE T Med Imaging, 2008

  9. Previously…

  10. Preliminary Research—Presence of CD-31 Following Decell Low concentration DAPI CD-31 High concentration Acell Lung Fresh Lung

  11. Preliminary Research—Preservation of ECM Proteins Following High Concentration Decell DAPI FN LN Acell Lung Fresh Lung

  12. Speckle Imaging • Gives relative flow velocities • Used by Dunn’s lab to visualize occlusions and opened pathways in the brain • Modify resolution and depth of field using different objectives • I can determine the patency following decellularization using this method Tom WJ., et al. IEEE T Med Imaging, 2008

  13. Fractal Analysis • Way to measure geometries that are not of integer order • Describes self similarity • Describes complexity http://www.proetcontra.com/wp-content/uploads/fractal_geometry.jpg

  14. What Am I Talking About? • Consider • Increasing L by 2 results in 4 similar objects • Follows a power law • Dim=2, thus a triangle is considered a 2 dimensional figure yay! Length = L Length = 2L N= scaledim 4= 2dim

  15. Calculating a Fractal Dimension • Now consider • 2dim=3 • Dim=ln(3)/ln(2) = 1.58 http://scienceblogs.com/goodmath/2007/08/fractal_dimension_1.php

  16. Fractal Dimensions are Present in Nature • Trees, coastlines, and vascular beds have all been characterized using fractal analysis • Vessels, Df = ~1.7 • Capillaries, Df = ~2 • Tumors, Df = ~1.9 • Implement a box-counting algorithm on a skeletonized figure • This yields regressions lines whose slope will give the fractal dimension of your figure • Can also get information about branching from the skeletonized image through an analysis algorithm

  17. Fractal Image Processing

  18. Perfusion System Calculations • Shear calculation • My culture system *Inoguchi H., et al. Biomaterials, 2006

  19. Flow System Set-up • Flow rates will be measured real-time using Doppler ultrasound • Shear rates can be determined based off of flow rates • Side project could determine the benefits that acellular vascular ECM imparts on endothelial cell alignment 1 mm Pressure control valve Reservoir T T Incubator

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