0
Research Papers

Investigation of the In Vitro Culture Process for Skeletal-Tissue-Engineered Constructs Using Computational Fluid Dynamics and Experimental Methods

[+] Author and Article Information
Md. Shakhawath Hossain

e-mail: mdh511@mail.usask.ca

X. B. Chen

e-mail: xbc719@mail.usask.ca

D. J. Bergstrom

e-mail: don.bergstrom@usask.ca
Department of Mechanical Engineering,
University of Saskatchewan,
57 Campus Drive Saskatoon, SK,
S7N 5A9, Canada

1Corresponding author.

Contributed by the Bioengineering Division of ASME for publication in the JOURNAL OF BIOMECHANICAL ENGINEERING. Manuscript received May 23, 2012; final manuscript received October 4, 2012; accepted manuscript posted October 25, 2012; published online November 27, 2012. Assoc. Editor: Pasquale Vena.

J Biomech Eng 134(12), 121003 (Nov 27, 2012) (11 pages) doi:10.1115/1.4007952 History: Received May 23, 2012; Revised October 04, 2012; Accepted October 25, 2012

The in vitro culture process via bioreactors is critical to create tissue-engineered constructs (TECs) to repair or replace the damaged tissues/organs in various engineered applications. In the past, the TEC culture process was typically treated as a black box and performed on the basis of trial and error. Recently, computational fluid dynamics (CFD) has demonstrated its potential to analyze the fluid flow inside and around the TECs, therefore, being able to provide insight into the culture process, such as information on the velocity field and shear stress distribution that can significantly affect such cellular activities as cell viability and proliferation during the culture process. This paper briefly reviews the CFD and experimental methods used to investigate the in vitro culture process of skeletal-type TECs in bioreactors, where mechanical deformation of the TEC can be ignored. Specifically, this paper presents CFD modeling approaches for the analysis of the velocity and shear stress fields, mass transfer, and cell growth during the culture process and also describes various particle image velocimetry (PIV) based experimental methods to measure the velocity and shear stress in the in vitro culture process. Some key issues and challenges are also identified and discussed along with recommendations for future research.

FIGURES IN THIS ARTICLE
<>
Copyright © 2012 by ASME
Your Session has timed out. Please sign back in to continue.

