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Research Papers

Role of Cells in Freezing-Induced Cell-Fluid-Matrix Interactions Within Engineered Tissues

[+] Author and Article Information
Altug Ozcelikkale

School of Mechanical Engineering,
Purdue University,
West Lafayette, IN 47907

Craig Dutton

Department of Aerospace Engineering,
University of Illinois at Urbana-Champaign,
Urbana, IL 61801

Bumsoo Han

School of Mechanical Engineering,
Purdue University,
West Lafayette, IN 47907;
Weldon School of Biomedical Engineering,
Purdue University,
West Lafayette, IN 47907
e-mail: bumsoo@purdue.edu

1Angela Seawright and Altug Ozcelikkale have equally contributed to the present work.

2Corresponding author. Present address: 585 Purdue Mall, West Lafayette, IN 47906.

Contributed by the Bioengineering Division of ASME for publication in the JOURNAL OF BIOMECHANICAL ENGINEERING. Manuscript received June 19, 2012; final manuscript received April 29, 2013; accepted manuscript posted May 16, 2013; published online July 10, 2013. Assoc. Editor: John Bischof.

J Biomech Eng 135(9), 091001 (Jul 10, 2013) (12 pages) Paper No: BIO-12-1240; doi: 10.1115/1.4024571 History: Received June 19, 2012; Revised April 29, 2013; Accepted May 16, 2013

During cryopreservation, ice forms in the extracellular space resulting in freezing-induced deformation of the tissue, which can be detrimental to the extracellular matrix (ECM) microstructure. Meanwhile, cells dehydrate through an osmotically driven process as the intracellular water is transported to the extracellular space, increasing the volume of fluid for freezing. Therefore, this study examines the effects of cellular presence on tissue deformation and investigates the significance of intracellular water transport and cell-ECM interactions in freezing-induced cell-fluid-matrix interactions. Freezing-induced deformation characteristics were examined through cell image deformetry (CID) measurements of collagenous engineered tissues embedded with different concentrations of MCF7 breast cancer cells versus microspheres as their osmotically inactive counterparts. Additionally, the development of a biophysical model relates the freezing-induced expansion of the tissue due to the cellular water transport and the extracellular freezing thermodynamics for further verification. The magnitude of the freezing-induced dilatation was found to be not affected by the cellular water transport for the cell concentrations considered; however, the deformation patterns for different cell concentrations were different suggesting that cell-matrix interactions may have an effect. It was, therefore, determined that intracellular water transport during freezing was insignificant at the current experimental cell concentrations; however, it may be significant at concentrations similar to native tissue. Finally, the cell-matrix interactions provided mechanical support on the ECM to minimize the expansion regions in the tissues during freezing.

