|Year : 2016 | Volume
| Issue : 3 | Page : 138-146
Temporal variation in the response of tumors to hyperoxia with breathing carbogen and oxygen
Hua-gang Hou M.D. 1, Nadeem Khan1, Gai-xin Du2, Sassan Hodge3, Harold M Swartz1
1 EPR Center for Viable Systems, Geisel School of Medicine at Dartmouth, Lebanon, NH; Norris Cotton Cancer Center, Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA
2 EPR Center for Viable Systems, Geisel School of Medicine at Dartmouth, Lebanon, NH, USA
3 Department of Genetics, Geisel School of Medicine at Dartmouth, Hanover, NH, USA
|Date of Web Publication||14-Oct-2016|
EPR Center for Viable Systems, Geisel School of Medicine at Dartmouth, Lebanon, NH; Norris Cotton Cancer Center, Dartmouth-Hitchcock Medical Center, Lebanon, NH
Source of Support: This work was supported by NIH grant P01EB2180 and Hitchcock Foundation Program Project Grant and the Prouty Grants from the NCCC at Dartmouth-Hitchcock Medical Center., Conflict of Interest: None
The effect of hyperoxygenation with carbogen (95% O 2 + 5% CO 2 ) and 100% oxygen inhalation on partial pressure of oxygen (pO 2 ) of radiation-induced fibrosarcoma (RIF-1) tumor was investigated. RIF-1 tumors were innoculated in C3H mice, and aggregates of oximetry probe, lithium phthalocyanine (LiPc), was implanted in each tumor. A baseline tumor pO 2 was measured by electron paramagnetic resonance (EPR) oximetry for 20 minutes in anesthetized mice breathing 30% O 2 and then the gas was switched to carbogen or 100 % oxygen for 60 minutes. These experiments were repeated for 10 days. RIF-1 tumors were hypoxic with a baseline tissue pO 2 of 6.2-8.3 mmHg in mice breathing 30% O 2 . Carbogen and 100% oxygen significantly increased tumor pO 2 on days 1 to 5, with a maximal increase at approximately 32-45 minutes on each day. However, the extent of increase in pO 2 from the baseline declined significantly on day 5 and day 10. The results provide quantitative information on the effect of hyperoxic gas inhalation on tumor pO 2 over the course of 10 days. EPR oximetry can be effectively used to repeatedly monitor tumor pO 2 and test hyperoxic methods for potential clinical applications.
Keywords: carbogen; electron paramagnetic resonance; hyperoxia; multi-site electron paramagnetic resonance oximetry; oximetry; oxygen; partial pressure of oxygen; radiation-induced fibrosarcoma
|How to cite this article:|
Hou Hg, Khan N, Du Gx, Hodge S, Swartz HM. Temporal variation in the response of tumors to hyperoxia with breathing carbogen and oxygen. Med Gas Res 2016;6:138-46
|How to cite this URL:|
Hou Hg, Khan N, Du Gx, Hodge S, Swartz HM. Temporal variation in the response of tumors to hyperoxia with breathing carbogen and oxygen. Med Gas Res [serial online] 2016 [cited 2019 Nov 20];6:138-46. Available from: http://www.medgasres.com/text.asp?2016/6/3/138/191359
| Introduction|| |
The efficacy of any treatment modality is largely influenced by the tumor microenvironment. One of the most important factors that influence the treatment outcome is the existence of hypoxia in solid tumors. It has been shown that hypoxia reduce the efficacy of radiation treatment, surgery and chemotherapy (Fyles et al., 1998; Knocke et al., 1999; Kaanders et al., 2002c; Nordsmark et al., 2005). A direct correlation between the oxygen levels in the tumors and treatment outcome has been reported (Nordsmark and Overgaard, 2000; Moeller et al., 2007; Kappler et al., 2008; Lovey et al., 2008; Rockwell et al., 2009). Consequently, an increase in the oxygen levels can improve the treatment outcome of tumors (Rudat et al., 2000; Robinson et al., 2001; Schuuring et al., 2002, 2006). Carbogen (95% O2 + 5% CO2) and 100% oxygen are the most convenient methods as they involve direct inhalation and do not inflict any toxicity (Overgaard and Horsman, 1996; Kulkarni et al., 2007). Breathing carbogen in combination with nicotinamide administration was used to improve tumor response to accelerated radiotherapy in head-and-neck tumors (Saunders et al., 1997; Kaanders et al., 1998, 2002b; Bernier et al., 2000; Janssens et al., 2012). However, only a poor to modest improvement in the treatment outcome was observed in the clinical trials (Bernier et al., 2000; Kaanders et al., 2002a). This is likely due to a lack of knowledge about the oxygen levels in the tumors during treatments with hyperoxic approaches.
