Skip to main content
SpringerLink
Log in

Tissue Oxygen Profiling Using Electron Paramagnetic Resonance Oximetry to Improve Wound Healing After Radiation

  • Original Paper
  • Published:
Applied Magnetic Resonance Aims and scope Submit manuscript

Abstract

The objective of this study is to describe the oxygen profile obtained by electron paramagnetic resonance (EPR) oximetry of tissue after radiation, surgery, and hyperbaric oxygen therapy (HBOT) and its relationship to wound healing in a rodent model. The study design is rodent model for wound healing. A rodent model for wound healing was used for oxygen measurements before and after various treatments. EPR measurements and biopsies of normal vs irradiated and flap vs non-flap tissues were taken at 1–3-week intervals for 12 weeks. Wound healing was evaluated by gross photos, histology, and immunostaining. Student’s t test and a linear mixed model were used to compare oxygen levels and gross healing with radiation exposure. A Proportional Odds model was also used to calculate odds ratio toward better wound-healing rate with radiation exposure. In the rodent model, at 1–3 weeks after irradiation, the mean tissue oxygen measurement was significantly lower in irradiated versus non-irradiated leg tissue. There was a significant difference in oxygenation between flap and non-flap tissue in an irradiated bed at 1 and 3 weeks after surgery. On gross evaluation, wound healing from z-plasty flap was significantly worse in irradiated tissue compared to non-irradiated tissue. A rodent model for wound healing showed that radiation resulted in decreased tissue oxygenation at 1–3 weeks after irradiation. Wound healing was compromised in irradiated tissue at earlier time points when tissue oxygenation was lower. Oxygen profiling with EPR oximetry can be used to identify timing of oxygen interventions to improve wound healing. Level of evidence is NA, animal studies.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Availability of Data and Materials

The data supporting the results reported in the article are available upon request.

References

  1. J.M. Arbeit, J.M. Brown, K.S.C. Chao et al., Hypoxia: importance in tumor biology, noninvasive measurement by imaging, and value of its measurement in the management of cancer therapy. Int. J. Radiat. Biol. 82, 699–757 (2006). https://doi.org/10.1080/09553000601002324

    Article  Google Scholar 

  2. H.W. Hopf, M.D. Rollins, Wounds: an overview of the role of oxygen. Antioxid. Redox Signal. 9, 1183–1192 (2007)

    Article  Google Scholar 

  3. C.K. Sen, Wound healing essentials: let there be oxygen. Wound Repair Regen. 17, 1–18 (2009). https://doi.org/10.1111/j.1524-475X.2008.00436.x

    Article  Google Scholar 

  4. S. Barrientos, O. Stojadinovic, M.S. Golinko et al., Growth factors and cytokines in wound healing. Wound Repair Regen. 16, 585–601 (2008). https://doi.org/10.1111/j.1524-475X.2008.00410.x

    Article  Google Scholar 

  5. G.M. Gordillo, S. Roy, S. Khanna et al., Topical oxygen therapy induces vascular endothelial growth factor expression and improves closure of clinically presented chronic wounds. Clin. Exp. Pharmacol. Physiol. 35, 957–964 (2008). https://doi.org/10.1111/j.1440-1681.2008.04934.x

    Article  Google Scholar 

  6. R.B. Fries, W.A. Wallace, S. Roy et al., Dermal excisional wound healing in pigs following treatment with topically applied pure oxygen. Mutat. Res. Fundam. Mol. Mech. Mutagen. 579, 172–181 (2005). https://doi.org/10.1016/j.mrfmmm.2005.02.023

    Article  Google Scholar 

  7. P. Kranke, M.H. Bennett, M. Martyn-St James et al., Hyperbaric Oxygen Therapy for Chronic Wounds (John Wiley and Sons Ltd., 2015)

    Book  Google Scholar 

  8. R. Choudhury, Hypoxia and hyperbaric oxygen therapy: a review. Int. J. Gen. Med. 11, 431–442 (2018). https://doi.org/10.2147/IJGM.S172460

    Article  Google Scholar 

  9. S.H. Ko, A.C. Nauta, S.D. Morrison et al., PHD-2 suppression in mesenchymal stromal cells enhances wound healing. Plast. Reconstr. Surg. 141, 55e–67e (2018). https://doi.org/10.1097/PRS.0000000000003959

    Article  Google Scholar 

  10. M.K. Tibbs, Wound healing following radiation therapy: a review. Radiother. Oncol. 42, 99–106 (1997)

    Article  Google Scholar 

  11. S. Delanian, J.L. Lefaix, Current management for late normal tissue injury: radiation-induced fibrosis and necrosis. Semin. Radiat. Oncol. 17, 99–107 (2007). https://doi.org/10.1016/j.semradonc.2006.11.006

