Skip to main content
SpringerLink
Log in

Freeze–Thaw Induced Biomechanical Changes in Arteries: Role of Collagen Matrix and Smooth Muscle Cells

  • Published:
Annals of Biomedical Engineering Aims and scope Submit manuscript

Abstract

Applications involving freeze–thaw, such as cryoplasty or cryopreservation can significantly alter artery biomechanics including an increase in physiological elastic modulus. Since artery biomechanics plays a significant role in hemodynamics, it is important to understand the mechanisms underlying these changes to be able to help control the biomechanical outcome post-treatments. Understanding of these mechanisms requires investigation of the freeze–thaw effect on arterial components (collagen, smooth muscle cells or SMCs), as well as the components’ contribution to the overall artery biomechanics. To do this, isolated fresh swine arteries were subjected to thermal (freeze–thaw to −20 °C for 2 min or hyperthermia to 43 °C for 2 h) and osmotic (0.1–0.2 M mannitol) treatments; these treatments preferentially altered either the collagen matrix (hydration/stability) or smooth muscle cells (SMCs), respectively. Tissue dehydration, thermal stability and SMC functional changes were assessed from bulk weight measurements, analyses of the thermal denaturation profiles using Fourier transform infrared (FTIR) spectroscopy and in vitro arterial contraction/relaxation responses to norepinephrine (NE) and acetylcholine (AC), respectively. Additionally, Second Harmonic Generation (SHG) microscopy was performed on fresh and frozen–thawed arteries to directly visualize the changes in collagen matrix following freeze–thaw. Finally, the overall artery biomechanics was studied by assessing responses to uniaxial tensile testing. Freeze–thaw of arteries caused: (a) tissue dehydration (15% weight reduction), (b) increase in thermal stability (~6.4 °C increase in denaturation onset temperature), (c) altered matrix arrangement observed using SHG and d) complete SMC destruction. While hyperthermia treatment also caused complete SMC destruction, no tissue dehydration was observed. On the other hand, while 0.2 M mannitol treatment significantly increased the thermal stability (~4.8 °C increase in denaturation onset), 0.1 M mannitol treatment did not result in any significant change. Both 0.1 and 0.2 M treatments caused no change in SMC function. Finally, freeze–thaw (506 ± 159 kPa), hyperthermia (268 ± 132 kPa) and 0.2 M mannitol (304 ± 125 kPa) treatments all caused significant increase in the physiological elastic modulus (E artery) compared to control (185 ± 92 kPa) with the freeze–thaw resulting in the highest modulus. These studies suggest that changes in collagen matrix arrangement due to dehydration as well as SMC destruction occurring during freeze–thaw are important mechanisms of freeze–thaw induced biomechanical changes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

Notes

  1. Refers to freeze–thaw data added from our previous publication for comparison.

References

  1. Aksan, A., and J. J. McGrath. Thermomechanical analysis of soft-tissue thermotherapy. J. Biomech. Eng. 125:700–708, 2003.

    Article  PubMed  Google Scholar 

  2. Balasubramanian, S. K., et al. Thermal injury prediction during cryoplasty through in vitro characterization of smooth muscle cell biophysics and viability. Ann. Biomed. Eng. 36(1):86–101, 2008.

    Article  PubMed  Google Scholar 

  3. Bateson, E. A. J., and D. E. Pegg. Cryopreservation of arteries. Cryo-Letters 15:15–20, 1994.

    Google Scholar 

  4. Cai, S., and B. R. Singh. Identification of β-turn and random coil amide III infrared bands for secondary structure estimation of proteins. Biophys. Chem. 80(1):7–20, 1999.

    Article  PubMed  CAS  Google Scholar 

  5. Cai, S., and B. R. Singh. A distinct utility of the amide III infrared band for secondary structure estimation of aqueous protein solutions using partial least-squares methods. Biochemistry 43(9):2541–2549, 2004.

    Article  PubMed  CAS  Google Scholar 

  6. Castier, Y., et al. Cryopreserved arterial allograft reconstruction for peripheral graft infection. J. Vasc. Surg. 41(1):30–37, 2005.

    Article  PubMed  Google Scholar 

  7. Fava, M., et al. Cryoplasty for femoropopliteal arterial disease: late angiographic results of initial human experience. J. Vasc. Interv. Radiol. 15(11):1239–1243, 2004.

    PubMed  Google Scholar 

  8. Fischer, G. M., and J. G. Llaurado. Collagen and elastin content in canine arteries selected from functionally different vascular beds. Circ. Res. 19(2):394–399, 1966.

    PubMed  CAS  Google Scholar 

  9. Fung, Y. C. Wave Propagation in Blood Vessels, in Biodynamics Circulation. New York: Springer-Verlag, pp. 104–115, 1984.