References

Blitterswijk, C. V., 2008, Tissue Engineering, Elsevier Inc., London.
Meyer, U., Meyer, T., Handschel, J., and Wiesmann, H. P., 2009, Fundamentals of Tissue Engineering and Regenerative Medicine, Springer-Verlag Berlin Heidelberg.
Milan, J., Planell, J. S., and Lacroix, D., 2010, “Simulation of Bone Tissue Formation Within a Porous Scaffold Under Dynamic Compression,” Biomech. Model. Mechan., 9, pp. 583–596. [CrossRef]
Porter, B., Zauel, R., Stockman, H., Guldberg, R., and Fyhrie, D., 2005, “3-D Computational Modeling of Media Flow Through Scaffolds in a Perfusion Bioreactor,” J. Biomech., 38, pp. 543–549. [CrossRef] [PubMed]
Xie, Y. Z., Hardouin, P., Zhu, Z. N., Tang, T. T., Dai, K. R., and Lu, J. X., 2006, “Three-Dimensional Flow Perfusion Culture System for Stem Cell Proliferation Inside the Critical-Size Betatricalcium Phosphate Scaffold,” Tissue Eng., 12, pp. 3535–3543. [CrossRef] [PubMed]
Li, D., Tang, T., Lu, J., and Dai, K., 2009, “Effects of Flow Shear Stress and Mass Transport on the Construction of a Large-Scale Tissue-Engineered Bone in a Perfusion Bioreactor,” Tissue Eng. A, 15, pp. 2773–2783. [CrossRef]
Yeatts, A. B., and Fisher, J. P., 2011, “Bone Tissue Engineering Bioreactors: Dynamic Culture and Influence of Shear Stress,” Bone, 48, pp. 171–181. [CrossRef] [PubMed]
Singh, H., and Hutmacher, D. W., 2009, “Bioreactor Studies and Computational Fluid Dynamics,” Adv. Biochem. Eng. Biotech., 112, pp. 231–249.
Fraser, K. H., Taskin, M. E., Griffith, B. P., and Wu, Z. J., 2011, “The Use of Computational Fluid Dynamics in the Development of Ventricular assists Devices,” Med. Eng. Physics, 33, pp. 263–280. [CrossRef]
Betchen, L. J., and Straatman, A.G., 2010, “An Investigation of the Effects of a Linear Porosity Distribution on Non-Equilibrium Heat Transfer in High-Conductivity Graphite Foam,” Num. Heat Transf. A, 58, pp. 605–624. [CrossRef]
Djilali, N., 2007, “Computational Modeling of Polymer Electrode Membrane (PEM) Fuel Cells: Challenges and Opportunity,” Energy, 32, pp. 269–280. [CrossRef]
Sozer, E., and ShyyW., 2007, “Modeling of Fluid Dynamics and Heat Transfer Through Porous Media for Rocket Propulsion,” 43rd AIAA/ASME/SAE/ASEE Joint Propulsion Conference & Exhibit, Cincinnati, OH.
Voronov, R., VanGordon, S., Sikavitsas, V., and Papavassiliou, D., 2010, “Computational Modeling of Flow-Induced Shear Stresses Within 3D Salt-Leached Porous Scaffolds Imaged via Micro-CT,” J. Biomech., 43, pp. 1279–1286. [CrossRef] [PubMed]
Patankar, S. V., 1980, Numerical Heat Transfer and Fluid Flow, McGraw-Hill, New York.
Ferziger, J. H., and Peric, M., 2002, Computational Methods for Fluid Dynamics, Springer, New York.
Darcy, H., 1856, Les fontaines publiques de la ville de diljon, Dalmont, Paris.
Brinkman, H. C., 1949, “On the Permeability of Media Consisting of Closely Packed Porous Particles,” Appl. Sci. Res., A1, pp. 81–86. [CrossRef]
Whitaker, S., 1986, “Flow in Porous Media I: A Theoretical Derivation of Darcy's Law,” Transport Porous Med., 1(1), pp. 3–25. [CrossRef]
Chung, C. A., Chen, C. W., Chen, C. P., and Tseng, C. S., 2007, “Enhancement of Cell Growth in Tissue-Engineering Constructs Under Perfusion Modeling and Simulation,” Biotech. Bioeng., 97(6), pp. 1603–1616. [CrossRef]
Golfier, F., Wood, B. D., Orgogozo, L., Quintard, M., and Bues, M., 2009, “Biofilms in Porous Media: Development of Macroscopic Transport Equations via Volume Averaging With Closure for Local Mass Equilibrium Conditions,” Adv. Water Resour., 32, pp. 463–485. [CrossRef]