Techniques that can provide direct and repeated measurements of tumor partial pressure of oxygen (pO2) will be extremely useful to detect therapeutic window during which the tumors response to hyperoxic strategies and then schedule radiotherapy to improve treatment outcome. The oxygen electrode technique has been considered the gold standard for assessing tumor oxygenation in the clinic (Evans and Koch, 2003; Milosevic et al., 2004; Williams et al., 2005). This approach provides a direct and rapid measurement of tissue pO2 , but use of the electrodes has a number of limitations. These include their applicability only to easily accessible tumors and the failure to distinguish necrosis from hypoxic viable tissue. Furthermore, oxygen electrodes cannot provide repeated measurements of temporal changes in pO2 that is vital in determining the efficacy of anticancer treatments (Evans and Koch, 2003; Milosevic et al., 2004; Williams et al., 2005). Methods based on nuclear magnetic resonance (NMR) principles also have been developed such as19 F-NMR spectroscopy (Hunjan et al., 2001), blood oxygen level dependent (BOLD) imaging (Baudelet and Gallez, 2002), and Overhauser methods (Krishna et al., 2002). While these methods have provided useful data and have the benefit of widely available instrumentation, each has a limitation for in vivo applications.19 F-NMR spectroscopy requires the injection of the probe directly into the tumor and therefore has limited ability to make repeated measurements during therapy. BOLD imaging does not require any injection, but the information that it provides, the total amount of deoxyhemoglobin in the blood, cannot be related directly to the oxygen levels in the tumor. Overhauser method also relies on the repeated injection of paramagnetic material and has limited sensitivity. In vivo electron paramagnetic resonance (EPR) oximetry has the ability to provide repeated measurements of tumor pO2 to test and optimize strategies designed for hyperoxygenation and demonstrate therapeutic benefits (Hou et al., 2004, 2007, 2009, 2010, 2011, 2013; Khan et al., 2009, 2010). We have systematically investigated the effect of breathing carbogen and 100% oxygen on the pO2 in subcutaneous radiation-induced fibrosarcoma (RIF-1) tumors in the experiments repeated for up to 10 days. The maximum increase in tumor pO2 , time to reach maximum pO2 (Tmax) and the percentage of tumors that showed a significant increase in pO2 were used to evaluate the effectiveness of breathing carbogen and 100% O2 inhalation for hyperoxia.
| Materials and Methods|| |
Animals and tumor models
All animal procedures were in accordance with the NIH Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of Dartmouth Medical School (Geisel School of Medicine). The RIF-1 cells were a gift from Dr. J. B. Mitchell's laboratory at the National Cancer Institute. This well-established subcutaneous tumor model has been used for several studies in our laboratory (Hou et al., 2004, 2007, 2009, 2010, 2011). The cells were cultured in vitro in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), glutamine and antibiotics. The procedure for tumor inoculation has been described previously (Hou et al., 2004, 2007, 2009, 2010, 2011). Briefly, female C3H/HEJ mice (18-20 g, Charles River Laboratories, Wilmington, MA, USA) were anesthetized (1.2% isoflurane, 30% O2), and a suspension of 2-3 × 105 RIF-1 cells in 100 μL was injected subcutaneously into the left posterior flank. The tumors reached a size of 150-200 mm3 in approximately 12-14 days post cell inoculation.