    Article  Google Scholar 

  12. M.S. Anscher, L. Chen, Z. Rabbani et al., Recent progress in defining mechanisms and potential targets for prevention of normal tissue injury after radiation therapy. Int. J. Radiat. Oncol. Biol. Phys. 62, 255–259 (2005). https://doi.org/10.1016/j.ijrobp.2005.01.040

    Article  Google Scholar 

  13. H.B. Stone, C.N. Coleman, M.S. Anscher, W.H. McBride, Effects of radiation on normal tissue: consequences and mechanisms. Lancet Oncol. 4, 529–536 (2003). https://doi.org/10.1016/S1470-2045(03)01191-4

    Article  Google Scholar 

  14. H.B. Stone, W.H. McBride, C.N. Coleman, Modifying normal tissue damage postirradiation. Report of a workshop sponsored by the Radiation Research Program, National Cancer Institute. Radiat. Res. 157, 204–223 (2002). https://doi.org/10.1667/0033-7587(2002)157[0204:MNTDP]2.0.CO;2

    Article  ADS  Google Scholar 

  15. C.B. Westbury, A. Pearson, A. Nerurkar et al., Hypoxia can be detected in irradiated normal human tissue: a study using the hypoxic marker pimonidazole hydrochloride. Br. J. Radiol. 80, 934–938 (2007). https://doi.org/10.1259/bjr/25046649

    Article  Google Scholar 

  16. C.E. Fife, D.R. Smart, P.J. Sheffield et al., Transcutaneous oximetry in clinical practice: consensus statements from an expert panel based on evidence. Undersea Hyperb. Med. 36, 43–53 (2009)

    Google Scholar 

  17. A.B. Flood, V.A. Wood, H.M. Swartz, Using India ink as a sensor for oximetry: evidence of its safety as a medical device. Adv. Exp. Med. Biol. 977, 297–312 (2017). https://doi.org/10.1007/978-3-319-55231-6_40

    Article  Google Scholar 

  18. N. Khan, H. Hou, H.M. Swartz, P. Kuppusamy, Direct and Repeated Measurement of Heart and Brain Oxygenation Using in Vivo EPR Oximetry. In: Methods in Enzymology. Academic Press Inc. 564, 529–552 (2015)

  19. H. Hou, R. Dong, J.P. Lariviere et al., Synergistic combination of hyperoxygenation and radiotherapy by repeated assessments of tumor pO2 with EPR oximetry. J. Radiat. Res. 52, 568–574 (2011)

    Article  ADS  Google Scholar 

  20. M.A. Polacco, H. Hou, P. Kuppusamy, E.Y. Chen, Measuring flap oxygen using electron paramagnetic resonance oximetry. Laryngoscope 129, E415–E419 (2019). https://doi.org/10.1002/lary.28043

    Article  Google Scholar 

  21. C.M. Desmet, A. Lafosse, S. Vériter et al., Application of electron paramagnetic resonance (EPR) oximetry to monitor oxygen in wounds in diabetic models. PLoS ONE 10, 144914 (2015). https://doi.org/10.1371/journal.pone.0144914

    Article  Google Scholar 

  22. C.M. Desmet, G. Vandermeulen, C. Bouzin et al., EPR monitoring of wound oxygenation as a biomarker of response to gene therapy encoding hCAP-18/LL37 peptide. Magn. Reson. Med. 79, 3267–3273 (2018)

    Article  Google Scholar 

  23. M.M. Kmiec, H. Hou, M. Lakshmi Kuppusamy et al., Transcutaneous oxygen measurement in humans using a paramagnetic skin adhesive film. Magn. Reson. Med. 81, 781–794 (2019). https://doi.org/10.1002/mrm.27445

    Article  Google Scholar 

  24. B.F. Jordan, C. Baudelet, B. Gallez, Carbon-centered radicals as oxygen sensors for in vivo electron paramagnetic resonance: screening for an optimal probe among commercially available charcoals. Magn. Reson. Mater. Phys. Biol. Med. 7, 121–129 (1998)

    Article  Google Scholar 

  25. O.Y. Grinberg, H. Hou, S.A. Grinberg et al., pO2 and regional blood flow in a rabbit model of limb ischemia. Physiol. Meas. 25, 659–670 (2004). https://doi.org/10.1088/0967-3334/25/3/006