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References

Berthiaume, F., Moghe, P. V., Toner, M., and Yarmush, M. L., 1996, “Effect of Extracellular Matrix Topology on Cell Structure, Function, and Physiological Responsiveness: Hepatocytes Cultured in a Sandwich Configuration,” FASEB J., 10, pp. 1471–1484. Available at: http://www.fasebj.org/content/10/13/1471.full.pdf [PubMed]
Borene, M. L., Barocas, V. H., and Hubel, A., 2004, “Mechanical and Cellular Changes During Compaction of a Collagen-Sponge-Based Corneal Stromal Equivalent,” Ann. Biomed. Eng., 32(2), pp. 274–283. [CrossRef] [PubMed]
Meredith, J. E., Fazeli, B., and Schwartz, M. A., 1993, “The Extracellular Matrix as a Cell Survival Factor,” Mol. Biol. Cell, 4, pp. 953–961. [PubMed]
Grinnell, F., 1994, “Fibroblasts, Myofibroblasts, and Wound Contraction,” J. Cell Biol., 124, pp. 401–404. [CrossRef] [PubMed]
Petroll, W. M., and Ma, L., 2003, “Direct, Dynamic Assessment of Cell-Matrix Interactions Inside Fibrillar Collagen Lattices,” Cell Motil. Cytoskeleton, 55, pp. 254–264. [CrossRef] [PubMed]
Gerson, C. J., Goldstein, S., and Heacox, A. E., 2009, “Retained Structural Integrity of Collagen and Elastin Within Cryopreserved Human Heart Valve Tissue as Detected by Two-Photon Laser Scanning Confocal Microscopy,” Cryobiology, 59(2), pp. 171–179. [CrossRef] [PubMed]
Schenke-Layland, K., Xie, J., Heydarkhan-Hagvall, S., Hamm-Alvarez, S. F., Stock, U. A., Brockbank, K. G., and MacLellan, W. R., 2007, “Optimized Preservation of Extracellular Matrix in Cardiac Tissue: Implications for Long-Term Graft Durability,” Ann. Thorac. Surg., 83, pp. 1641–1650. [CrossRef] [PubMed]
Karlsson, J. O., 2010, “Effects of Solution Composition on the Theoretical Prediction of Ice Nucleation Kinetics and Thermodynamics,” Cryobiology, 60(1), pp. 43–51. [CrossRef] [PubMed]
Mazur, P., 1984, “Freezing of Living Cells: Mechanisms and Implications,” Am. J. Physiol., 247(3 Pt 1), pp. C125–142. [PubMed]
Toner, M., Cravalho, E. G., and Karel, M., 1990, “Thermodynamics and Kinetics of Intracellular Ice Formation During Freezing of Biological Cells,” J. Appl. Phys., 67(3), pp. 1582–1593. [CrossRef]
Zhurova, M., Woods, E. J., and Acker, J. P., 2010, “Intracellular Ice Formation in Confluent Monolayers of Human Dental Stem Cells and Membrane Damage,” Cryobiology, 61(1), pp. 133–141. [CrossRef] [PubMed]
Bischof, J. C., Hunt, C. J., Rubinsky, B., Burgess, A., and Pegg, D. E., 1990, “Effects of Cooling Rate and Glycerol Concentration on the Structure of the Frozen Kidney: Assessment by Cryoscanning Electron Microscopy,” Cryobiology, 27, pp. 301–310. [CrossRef] [PubMed]
Hong, J. S., and Rubinsky, B., 1994, “Patterns of Ice Formation in Normal and Malignant Breast Tissue,” Cryobiology, 31(2), pp. 109–120. [CrossRef] [PubMed]
Pazhayannur, P. V., and Bischof, J. C., 1997, “Measurement and Simulation of Water Transport During Freezing in Mammalian Liver Tissue,” ASME J. Biomech. Eng., 119, pp. 269–277. [CrossRef]
Brockbank, K. G., MacLellan, W. R., Xie, J., Hamm-Alvarez, S. F., Chen, Z. Z., and Schenke-Layland, K., 2008, “Quantitative Second Harmonic Generation Imaging of Cartilage Damage,” Cell Tissue Bank., 9(4), pp. 299–307. [CrossRef] [PubMed]
Laouar, L., Fishbein, K., McGann, L. E., Horton, W. E., Spencer, R. G., and Jomha, N. M., 2007, “Cryopreservation of Porcine Articular Cartilage: MRI and Biochemical Results After Different Freezing Protocols,” Cryobiology, 54(1), pp. 36–43. [CrossRef] [PubMed]
Oskam, I. C., Lund, T., and Santos, R. R., 2011, “Irreversible Damage in Ovine Ovarian Tissue After Cryopreservation in Propanediol: Analyses After In Vitro Culture and Xenotransplantation,” Reprod. Domest. Anim., 46(5), pp. 793–799. [CrossRef] [PubMed]