    Google Scholar 

  10. Fung, Y., and P. Tong. Classical and Computational Solid Mechanics. World Scientific, 2001.

  11. Grassl, E. D., and J. C. Bischof. In vitro model systems for evaluation of smooth muscle cell response to cryoplasty. Cryobiology 50(2):162–173, 2005.

    Article  PubMed  Google Scholar 

  12. Han, B., J. D. Miller, and J. K. Jung. Freezing-induced fluid–matrix interaction in poroelastic material. J. Biomech. Eng. 131:021002, 2009.

    Article  PubMed  Google Scholar 

  13. Hong, J., et al. Hibernation induction trigger reduces hypoxic damage of swine skeletal muscle. Muscle Nerve 32(2):200–207, 2005.

    Article  PubMed  Google Scholar 

  14. Humphrey, J. D. Cardiovascular Solid Mechanics: Cells, Tissues, and Organs. Springer, 2002.

  15. Joye, J. D. The clinical application of cryoplasty for infrainguinal peripheral arterial disease. Tech. Vasc. Interv. Radiol. 8(4):160–164, 2005.

    Article  PubMed  Google Scholar 

  16. Kaushal, S., et al. Functional small-diameter neovessels created using endothelial progenitor cells expanded ex vivo. Nat. Med. 7:1035–1040, 2001.

    Article  PubMed  CAS  Google Scholar 

  17. Laird, J., et al. Cryoplasty for the treatment of femoropopliteal arterial disease: results of a prospective, multicenter registry. J. Vasc. Interv. Radiol. 16(8):1067–1073, 2005.

    PubMed  Google Scholar 

  18. Miles, C. A., T. V. Burjanadze, and A. J. Bailey. The kinetics of the thermal denaturation of collagen in unrestrained rat tail tendon determined by differential scanning calorimetry. J. Mol. Biol. 245(4):437–446, 1995.

    Article  PubMed  CAS  Google Scholar 

  19. Miles, C. A., et al. The increase in denaturation temperature following cross-linking of collagen is caused by dehydration of fibers. J. Mol. Biol. 346:551–556, 2005.

    Article  PubMed  CAS  Google Scholar 

  20. Moreaux, L., et al. Coherent scattering in multi-harmonic light microscopy. Biophys. J. 80(3):1568–1574, 2001.

    Article  PubMed  CAS  Google Scholar 

  21. Neidert, M. R., et al. Cryopreservation of collagen-based tissue equivalents. II. Effect of freezing in the presence of cryoprotective agents. Tissue Eng. 10(1/2):23–32, 2004.

    Article  PubMed  CAS  Google Scholar 

  22. Nerem, R. M. Tissue engineering a blood vessel substitute: the role of biomechanics. Yonsei Med. J. 41(6):735–739, 2000.

    PubMed  CAS  Google Scholar 

  23. Orihara, K., et al. Hyperthermia at 43 °C for 2 h inhibits the proliferation of vascular smooth muscle cells, but not endothelial cells. J. Mol. Cell. Cardiol. 34:1205–1215, 2002.

    Article  PubMed  CAS  Google Scholar 

  24. Rifkin, B. S., et al. Percutaneous balloon cryoplasty: a new therapy for rapidly recurrent anastomotic venous stenoses of hemodialysis grafts? Am. J. Kidney Dis. 45(2):e27–e32, 2005.

    Article  PubMed  Google Scholar 

  25. Rupp, C. C., et al. Cryosurgical changes in the porcine kidney: histologic analysis with thermal history correlation. Cryobiology 45:167–182, 2002.

    Article  PubMed  Google Scholar 

  26. Rupp, C. C., et al. Cryothermic and hyperthermic treatments of human leiomyomata and adjacent myometrium and their implications for laparoscopic surgery. Am. Assoc. Gynecol. Laparosc. 10(1):90–98, 2003.

    Article  Google Scholar 

  27. Shah, P. B., and C. M. Lilly. Interventional therapy for coronary artery disease. Am. J. Respir. Crit. Care Med. 166:791–796, 2002.

    Article  PubMed  Google Scholar 

  28. Shah, N. B., et al. Fourier transform infrared spectroscopy investigation of native tissue matrix modifications using a gamma irradiation process. Tissue Eng. Part C Methods 15(1):33–40, 2008.

    Article  CAS  Google Scholar 

  29. Sigg, D. C., and P. A. Iaizzo. Malignant hyperthermia phenotype. Hypotension induced by succinylcholine in susceptible swine. Anesthesiology 92(6):1777–1788, 2000.