Boccaccio, A., Ballini, A., Pappalettere, C., Tullo, D., Cantore, S., and Desiate, A., 2011, “Finite Element Method (FEM), Mechanobiology and Biomimetic Scaffolds in Bone Tissue Engineering,” Int. J. Biolog. Sci., 7(1), pp. 112–132. [CrossRef]
Lacroix, D., Prendergast, P. J., Li, G., and Marsh, D., 2002, “Biomechanical Model to Simulate Tissue Differentiation and Bone Regeneration: Application to Fracture Healing,” Med. Biol. Eng. Comp., 40, pp. 14–21. [CrossRef]
Shakeel, M., 2011, “Continuum Modeling of Cell growth and Nutrient Transport in a Perfusion Bioreactor,” Ph.D. thesis, School of Mathematics and IT Department, University of Nottingham, Nottingham, U.K.
Singh, H., Teoh, S. H., Low, H. T., and Hutmacher, D. W, 2005, “Flow Modeling Within a Scaffold Under the Influence of Uni-Axial and Bi-Axial Bioreactor Reaction,” J. Biotech., 119, pp. 181–196. [CrossRef]
Sacco, R., Causin, P., Zunino, P., and Raimondi, M. T., 2011, “A Multiphysics/Multiscale 2D Numerical Simulation of Scaffold-Based Cartilage Regeneration Under Interstitial Perfusion in a Bioreactor,” Biomech. Model. Mechan., 10(4), pp. 577–589. [CrossRef]
Raimondi, M. T., Causin, P.Mara, A., Nava, M., Laganà, M., and Sacco, R., 2011, “Breakthroughs in Computational Modeling of Cartilage Regeneration in Perfused Bioreactors,” IEEE T. Biomed. Eng., 58(12), pp. 3496–3499. [CrossRef]
Pauwels, F., 1960, “A New Theory on the Influence of Mechanical Stimuli on the Differentiation of Supporting Tissue. The Tenth Contribution to the Functional Anatomy and Causal Morphology of the Supporting Structure,” Z. Anat. Entwickl. Gesch., 121, pp. 478–515. [CrossRef]
Claes, L., and Heigele, C., 1999, “Magnitudes of Local Stress and Strain Along Bony Surfaces Predicts the Course and Type of Fracture Healing,” J. Biomech., 32, pp. 255–266. [CrossRef] [PubMed]
Isakssona, H., Comas, O., Donkelaar, C., Mediavilla, J., Wilsonb, W., Huiskesb, R., and Itoa, K., 2006, “Bone Regeneration During Distraction Osteogenesis: Mechano-Regulation by Shear Strain and Fluid Velocity,” J. Biomech., 35, pp. 2002–2011.
Prendergast, P. J., and Lacroix, D., 2002, “A Mechano-Regulation Model for Tissue Differentiation During Fracture Healing: Analysis of Gap Size and Loading,” J. Biomech., 35, pp. 1163–1171. [CrossRef] [PubMed]
Khayyeri, H., Checa, S., Tagil, M., and Prendergast, P. J., 2009, “Corroboration of Mechanobiological Simulations of Tissue Differentiation in an in vivo Bone Chamber Using a Lattice-Modeling Approach,” J. Orthoped. Res., 27(12), pp. 1659–1666. [CrossRef]
Tagil, M., and Aspenberg, P., 1999, “Cartilage Induction by Controlled Mechanical Stimulation in vivo,” J. Orthoped. Res., 17, pp. 200–204. [CrossRef]
Bottaro, D. P., Liebmann-Vinson, A., and Heidaran, M. A., 2002 “Molecular Signaling in Bioengineered Tissue Microenvironment,” Ann. N.Y. Acad. Sci., 961, pp. 143–153. [CrossRef] [PubMed]
Korossis, S. A., Bolland, F., Kearney, J. N., Fisher, J., and Ingham, E., 2005, “Bioreactors in Tissue Engineering,” Top. Tissue Eng., 5, pp. 1–24.
McCoy, R. J., Jungreuthmayer, C., and O'Brien, F. J., 2012, “Influence of Flow Rate and Scaffold Pore Size on Cell Behavior During Mechanical Simulation in Flow Perfusion Bioreactor,” Biotech. Bioeng., 109(6), pp. 1583–1594. [CrossRef]
Hidalgo-Bastida, L. A., Thirunavukkarasu, S., Griffiths, S., Cartmell, S. H., and Naire, S., 2012, “Modeling and Design of Optimal Flow Perfusion Bioreactors for Tissue Engineering Applications,” Biotech. Bioeng., 109(4), pp. 1095–1099. [CrossRef]
Bilgen, B., and Barabino, G. A., 2007, “Location of Scaffolds in Bioreactors Modulates the Hydrodynamic Environment Experienced by Engineered Tissues,” Biotech. Bioeng., 98(1), pp. 282–294. [CrossRef]