Venkatasubramanian, R. T., Wolkers, W. F., Shenoi, M. M., Barocas, V. H., Lafontaine, D., Soule, C. L., Iaizzo, P. A., and Bischof, J. C., 2010, “Freeze-Thaw Induced Biomechanical Changes in Arteries: Role of Collagen Matrix and Smooth Muscle Cells,” Ann. Biomed. Eng., 38(3), pp. 694–706. [CrossRef] [PubMed]
Changoor, A., Fereydoonzad, L., Yaroshinsky, A., and Buschmann, M. D., 2010, “Effects of Refrigeration and Freezing on the Electromechanical and Biomechanical Properties of Articular Cartilage,” ASME J. Biomech. Eng., 132(6), p. 064502. [CrossRef]
Baicu, S., Taylor, M. J., Chen, Z., and Rabin, Y., 2006, “Vitrification of Carotid Artery Segments: An Integrated Study of Thermophysical Events and Functional Recovery Toward Scale-Up for Clinical Applications,” Cell Preserv. Technol., 4(4), pp. 236–244. [CrossRef] [PubMed]
Dainese, L., Barili, F., Topkara, V. K., Cheema, F. H., Formato, M., Aljaber, E., Fusari, M., Micheli, B., Guarino, A., Biglioli, P., and Polvani, G., 2006, “Effect of Cryopreservation Techniques on Aortic Valve Glycosaminoglycans,” Artif. Organs, 30(4), pp. 259–264. [CrossRef] [PubMed]
Gustavo, M., Andrade, M. G., Sa, C. N., Marchionni, A. M., dos Santos Calmon de Bittencourt, T. C., and Sadigursky, M., 2008, “Effects of Freezing on Bone Histological Morphology,” Cell Tissue Bank., 9(4), pp. 279–287. [CrossRef] [PubMed]
Han, B., Miller, J. D., and Jung, J. K., 2009, “Freezing-Induced Fluid-Matrix Interaction in Poroelastic Material,” ASME J. Biomech. Eng., 131(2), p. 021002. [CrossRef]
Teo, K. Y., DeHoyos, T. O., Dutton, J. C., Grinnell, F., and Han, B., 2011, “Effects of Freezing-Induced Cell-Fluid-Matrix Interactions on the Cells and Extracellular Matrix of Engineered Tissues,” Biomaterials, 32(23), pp. 5380–5390. [CrossRef] [PubMed]
Teo, K. Y., Dutton, J. C., and Han, B., 2010, “Spatiotemporal Measurement of Freezing-Induced Deformation of Engineered Tissues,” ASME J. Biomech. Eng., 132(3), p. 031003. [CrossRef]
Noel, A., Munaut, C., Boulvain, A., Calberg-Bacq, C. M., Lambert, C. A., Nusgens, B., Lapiere, C. M., and Foidart, J. M., 1992, “Modulation of Collagen and Fibronectin Synthesis in Fibroblasts by Normal and Malignant Cells,” J. Cell. Biochem., 48, pp. 150–161. [CrossRef] [PubMed]
Devireddy, R. V., Swanlund, D. J., Roberts, K. P., and Bischof, J. C., 1999, “Subzero Water Permeability Parameters of Mouse Spermatozoa in the Presence of Extracellular Ice and Cryoprotective Agents,” Biol. Reprod., 61(3), pp. 764–775. [CrossRef] [PubMed]
Balasubramanian, S. K., Bischof, J. C., and Hubel, A., 2006, “Water Transport and IIF Parameters for a Connective Tissue Equivalent,” Cryobiology, 52(1), pp. 62–73. [CrossRef] [PubMed]
Akhoondi, M., Oldenhof, H., Stoll, C., Sieme, H., and Wolkers, W. F., 2011, “Membrane Hydraulic Permeability Changes During Cooling of Mammalian Cells,” Biochim. Biophys. Acta, 1808(3), pp. 642–648. [CrossRef] [PubMed]
Devireddy, R. V., Raha, D., and Bischof, J. C., 1998, “Measurement of Water Transport During Freezing in Cell Suspensions Using a Differential Scanning Calorimeter,” Cryobiology, 36(2), pp. 124–155. [CrossRef] [PubMed]
Toner, M., Tompkins, R. G., Cravalho, E. G., and Yarmush, M. L., 1992, “Transport Phenomena During Freezing of Isolated Hepatocytes,” AIChE J., 38(10), pp. 1512–1522. [CrossRef]
Mazur, P., 1963, “Kinetics of Water Loss From Cells at Subzero Temperatures and Likelihood of Intracellular Freezing,” J. Gen. Physiol., 47, pp. 347–369. [CrossRef] [PubMed]
Levin, R. L., Cravalho, E. G., and Huggins, C. E., 1976, “A Membrane Model Describing the Effect of Temperature on the Water Conductivity of Erythrocyte Membranes at Subzero Temperatures,” Cryobiology, 13(4), pp. 415–429. [CrossRef] [PubMed]
Levenberg, K., 1944, “A Method for the Solution of Certain Non-Linear Problems in Least Squares,” Q. Appl. Math., 2, pp. 164–168.