    Article  Google Scholar 

  26. G.A. Salam, J.P. Amin, T.J. Zuber, The basic Z-plasty. Am. Fam. Physician 67, 2329–2332 (2003)

    Google Scholar 

  27. M.S. Fazeli, M.G. Adel, A.H. Lebaschi, Comparison of outcomes in Z-plasty and delayed healing by secondary intention of the wound after excision of the sacral pilonidal sinus: results of a randomized, clinical trial. Dis. Colon Rectum 49, 1831–1836 (2006). https://doi.org/10.1007/s10350-006-0726-8

    Article  Google Scholar 

  28. N. Khan, B.B. Williams, H. Hou et al., Repetitive tissue pO2 measurements by electron paramagnetic resonance oximetry: current status and future potential for experimental and clinical studies. Antioxid. Redox Signal. 9, 1169–1182 (2007). https://doi.org/10.1089/ars.2007.1635

    Article  Google Scholar 

  29. N. Khan, S.P. Mupparaju, D. Mintzopoulos et al., Burn trauma in skeletal muscle results in oxidative stress as assessed by in vivo electron paramagnetic resonance. Mol. Med. Rep. 1, 813–819 (2008). https://doi.org/10.3892/mmr-00000033

    Article  Google Scholar 

  30. J.F. Dunn, H.M. Swartz, In vivo electron paramagnetic resonance oximetry with particulate materials. Methods 30, 159–166 (2003). https://doi.org/10.1016/S1046-2023(03)00077-X

    Article  Google Scholar 

  31. H.M. Swartz, H. Hou, N. Khan et al., Advances in probes and methods for clinical EPR oximetry. Adv. Exp. Med. Biol. 812, 73–79 (2014). https://doi.org/10.1007/978-1-4939-0620-8_10

    Article  Google Scholar 

  32. E. Demidenko, Mixed Models: Theory and Applications with R, 2nd edn. (Wiley, Hoboken, 2013)

    MATH  Google Scholar 

  33. D.M. Ansell, K.A. Holden, M.J. Hardman, Animal models of wound repair: Are they cutting it? Exp. Dermatol. 21, 581–585 (2012). https://doi.org/10.1111/j.1600-0625.2012.01540.x

    Article  Google Scholar 

  34. Z. Vujaskovic, M.S. Anscher, Q.F. Feng et al., Radiation-induced hypoxia may perpetuate late normal tissue injury. Int. J. Radiat. Oncol. Biol. Phys. 50, 851–855 (2001). https://doi.org/10.1016/S0360-3016(01)01593-0

    Article  Google Scholar 

  35. S. Auerswald, S. Schreml, R. Meier et al., Wound monitoring of pH and oxygen in patients after radiation therapy. Radiat. Oncol. (2019). https://doi.org/10.1186/s13014-019-1413-y

    Article  Google Scholar 

  36. M.S. Chin, B.B. Freniere, C.F. Bonney et al., Skin perfusion and oxygenation changes in radiation fibrosis. Plast. Reconstr. Surg. 131, 707–716 (2013). https://doi.org/10.1097/PRS.0b013e3182818b94

    Article  Google Scholar 

  37. J. Wright, Hyperbaric oxygen therapy for wound healing (2001). In: http://www.worldwidewounds.com/2001/april/Wright/HyperbaricOxygen.html. Accessed 14 Feb 2021

  38. S. Gehmert, S. Geis, P. Lamby et al., Evaluation of hyperbaric oxygen therapy for free flaps using planar optical oxygen sensors. Preliminary results. Clin. Hemorheol. Microcirc. 48, 75–79 (2011). https://doi.org/10.3233/CH-2011-1389

    Article  Google Scholar 

  39. J. Tlapák, P. Chmátal, J. Pejchal et al., The effect of hyperbaric oxygen therapy on acute wound healing in rabbits: an experimental stude and histopathological analysis. Mil Med Sci Lett 90, 2 (2021). https://doi.org/10.31482/mmsl.2021.001

    Article  Google Scholar 

  40. M.S. Anscher, B. Thrasher, Z. Rabbani et al., Antitransforming growth factor-β antibody 1D11 ameliorates normal tissue damage caused by high-dose radiation. Int. J. Radiat. Oncol. Biol. Phys. 65, 876–881 (2006). https://doi.org/10.1016/j.ijrobp.2006.02.051

    Article  Google Scholar 

  41. M.S. Anscher, B. Thrasher, L. Zgonjanin et al., Small molecular inhibitor of transforming growth factor-β protects against development of radiation-induced lung injury. Int. J. Radiat. Oncol. Biol. Phys. 71, 829–837 (2008). https://doi.org/10.1016/j.ijrobp.2008.02.046