Implantation of the lithium phthalocyanine (LiPc) oximetry probe
The LiPc crystals were synthesized in our laboratory and the physico-chemical properties of LiPc crystals have been described previously (Liu et al., 1993; Swartz and Walczak, 1998). The LiPc crystals have a single sharp EPR line whose width is highly sensitive to pO2 . The EPR spectra reflect the average pO2 on the surface of each LiPc aggregate (30-50 μg with an approximate volume of 0.09-0.15 mm3) injected in the tumors. The procedure for LiPc injection has been described earlier (Hou et al., 2009, 2011, 2013). Briefly, the mice were anesthetized (1.5% isoflurane with 30% O2), and two aggregates of the LiPc crystals were implanted at a depth of 2 mm and at a distance of 4 mm into each tumor using 25-gauge needles. A baseline tumor pO2 were measured 24 hours after LiPc implantation for 30 minutes in anesthetized mice (1.5% isoflurane with 30% O2) by using multi-site EPR oximetry described below. The tissue pO2 measured from two LiPc implants in each tumor were pooled to determine average pO2 on each day (Hou et al., 2009, 2011, 2013).
The mice were randomly assigned into six groups which were allowed to breath normobaric carbogen (group A) or 100% O2 (group B) multiple times on days 1-5 and day 10 or breathing carbogen once on day 1 (group C), day 3 (group D), day 5 (group E) and day 10 (group F) for 60 minutes after 30 minutes of baseline pO2 measurements in anesthetized mice breathing 30% O2 ([Table 1]).
Multi-site EPR oximetry
Assessment of tissue pO2 at 2-4 sites simultaneously and repeatedly by EPR oximetry has been used in animal models (Hou et al., 2009, 2011, 2013). The pO2 measurements were performed on an L-band (1,200 MHz) EPR spectrometer designed for in vivo experiments (Swartz and Walczak, 1998). A set of coils capable of generating a magnetic field gradient in the Z-direction with a magnitude up to 3.0 G/cm was used to separate the EPR spectra of LiPc implants in each tumor (Smirnov et al., 1993). The spectrometer parameters were: incident microwave power, 0.8-8.0 mW; magnetic field center, 425 gauss; scan range, 2 gauss; modulation frequency, 24 kHz; modulation amplitude was one-third of the EPR line width with scan time of 10 seconds. The EPR line-widths were converted to pO2 using a calibration determined for each batch of LiPc crystals (Liu et al., 1993; Hou et al., 2009, 2013).
Physiological control and histological analysis
During EPR oximetry measurements, the body temperature of the animals was monitored using a rectal probe and animals were maintained at 37.0 ± 0.5°C using a thermostatically controlled heated pad and a flow of warm air.
At the end of experiments, the animals were euthanized, tumors removed, fixed, and sectioned. Microscopic examination (hematoxylin-eosin staining) of the tissue around the implanted LiPc deposits was performed to assess the tumor micro-environment.
A paired t-test was used to determine the statistical significance of the changes in pO2 and tumor volume within the group and an unpaired t-test was used to determine the significance between groups. The paired comparison reduces the effects of animal to animal heterogeneity and eliminates differences of the baseline pO2 . The chi-square test was used to compare the percentage (%) of tumors with an increase in pO2 of more than 50% from the baseline during first 20 minutes of breathing carbogen or oxygen compared to day 1. The multilevel linear mixed effects model was used to analyze the longitudinal pO2 data in the log scale (Demidenko and Stukel, 2002; Demidenko, 2004). An exponential quadratic function of time was used to determine maximum pO2 (pO2max) and Tmax during carbogen and 100% oxygen inhalation. Each curve was analyzed accounting for two sources of variation: inter- and intra-mouse variations. Calculations were done by the statistical package S-Plus version 6.2 (Insight Inc., Seattle, WA, USA). All data of mean baseline and changes in pO2 including pO2max and tumor volume are expressed as the mean ± SEM. The tests were two-sided, and a change with P < 0.05 was considered significant. n is the number of the implant, N is the number of animals in each group.