Pazhayannur, P. V., and Bischof, J. C., 1997, “Measurement and Simulation of Water Transport During Freezing in Mammalian Liver Tissue,” ASME J. Biomech. Eng., 119(3), pp. 269–277. [CrossRef]
Devireddy, R. V., Smith, D. J., and Bischof, J. C., 2002, “Effect of Microscale Mass Transport and Phase Change on Numerical Prediction of Freezing in Biological Tissues,” ASME J. Heat Transfer, 124(2), pp. 365–374. [CrossRef]
Devireddy, R. V., Thirumala, S., and Gimble, J. M., 2005, “Cellular Response of Adipose Derived Passage-4 Adult Stem Cells to Freezing Stress,” ASME J. Biomech. Eng., 127(7), pp. 1081–1086. [CrossRef]
Ivascu, A., and Kubbies, M., 2007, “Diversity of Cell-Mediated Adhesions in Breast Cancer Spheroids,” Int. J. Oncol., 31, pp. 1403–1413. [PubMed]
Viskanta, R., Bianchi, M. V. A., Critser, J. K., and Gao, D., 1997, “Solidification Processes of Solutions,” Cryobiology, 34(4), pp. 348–362. [CrossRef] [PubMed]
Wright, J., Han, B., and Chuong, C. J., 2012, “Biphasic Investigation of Tissue Mechanical Response During Freezing Front Propagation,” ASME J. Biomech. Eng., 134(6), p. 061005. [CrossRef]
He, X., and Bischof, J. C., 2005, “Analysis of Thermal Stress in Cryosurgery of Kidneys,” ASME J. Biomech. Eng., 127(4), pp. 656–661. [CrossRef]
Shi, X., Datta, A. K., and Mukherjee, Y., 1998, “Thermal Stresses From Large Volumetric Expansion During Freezing of Biomaterials,” ASME J. Biomech. Eng., 120(6), pp. 720–726. [CrossRef]
Pitt, R. E., 1990, “Cryobiological Implications of Different Methods of Calculating the Chemical-Potential of Water in Partially Frozen Suspending Media,” CryoLetters, 11(3), pp. 227–240.
Swaminathan, C. R., and Voller, V. R., 1992, “A General Enthalpy Method for Modeling Solidification Processes,” Metall. Trans. B, 23(5), pp. 651–664. [CrossRef]
Ahmadikia, H., and Moradi, A., 2012, “Non-Fourier Phase Change Heat Transfer in Biological Tissues During Solidification,” Heat Mass Transfer, 48(9), pp. 1559–1568. [CrossRef]
Choi, J., and Bischof, J. C., 2011, “Cooling Rate Dependent Biophysical and Viability Response Shift With Attachment State in Human Dermal Fibroblast Cells,” Cryobiology, 63(3), pp. 285–291. [CrossRef] [PubMed]
Balasubramanian, S. K., Wolkers, W. F., and Bischof, J. C., 2009, “Membrane Hydration Correlates to Cellular Biophysics During Freezing in Mammalian Cells,” Biochim. Biophys. Acta, 1788(5), pp. 945–953. [CrossRef] [PubMed]
Wolkers, W. F., Balasubramanian, S. K., Ongstad, E. L., Zec, H. C., and Bischof, J. C., 2007, “Effects of Freezing on Membranes and Proteins in LNCaP Prostate Tumor Cells,” Biochim. Biophys. Acta, 1768(3), pp. 728–736. [CrossRef] [PubMed]
Kleinhans, F. W., and Mazur, P., 2009, “Determination of the Water Permeability (Lp) of Mouse Oocytes at −25 Degrees C and its Activation Energy at Subzero Temperatures,” Cryobiology, 58(2), pp. 215–224. [CrossRef] [PubMed]
Yang, G., Veres, M., Szalai, G., Zhang, A. L., Xu, L. X., and He, X. M., 2011, “Biotransport Phenomena in Freezing Mammalian Oocytes,” Ann. Biomed. Eng., 39(1), pp. 580–591. [CrossRef] [PubMed]
Smith, D. J., Schulte, M., and Bischof, J. C., 1998, “The Effect of Dimethylsulfoxide on the Water Transport Response of Rat Hepatocytes During Freezing,” ASME J. Biomech. Eng., 120(5), pp. 549–558. [CrossRef]
Quinn, T. M., Hunziker, E. B., and Hauselmann, H. J., 2005, “Variation of Cell and Matrix Morphologies in Articular Cartilage Among Locations in the Adult Human Knee,” Osteoarthritis Cartilage, 13(8), pp. 672–678. [CrossRef] [PubMed]
Sander, E. A., and Barocas, V. H., 2008, “Biomimetic Collagen Tissues: Collagenous Tissue Engineering and Other Applications,” Collagen, P.Fratzl, ed., Springer, New York, pp. 475–504.
Vunjak-Novakovic, G., Tandon, N., Godier, A., Maidhof, R., Marsano, A., Martens, T. P., and Radisic, M., 2010, “Challenges in Cardiac Tissue Engineering,” Tissue Eng. Part B, Rev., 16(2), pp. 169–187. [CrossRef]