    Article  Google Scholar 

  42. Y. Liu, K. Kudo, Y. Abe et al., Inhibition of transforming growth factor-beta, hypoxia-inducible factor-1alpha and vascular endothelial growth factor reduced late rectal injury induced by irradiation. J. Radiat. Res. 50, 233–239 (2009)

    Article  ADS  Google Scholar 

  43. J.W. Lee, J.P. Tutela, R.A. Zoumalan et al., Inhibition of Smad3 expression in radiation-induced fibrosis using a novel method for topical transcutaneous gene therapy. Arch. Otolaryngol. Head Neck Surg. 136, 714–719 (2010). https://doi.org/10.1001/archoto.2010.107

    Article  Google Scholar 

  44. C.A. Reddy, V. Somepalli, T. Golakoti et al., Mitochondrial-targeted curcuminoids: a strategy to enhance bioavailability and anticancer efficacy of curcumin. PLoS ONE 9, 1–11 (2014). https://doi.org/10.1371/journal.pone.0089351

    Article  Google Scholar 

  45. N. Charlier, N. Beghein, B. Gallez, Development and evaluation of biocompatible inks for the local measurement of oxygen using in vivo EPR. NMR Biomed. 17, 303–310 (2004). https://doi.org/10.1002/nbm.902

    Article  Google Scholar 

  46. G. Meenakshisundaram, E. Eteshola, R.P. Pandian et al., Fabrication and physical evaluation of a polymer-encapsulated paramagnetic probe for biomedical oximetry. Biomed. Microdevices 11, 773–782 (2009). https://doi.org/10.1007/s10544-009-9292-x

    Article  Google Scholar 

  47. P.E. Schaner, J.R. Pettus, A.B. Flood et al., OxyChip implantation and subsequent electron paramagnetic resonance oximetry in human tumors is safe and feasible: first experience in 24 patients. Front. Oncol. (2020). https://doi.org/10.3389/fonc.2020.572060

    Article  Google Scholar 

Download references

Acknowledgements

We acknowledge George Zhongzi Li for his consultation and assistance with the biostatistical analyses. We also acknowledge Lesley Jarvis, Jack Hoopes, Andrew Giustini, and Rendy Strawbridge for their assistance with the rat model and radiation treatments. We thank Thomas Matthews, Kevin Rychert, Huagang Hou, Nadeem Khan, Oleg Grinberg, and Hal Swartz for their assistance with EPR measurements and analyses as well as Jay Buckey, Donna Alvarenga, Hermine Wallach, and Nancy Yazinski for their help with the hyperbaric oxygen treatments.

Funding

These studies were supported by The Triological Society Career Development Award and pilot award from the Norris Cotton Cancer Center’s American Cancer Society Institutional Research Grant (IRG-82-003-27).

Author information

Authors and Affiliations

  1. Department of Surgery, Section of Otolaryngology, Dartmouth-Hitchcock Medical Center, 1 Medical Center Drive, Lebanon, NH, 03756, USA

    Eunice Y. Chen

  2. Geisel School of Medicine at Dartmouth, Hanover, NH, USA

    Eunice Y. Chen & Benjamin B. Williams

  3. Thayer School of Engineering, Dartmouth College, Hanover, NH, USA

    Sassan Hodge

  4. Section of Radiation Oncology, Department of Medicine, Geisel School of Medicine, Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA

    Benjamin B. Williams

Authors
  1. Eunice Y. Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sassan Hodge
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Benjamin B. Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

EYC and BBW contributed to the study conception and design. Material preparation and data collection were performed by EYC and SH. Analyses were performed by all authors. The first draft of the manuscript was written by EYC and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Eunice Y. Chen.

Ethics declarations

Conflict of interest

The authors do not have any conflicts of interest or competing interests to disclose.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

723_2021_1367_MOESM1_ESM.pdf

Supplementary Timeline 1. Schedule of char implantations, EPR measurements, treatments, and procedures in rat model for studies excluding (A) and including (B) hyperbaric oxygen therapy. Key: EPR, electron paramagnetic resonance oximetry; XRT, radiation therapy; HBOT, hyperbaric oxygen therapy; W, week (PDF 256 KB)

Supplementary Figure 1 Schematic drawing of rat model for wound healing (PDF 304 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, E.Y., Hodge, S. & Williams, B.B. Tissue Oxygen Profiling Using Electron Paramagnetic Resonance Oximetry to Improve Wound Healing After Radiation. Appl Magn Reson 52, 1489–1507 (2021). https://doi.org/10.1007/s00723-021-01367-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00723-021-01367-6

Search

Navigation