| Results|| |
Effect of breathing carbogen or 100% O2 on tumor pO2 and Tmax
[Figure 1]A and [Figure 1]B show absolute and mean baseline tumor pO2 from individual LiPc implants on days 1 to 5 and day 10 in the groups A and B, respectively. The mean baseline pO2 in groups A and B were 6.2 ± 0.9 mmHg and 8.3 ± 1.2 mmHg respectively on day1 ([Table 2]). The mean baseline tumor volumes on day 1 were 146 ± 10 mm3 in group A and 167 ± 8 mm3 in group B. No significant changes in the tumor pO2 were observed by day 10; however the tumor volume increased significantly to 931 ± 64 mm3 and 1,035 ± 65 mm3 in group A and B, respectively ([Table 3]). There were no apparent effects of tumor volume on pO2 (group A: r = 0.24; group B: r = 0.14).
|Table 2: Baseline pO2 (mmHg), maximum pO2 (mmHg) and time to reach to maximum pO2 (minutes) in different groups on day 1 to day 5 and day 10|
Click here to view
The mean baseline pO2 and an increase in pO2 averaged over 20 minutes during 60 minutes of carbogen and 100% O2 inhalations are shown in [Figure 2]A and [Figure 2]B, respectively. The mean tumor pO2 during carbogen and 100% O2 inhalation on days 1-5 were significantly greater than the baseline pO2 (P < 0.05 or P < 0.01). The exponential quadratic function of time indicated a maximal increase in pO2 at 34.1 ± 2.1 to 41.8 ± 2.6 minutes on day 1 and 35.6 ± 4.6 to 43.0 ± 2.5 minutes on day 5. However, no significant increase in tumor pO2 on breathing carbogen or 100% O2 was evident on day 10 ([Figure 3], [Table 2]).
|Figure 3: Time course of tumor pO2 measured in individual mice prior to and during (A) carbogen and (B) 100% O2 inhalation.|
Each line is pO2 over time from an implant (2 implants/animal) from all animals (Ngroup A = 9; Ngroup B= 5), therefore, 16-18 lines for group A and 8-10 lines for group B per day. The bold line shows the average response of tumor pO2 and the star * and the horizontal line shows the averaged maximum pO2 (pO2 max) and averaged time to reach to maximum pO2 (Tmax) for all animals in the group using an exponential quadratic function. pO2: Partial pressure of oxygen; min: minute(s)
Click here to view
The tumors were categorized based on their increase in pO2 of more than 50% from the baseline within 20 minutes of breathing carbogen or 100% oxygen on each day, [Figure 4]A and [Figure 4]B, respectively. These results indicate a significant decrease in the percentage of tumors that responded to carbogen or 100% O2 on day 5 (P < 0.05) and day 10 (P < 0.01) compared to day 1.
|Figure 4: The percentage change in the number of tumors with pO2 greater than 50% of the baseline (black column) and less than 50% of the baseline (white column) within the first 20 minutes of carbogen (A) and 100% O2 (B) inhalation on each day.|
Note: *P < 0.05, **P < 0.01, vs. day 1. pO2: Partial pressure of oxygen
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Effect of single administration of carbogen on tumor pO2 and Tmax
The absolute baseline tumor pO2 from individual LiPc aggregates and the mean tumor pO2 on day 1 (group C), day 3 (group D), day 5 (group E) or day 10 (group F) are shown in [Figure 5]A. The mean baseline pO2 were 6.6 ± 0.8 mmHg, 6.9 ± 0.6 mmHg, 5.4 ± 0.4 mmHg, 6.0 ± 0.7 mmHg in groups C, D, E and F, respectively ([Table 2]). The mean tumor volumes as shown in [Table 3] were not significantly different than groups A and B on the corresponding days. The increase in the mean tumor pO2 during carbogen inhalation was significantly higher than the baseline pO2 (P < 0.05; [Figure 5]B). Maximal increase in tumor pO2 was observed at 41.2 ± 3.4 minutes on day 1, 37.7 ± 2.0 minutes on day 3 and 36.3 ± 2.8 minutes on day 5. However, no significant maximum increase in tumor pO2 was observed on day 10 ([Figure 5]C, [Table 2]).