Gutierrez, R. A., and Crumpler, E. T., 2007, “Potential Effect of Geometry on Wall Shears Stress Distribution Across Scaffold Surfaces,” Ann. Biomed. Eng., 36(1), pp. 77–85. [CrossRef] [PubMed]
Lanza, R. P., Langer, R., and Vacanti, J., 2000, Principles of Tissue Engineering, Academic Press, San Diego, CA.
Vunjak-Novakovic, G., Freed, L. E., Biron, R. J., and Langer, R., 2004, “Effects of Mixing on Tissue Engineered Cartilage,” AIChE J., 42(3), pp. 850–860. [CrossRef]
Singh, H., Ang, E. S., Lim, T. T., and Hutmacher, D.W., 2006, “Flow Modeling in a Novel Non-Perfusion Conical Bioreactor,” Biotech. Bioeng., 97(5), pp. 1291–1299. [CrossRef]
Bancroft, G. N., Sikavitsas, V. I., and Mikos, A. G., 2003, “Design of a Flow Perfusion Bioreactor System for Bone Tissue Engineering Application,” Tissue Eng., 9(3), pp. 549–554. [CrossRef] [PubMed]
Cioffi, M., Boschetti, F., Raimondi, M. T., and Dubini, G., 2006, “Modelling Evaluation of the Fluid-Dynamic Microenvironment in Tissue-Engineered Constructs: A Micro-CT Based Model,” Biotech. Bioeng., 93(3), pp. 500–510. [CrossRef]
Raimondi, M. T., Boschetti, F., Falcone, L., Migliavacca, F., Remuzzi, A., and Dubini, G., 2004, “The Effect of Media Perfusion on Three-Dimensional Cultures of Human Chondrocytes: Integration of Experimental and Computational Approaches,” Biorheology41, pp. 401–410. [PubMed]
Bruneau, C-H., and Mortazavi, I., 2008, “Numerical Modeling and Passive Flow Control Using Porous Media,” Comput. Fluids, 37, pp. 488–498. [CrossRef]
Cheng, G., Youssef, B. B., Markenscoff, P., and Zygourakis, K., 2006, “Cell Population Dynamics Modulate the Rates of Tissue Growth Processes,” Biophys. J., 90(3), pp. 713–724. [CrossRef] [PubMed]
Galbusera, F., Cioffi, M., Raimondi, M. T., and Pietrabissa, R., 2007, “Computational Modeling of Combined Cell Population Dynamics and Oxygen Transport in Engineered Tissue Subject to Interstitial Perfusion” Comput. Method. Biomec., 10(4), pp. 279–287. [CrossRef]
Lemon, G., and KingJ. R., 2007, “Multiphase Modeling of Cell Behavior on Artificial Scaffolds: Effects of Nutrient Depletion and Spatially Non-Uniform Porosity,” Math. Med. Biol., 24(1), pp. 57–83. [CrossRef] [PubMed]
Laganà, M., and Raimondi, M. T., 2012, “A Miniaturized, Optically Accessible Bioreactor for Systematic 3D Tissue Engineering Research,” Biomed. Microdevices, 14(1), pp. 225–234. [CrossRef] [PubMed]
Adrian, R. J., “Twenty Years of Particle Image Velocimetry,” Exp. Fluids, 39, pp. 159–169. [CrossRef]
Adrian, R. J., 1997, “Dynamic Ranges of Velocity and Spatial Resolution of Particle Image Velocimetry,” Meas. Sci. Technol., 8, pp. 1393–1398. [CrossRef]
Raffel, M., Willert, C., and Kompenhans, J., 1998, Particle Image Velocimetry—A Practical Guide, Springer, New York.
Northrup, M. A., Kulp, T. J., and Angel, S. M., 1991, “Fluorescent Particle Image Velocimetry: Application to Flow Measurement in Refractive Index-Matched Porous Media,” Appl. Optics, 30(1), pp. 3034–3040. [CrossRef]
Peurrung, L. M., Rashidi, M., and Kulp, T. J., 1995, “Measurement of Porous Medium Velocity Fields and Their Volumetric Averaging Characteristics Using Particle Tracking Velocimetry,” Chem. Eng. Sci., 50(14), pp. 2243–2253. [CrossRef]
Northrup, M. A., Kulp, T. J., and Angel, S. M., 1991, “Application of Fluorescent Particle Imaging to Measuring Flow in Complex Media,” Anal. Chim. Acta., 255, pp. 275–282. [CrossRef]
Bown, M. R., MacInnes, J. M., Allen, R. W. K., and Zimmerman, W. B. J., 2006, “Three-Dimensional Velocity Measurements Using Stereoscopic Micro-PIV and PTV,” Meas. Sci. Technol., 17, pp. 2175–2185. [CrossRef]