Figures

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Fig. 1

MCF7 cells (left column, (a) and (c)) and Fluoresbrite® YG Microspheres (right column (b) and (d)) embedded in engineered tissues. Images (a) (MCF7 cells) and (b) (microspheres) are imaged with the bright field of a confocal microscope (Olympus IX71). Images (c) and (d) are fluorescent images, the MCF7 embedded QDs, (c), imaged with the TRITC filter and the microspheres, (d), imaged with the FITC filter.

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Fig. 2

Schematic illustration of the representative elementary volume of an engineered tissue. States (I) and (II) correspond to before and during extracellular freezing. The ET is composed of cellular (c) and extracellular (ec) compartments. Each compartment has both solid (s) and fluid (fl) components. As ice forms in the extracellular space, cells dehydrate and make more water available for extracellular freezing. In the meantime, the extracellular space expands increasing the volume of the REV.

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Fig. 3

Dilatation (n ≥ 3) of normal MCF7 concentration (a) and microsphere ETs (b) at X(t) = 2000 μm

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Fig. 4

The y-averaged deformation (top row) and dilatation rates (bottom row) at X(t) = 2000 μm and 4000 μm for (a) and (c) the normal MCF7 cell concentration and (b) and (d) the microspheres

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Fig. 5

Dilatation (n ≥ 3) of doubled MCF7 concentration (a) and quadrupled MCF7 concentration ETs (b) at X(t) = 2000 μm

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Fig. 6

The deformation (top row) and dilatation rates (bottom row) at X(t) = 2000 μm and 4000 μm averaged along the y-axis for (a) and (c) the doubled MCF7 cell concentration and (b) and (d) the quadrupled MCF7 cell concentration

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Fig. 7

Cellular water transport during freezing. (a) Representative cryomicroscopy images of MCF7 cells dehydrating during freezing with the cooling rate of 10  °C/min. The locations of cells are indicated by dark arrows. Scale bar is 20 μm (b) change of cell volume with decreasing temperature for the three cooling rates. The error bars stand for the standard error of the mean. Solid lines are model predictions with the optimal membrane permeability parameters.

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Fig. 8

Extracellular freezing thermodynamics of engineered tissues. Rate of latent heat release (solid line) and the frozen fraction calculated by cumulative integration of the rate of latent heat release with respect to time (dashed line).

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Fig. 9

(a) Temperature measurements obtained at different axial distances from the cold terminal. The underlying lines are polynomial curve fits that were used in the analysis for the estimation of freezing-induced dilatation (Pearson coefficient of correlation R2 > 0.99 for each case). The temporal trends of (b) cellular water transport, (c) extracellular ice formation, and (d) estimated freezing-induced dilatation at two axial locations, x = 2000 μm and x = 4000 μm.

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Fig. 10

(a) The variation of estimated freezing-induced dilatation with decreasing temperature for different concentrations of cells and microspheres. Solid lines correspond to ETs with cells while dashed lines correspond to ETs with microspheres. (b) The variation of relative difference of freezing-induced dilatation between the cells versus microsphere ETs with decreasing temperature. Unlike the actual experimental settings, this analysis assumes the sphere diameter to be the same as the diameter of the MCF7 cells in order to isolate the effects of cellular water transport from differential size effects.

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