The percentage of tumors with an increase in pO2 of more than 50% from the baseline within 20 minutes of breathing carbogen on days 1, 3, 5 and day 10 are shown in [Figure 5]D. The results indicate a significant decline in the tumors that responded to carbogen on day 5 (P < 0.05) and day 10 (P < 0.01) compared to day 1.
Effect of breathing carbogen or 100% O2 on morphologic examination of the tumor
Gross and microscopic examination (hematoxylin-eosin staining) of the tissue confirmed that the LiPc aggregates were in the interstitial compartment of the tumor tissue with no evidence of edema or infiltration of inflammatory cells, although accumulation of blood cells and necrotic cells around the LiPc deposits was observed in some samples on day 5 (group E) and day 10 (group F) ([Figure 6]).
|Figure 6: Effect of breathing carbogen on morphologic examination of the RIF-1 tumors (hematoxylin-eosin staining).|
Note: The LiPc crystals (black color) are indicated by short black arrows, necrotic tissues are indicated by long white arrows and the blood cell accumulation is indicated by short white color dash line arrows. M4, M6, M7 and M8 represent the mouse numbers. The thickness of each section is 5 ?m. Magnification: 1× (upper panel) and 10× (lower panel).
Click here to view
| Discussion|| |
The results reported here is the continuation of our systematic study to characterize the changes in tumor pO2 during the inhalation of hyperoxic gases by EPR oximetry. We have previously reported the effect of breathing carbogen on the RIF-1, F98, C6 and 9L tumors pO2 with the goal to improve oxygen levels for radiosensitization (Khan et al., 2009, 2010; Hou et al., 2011, 2012). The current results indicate that the RIF-1 tumors are hypoxic with a pO2 of < 10 mmHg, consistent with earlier reports (Hasegawa et al., 1987; Hou et al., 2004, 2007, 2009, 2010, 2011, 2013). Kavanagh et al. (1996) reported a mean of 4.5 mmHg, when the diameter of RIF-1 tumor reached 11 ± 0.5 mmHg. With EPR oximetry, Bratasz et al. (2007) reported an average pO2 of 4.9 mmHg in subcutaneous RIF-1 tumors of 113 mm3 volume.
The temporal changes in tumor pO2 with carbogen inhalation has varied in studies using different tumor types. Bussink et al. (2000) reported a rapid increase in the tumor pO2 , with the mean Tmax at approximately 3.9 (0.8-14) minutes and 3.6 (0.7-16) minutes in E102 and E106 glioblastoma xenografts, respectively. Gu et al. (2003) reported a significant increase in subcutaneous mammary adenocarcinoma tumors pO2 within 8 minutes of carbogen breathing which gradually increased over the next 12 minutes. Thews et al. (2002) observed a slow increase in the pO2 of subcutaneous rat DS-sarcomas over 15 minutes during carbogen challenge. We have observed a significant increase in the intracerebral F98 tumor pO2 within 15 minutes of carbogen inhalation (Hou et al., 2012). On the other hand, a significant increase in the RIF-1 tumor pO2 , with a Tmax at 22.9-31.2 minutes on days 1-5 post carbogen inhalation was observed (Hou et al., 2011). In the present study, carbogen inhalation significantly increased tumor pO2 , with a maximum increase at 32-44 minutes on days 1-5. These results indicate that the time to achieve a maximal increase in tumor pO2 with carbogen inhalation depends on the tumor type, tumor location, and possibly tumor size. A significant decline in tumor pO2 during repeated breathing of 100% oxygen inhalation over days was also observed. Similar results has been observed previously in RIF-1 (Hou et al., 2011) and also 9L, C6 tumor models (Khan et al., 2009). This is likely due to a combination of several factors, such as an increase in interstitial pressure, compromised tumor vasculature with increase in tumor size and increase in necrotic areas in the tumor over days (Khan et al., 2009).