Dusting, J., Sheridan, J., and Hourigan, K., 2006, “A Fluid Dynamics Approach to Bioreactor Design for Cell and Tissue Culture,” Biotech. Bioeng., 94(6), pp. 1196–1208. [CrossRef]
Sucosky, P., Osorio, D. F., Brown, J. B., and Neitzel, G. P., 2004, “Fluid Mechanics of a Spinner-Flask Bioreactor,” Biotech. Bioeng., 85(1), pp. 34–46. [CrossRef]
Santiago, J. G., Wereley, S. T., Meinhart, C. D., Beebe, D. J., and Adrian, R. J., 1998, “A Particle Image Velocimetry System for Microfluidics,” Exp. Fluids, 25(4), pp. 316–319. [CrossRef]
Mielnik, M. M., and Saetran, L. R., 2004, “Micro Particle Image Velocimetry—An Overview,” Turbulence, 10, pp. 83–90.
Meinhart, C. D., Wereley, S. T., and Gray, M. H. B., 2000, “Volume Illumination for Two Dimensional Particle Image Velocimetry,” Meas. Sci. Technol., 11, pp. 809–814. [CrossRef]
Lima, R., Wada, S., Tanaka, S., Takeda, M., Ishikawa, T., Tsubota, K., Imai, Y., and Yamaguchi, T., 2008, “in vitro Blood Flow in a Rectangular PDMS Microchannel: Experimental Observations Using a Confocal; Micro-PIV System,” Biomed. Microdevices, 10(2), pp. 153–167. [CrossRef] [PubMed]
Provin, C., Takano, K., Sakai, Y., Fujii, T., and Shirakashi, R., 2008, “A Method for the Design of 3D Scaffolds for High-Density Cell Attachment and Determination of Optimum Perfusion Culture Conditions,” J. Biomech., 41, pp. 1436–1449. [CrossRef] [PubMed]
De Boodt, S., Truscello, S., Zcan, S., Leroy, T., Van Oosterwyck, H., Berckmans, D., and Schrooten, J., 2010, “Bi-Modular Flow Characterization in Tissue Engineering Scaffolds Using Computational Fluid Dynamics and Particle Imaging Velocimetry,” Tissue Eng. C, 16(6), pp. 1553–1564. [CrossRef]
Kim, G. B., Je, J. H., and Lee, S. J., 2007, “Synchrotron X-ray PIV Technique for Measurement of Blood Flow Velocity,” Synchrotron Radiation Instrumentation: Ninth International Conference [American Institute of Physics, 879, pp. 1891–1894 (2007)].
Fouras, A., Dusting, J., Lewis, R., and Hourigan, K., 2007, “Three-Dimensional Synchrotron X-ray Particle Image Velocimetry,” J. Appl. Phys., 102(6), pp. 064916(1–6). [CrossRef]
Jia, Y., Bagnaninchib, P. O., Yang, Y., Haj, A. E., Hinds, M. T., Kirkpatrick, S. J., and Wang, R. K, 2009, “Doppler Optical Coherence Tomography Imaging of Local Fluid Flow and Shear Stress Within Micro Porous Scaffolds,” J. Biomed. Optics, 14(3), p. 034014. [CrossRef]

Figures

Grahic Jump Location
Fig. 1

A representative elementary volume in the rigid porous medium

Grahic Jump Location
Fig. 2

Schematic diagram of cell growth model

Grahic Jump Location
Fig. 3

Map of local shear stresses (Pa) in media transversely perfused through a 3D trabecular bone TEC from side and top view [4]

Grahic Jump Location
Fig. 4

Contours of macroscopic viscous shear stresses (mPa) after 30 days of culture, at a Peclet number of 100 and 20 μm/s perfusion rates [19]

Grahic Jump Location
Fig. 5

Annotated schematic of the SPIV configuration, viewed from above [57]

Grahic Jump Location
Fig. 6

Schematic of a closed perfusion system used for micro-PIV experiments, consisting of a reservoir, a peristaltic pump, and a bioreactor connected by tubing [64]

Tables

Errata

Discussions

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging and repositioning the boxes below.

Related Journal Articles
Related eBook Content
Topic Collections

Sorry! You do not have access to this content. For assistance or to subscribe, please contact us:

  • TELEPHONE: 1-800-843-2763 (Toll-free in the USA)
  • EMAIL: asmedigitalcollection@asme.org
Sign In