    Article  PubMed  CAS  Google Scholar 

  30. Sokolis, D. P., H. Boudoulas, and P. E. Karayannacos. Assessment of the aortic stress-strain relation in uniaxial tension. J. Biomech. 35:1213–1223, 2002.

    Article  PubMed  Google Scholar 

  31. Song, Y. C., et al. Vitreous cryopreservation maintains the function of vascular grafts. Nat. Biotechnol. 18:296–299, 2000.

    Article  PubMed  CAS  Google Scholar 

  32. Strupler, M., et al. Second harmonic imaging and scoring of collagen in fibrotic tissues. Optics Express 15(7):4054–4065, 2007.

    Article  PubMed  CAS  Google Scholar 

  33. van Zandvoort, M., et al. Two-photon microscopy for imaging of the (atherosclerotic) vascular wall: a proof of concept study. J. Vasc. Res. 41(1):54–63, 2004.

    Article  PubMed  Google Scholar 

  34. Venkatasubramanian, R. T., et al. Effects of freezing and cryopreservation on the mechanical properties of arteries. Ann. Biomed. Eng. 34(5):823–832, 2006.

    Article  PubMed  Google Scholar 

  35. Vogt, P., et al. Technical details with the use of cryopreserved arterial allografts for aortic infection: influence on early and midterm mortality. J. Vasc. Surg. 35(1):80–86, 2002.

    PubMed  Google Scholar 

  36. Werkmeister, E., et al. Multiphoton microscopy for blood vessel imaging: new non-invasive tools (Spectral, SHG, FLIM). Clin. Hemorheol. Microcirc. 37(1):77–88, 2007.

    PubMed  CAS  Google Scholar 

  37. Williams, R., W. Zipfel, and W. Webb. Interpreting second-harmonic generation images of collagen I fibrils. Biophys. J. 88(2):1377–1386, 2005.

    Article  PubMed  CAS  Google Scholar 

  38. Wright, N. T., and J. D. Humphrey. Denaturation of collagen via heating: an irreversible rate process. Annu. Rev. Biomed. Eng. 4:109–128, 2002.

    Article  PubMed  CAS  Google Scholar 

  39. Wusteman, M., D. Pegg, and R. Warwick. The banking of arterial allografts in the United Kingdom. A technical and clinical review. Cell Tissue Bank. 1(4):295–301, 2000.

    Article  PubMed  Google Scholar 

  40. Zoumi, A., et al. Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy. Biophys. J. 87(4):2778–2786, 2004.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Dr. Schmechel for the histological analysis and Jerry Sedgewick for SHG microscopy. The authors would also like to thank the Experimental surgical services (ESS) and the Visible Heart laboratory at the University of Minnesota for access to fresh pig arteries. The authors would also like to thank Dr. Alptekin Aksan for the numerous discussions during the course of the study as well as Sue Clemmings for proofreading the manuscript. Funding from Boston Scientific, Institute for Engineering in Medicine at the University of Minnesota, and the Doctoral Dissertation Fellowship from the University of Minnesota Graduate School is gratefully acknowledged.

Author information

Author notes

  1. Wim F. Wolkers

    Present address: Institute for Multiphase Processes, Leibniz University of Hannover, Callinstr., 3630167, Hannover, Germany

Authors and Affiliations

  1. Department of Mechanical Engineering, University of Minnesota, 111 Church Street SE, Minneapolis, MN, 55455, USA

    Ramji T. Venkatasubramanian, Wim F. Wolkers & John C. Bischof

  2. Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, 55455, USA

    Mithun M. Shenoi, Victor H. Barocas & John C. Bischof

  3. Department of Urologic Surgery, University of Minnesota, Minneapolis, MN, 55455, USA

    John C. Bischof

  4. Department of Surgery, University of Minnesota, Minneapolis, MN, 55455, USA

    Charles L. Soule & Paul A. Iaizzo

  5. Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA

    Paul A. Iaizzo

  6. Boston Scientific Inc., Minneapolis, MN, USA

    Daniel Lafontaine

Authors
  1. Ramji T. Venkatasubramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wim F. Wolkers
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mithun M. Shenoi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Victor H. Barocas
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Daniel Lafontaine
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Charles L. Soule
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Paul A. Iaizzo
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. John C. Bischof
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to John C. Bischof.

Additional information

Associate Editor Julia E Babensee oversaw the review of this article.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Venkatasubramanian, R.T., Wolkers, W.F., Shenoi, M.M. et al. Freeze–Thaw Induced Biomechanical Changes in Arteries: Role of Collagen Matrix and Smooth Muscle Cells. Ann Biomed Eng 38, 694–706 (2010). https://doi.org/10.1007/s10439-010-9921-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10439-010-9921-9

Keywords

Search

Navigation