Several investigators have used percentage pO2 values ≤ 5 mmHg as a measure of a clinically significant hypoxic fraction. Fyles et al. (1998) presented the hypoxic proportion, defined as the percentage of pO2 values less than the median value of 5 mmHg in patients with cervix cancer and found grouping of patients above and below the percentage of pO2 readings of < 5 mmHg of 50% (closely approximating the median value of 52%) resulted in a 2.9-fold greater risk of failure for hypoxic tumors. A group of patients with bulky hypoxic tumors appears to have a substantially higher risk of tumor recurrence or death tumors with > 50% change in pO2 from pretreatment had a poor outcome. We have observed a significant decline in the percentage of tumors with an increase in pO2 of more than 50% from the baseline on day 5 and day 10 compared to day 1 during multiple or single administration of carbogen. These results indicate that the response to carbogen is not consistent over days and therefore likely to compromise the efficacy of fractionated radiotherapy, which is usually administered for 4-6 weeks. On the other hand, fractionated radiotherapy can also alter the response of tumors to carbogen due to cell kill and effect on the tumor vasculature (Song et al., 1987; Yaromina et al., 2011; Multhoff and Vaupel, 2012). This will be the focus of our future study in the quest to effectively use carbogen to oxygenate and radiosensitize the tumors. The histological results indicate some accumulation of necrotic cells around the LiPc aggregates in some samples on day 5 (group E) and day 10 (group F). These were not caused by the LiPc implants, rather reflects the normal histological pattern of the tumor. These results are consistent with our previous report on the histological appearance of solid tumors (Hou et al., 2013).
In conclusion, these results provide quantitative information on the effect of carbogen and 100% oxygen inhalation in enhancing tumor oxygenation over the course of 10 days. Since the time to achieve a significant increase in tumor pO2 is likely to vary with the tumor type, size, and site, EPR oximetry can be used to repeatedly monitor tumor pO2 to test hyperoxic interventions and confirm tumor oxygenation.
| References|| |
Baudelet C, Gallez B (2002) How does blood oxygen level-dependent (BOLD) contrast correlate with oxygen partial pressure (pO 2
) inside tumors? Magn Reson Med 48:980-986.
Bernier J, Denekamp J, Rojas A, Minatel E, Horiot J, Hamers H, Antognoni P, Dahl O, Richaud P, van Glabbeke M, Pierart M (2000) ARCON: accelerated radiotherapy with carbogen and nicotinamide in head and neck squamous cell carcinomas. The experience of the Co-operative group of radiotherapy of the european organization for research and treatment of cancer (EORTC). Radiother Oncol 55:111-119.
Bratasz A, Pandian RP, Deng Y, Petryakov S, Grecula JC, Gupta N, Kuppusamy P (2007) In vivo imaging of changes in tumor oxygenation during growth and after treatment. Magn Reson Med 57:950-959.
Bussink J, Kaanders JH, Strik AM, van der Kogel AJ (2000) Effects of nicotinamide and carbogen on oxygenation in human tumor xenografts measured with luminescense based fiber-optic probes. Radiother Oncol 57:21-30.
Demidenko E (2004) Mixed Models: Theory and Applications. New York: Wiley.
Demidenko E, Stukel T (2002) Efficient estimation of general mixed effects models. J Stat Plan Infer 104:197-219.
Evans SM, Koch CJ (2003) Prognostic significance of tumor oxygenation in humans. Cancer Lett 195:1-16.
Fyles AW, Milosevic M, Wong R, Kavanagh MC, Pintilie M, Sun A, Chapman W, Levin W, Manchul L, Keane TJ, Hill RP (1998) Oxygenation predicts radiation response and survival in patients with cervix cancer. Radiother Oncol 48:149-156.
Gu Y, Bourke VA, Kim JG, Constantinescu A, Mason RP, Liu H (2003) Dynamic response of breast tumor oxygenation to hyperoxic respiratory challenge monitored with three oxygen-sensitive parameters. Appl Opt 42:2960-2967.
Hasegawa T, Rhee JG, Levitt SH, Song CW (1987) Increase in tumor pO2 by perfluorochemicals and carbogen. Int J Radiat Oncol Biol Phys 13:569-574.
Hou H, Lariviere JP, Demidenko E, Gladstone D, Swartz H, Khan N (2009) Repeated tumor pO(2) measurements by multi-site EPR oximetry as a prognostic marker for enhanced therapeutic efficacy of fractionated radiotherapy. Radiother Oncol 91:126-131.
Hou H, Abramovic Z, Lariviere JP, Sentjurc M, Swartz H, Khan N (2010) Effect of a topical vasodilator on tumor hypoxia and tumor oxygen guided radiotherapy using EPR oximetry. Radiat Res 173:651-658.
Hou H, Dong R, Lariviere JP, Mupparaju SP, Swartz HM, Khan N (2011) Synergistic combination of hyperoxygenation and radiotherapy by repeated assessments of tumor pO2 with EPR oximetry. J Radiat Res 52:568-574.
Hou H, Mupparaju SP, Lariviere JP, Hodge S, Gui J, Swartz HM, Khan N (2013) Assessment of the changes in 9L and C6 glioma pO2 by EPR oximetry as a prognostic indicator of differential response to radiotherapy. Radiat Res 179:343-351.
Hou H, Khan N, Grinberg OY, Yu H, Grinberg SA, Lu S, Demidenko E, Steffen RP, Swartz HM (2007) The effects of Efaproxyn (efaproxiral) on subcutaneous RIF-1 tumor oxygenation and enhancement of radiotherapy-mediated inhibition of tumor growth in mice. Radiat Res 168:218-225.
Hou H, Dong R, Li H, Williams B, Lariviere JP, Hekmatyar SK, Kauppinen RA, Khan N, Swartz H (2012) Dynamic changes in oxygenation of intracranial tumor and contralateral brain during tumor growth and carbogen breathing: a multisite EPR oximetry with implantable resonators. J Magn Reson 214:22-28.
Hou H, Khan N, O'Hara JA, Grinberg OY, Dunn JF, Abajian MA, Wilmot CM, Makki M, Demidenko E, Lu S, Steffen RP, Swartz HM (2004) Effect of RSR13, an allosteric hemoglobin modifier, on oxygenation in murine tumors: an in vivo electron paramagnetic resonance oximetry and bold MRI study. Int J Radiat Oncol Biol Phys 59:834-843.
Hunjan S, Zhao D, Constantinescu A, Hahn EW, Antich PP, Mason RP (2001) Tumor oximetry: demonstration of an enhanced dynamic mapping procedure using fluorine-19 echo planar magnetic resonance imaging in the Dunning prostate R3327-AT1 rat tumor. Int J Radiat Oncol Biol Phys 49:1097-1108.
Janssens GO, Rademakers SE, Terhaard CH, Doornaert PA, Bijl HP, van den Ende P, Chin A, Marres HA, de Bree R, van der Kogel AJ, Hoogsteen IJ, Bussink J, Span PN, Kaanders JH (2012) Accelerated radiotherapy with carbogen and nicotinamide for laryngeal cancer: results of a phase III randomized trial. J Clin Oncol 30:1777-1783.
Kaanders JH, Bussink J, van der Kogel AJ (2002a) ARCON: a novel biology-based approach in radiotherapy. Lancet Oncol 3:728-737.
Kaanders JH, Pop LA, Marres HA, Liefers J, van den Hoogen FJ, van Daal WA, van der Kogel AJ (1998) Accelerated radiotherapy with carbogen and nicotinamide (ARCON) for laryngeal cancer. Radiother Oncol 48:115-122.
Kaanders JH, Pop LA, Marres HA, Bruaset I, van den Hoogen FJ, Merkx MA, van der Kogel AJ (2002b) ARCON: experience in 215 patients with advanced head-and-neck cancer. Int J Radiat Oncol Biol Phys 52:769-778.
Kaanders JH, Wijffels KI, Marres HA, Ljungkvist AS, Pop LA, van den Hoogen FJ, de Wilde PC, Bussink J, Raleigh JA, van der Kogel AJ (2002c) Pimonidazole binding and tumor vascularity predict for treatment outcome in head and neck cancer. Cancer Res 62:7066-7074.
Kappler M, Taubert H, Holzhausen HJ, Reddemann R, Rot S, Becker A, Kuhnt T, Dellas K, Dunst J, Vordermark D, Hansgen G, Bache M (2008) Immunohistochemical detection of HIF-1alpha and CAIX in advanced head-and-neck cancer. Prognostic role and correlation with tumor markers and tumor oxygenation parameters. Strahlenther Onkol 184:393-399.
Kavanagh MC, Sun A, Hu Q, Hill RP (1996) Comparing techniques of measuring tumor hypoxia in different murine tumors: Eppendorf pO 2
Histograph, [3H]misonidazole binding and paired survival assay. Radiat Res 145:491-500.
Khan N, Li H, Hou H, Lariviere JP, Gladstone DJ, Demidenko E, Swartz HM (2009) Tissue pO 2
of orthotopic 9L and C6 gliomas and tumor-specific response to radiotherapy and hyperoxygenation. Int J Radiat Oncol Biol Phys 73:878-885.
Khan N, Mupparaju S, Hekmatyar SK, Hou H, Lariviere JP, Demidenko E, Gladstone DJ, Kauppinen RA, Swartz HM (2010) Effect of hyperoxygenation on tissue pO 2
and its effect on radiotherapeutic efficacy of orthotopic F98 gliomas. Int J Radiat Oncol Biol Phys 78:1193-1200.
Knocke TH, Weitmann HD, Feldmann HJ, Selzer E, Potter R (1999) Intratumoral pO 2
-measurements as predictive assay in the treatment of carcinoma of the uterine cervix. Radiother Oncol 53:99-104.
Krishna MC, English S, Yamada K, Yoo J, Murugesan R, Devasahayam N, Cook JA, Golman K, Ardenkjaer-Larsen JH, Subramanian S, Mitchell JB (2002) Overhauser enhanced magnetic resonance imaging for tumor oximetry: coregistration of tumor anatomy and tissue oxygen concentration. Proc Natl Acad Sci U S A 99:2216-2221.
Kulkarni AC, Kuppusamy P, Parinandi N (2007) Oxygen, the lead actor in the pathophysiologic drama: enactment of the trinity of normoxia, hypoxia, and hyperoxia in disease and therapy. Antioxid Redox Signal 9:1717-1730.
Liu KJ, Gast P, Moussavi M, Norby SW, Vahidi N, Walczak T, Wu M, Swartz HM (1993) Lithium phthalocyanine: a probe for electron paramagnetic resonance oximetry in viable biological systems. Proc Natl Acad Sci U S A 90:5438-5442.
Lovey J, Bereczky B, Gilly R, Kenessey I, Raso E, Simon E, Dobos J, Vago A, Kasler M, Dome B, Timar J, Tovari J (2008) Recombinant human erythropoietin alpha improves the efficacy of radiotherapy of a human tumor xenograft, affecting tumor cells and microvessels. Strahlenther Onkol 184:1-7.
Milosevic M, Fyles A, Hedley D, Hill R (2004) The human tumor microenvironment: invasive (needle) measurement of oxygen and interstitial fluid pressure. Semin Radiat Oncol 14:249-258.
Moeller BJ, Richardson RA, Dewhirst MW (2007) Hypoxia and radiotherapy: opportunities for improved outcomes in cancer treatment. Cancer Metastasis Rev 26:241-248.
Multhoff G, Vaupel P (2012) Radiation-induced changes in microcirculation and interstitial fluid pressure affecting the delivery of macromolecules and nanotherapeutics to tumors. Front Oncol 2:165.
Nordsmark M, Overgaard J (2000) A confirmatory prognostic study on oxygenation status and loco-regional control in advanced head and neck squamous cell carcinoma treated by radiation therapy. Radiother Oncol 57:39-43.
Nordsmark M, Bentzen SM, Rudat V, Brizel D, Lartigau E, Stadler P, Becker A, Adam M, Molls M, Dunst J, Terris DJ, Overgaard J (2005) Prognostic value of tumor oxygenation in 397 head and neck tumors after primary radiation therapy. An international multi-center study. Radiother Oncol 77:18-24.
Overgaard J, Horsman MR (1996) Modification of hypoxia-induced radioresistance in tumors by the use of oxygen and sensitizers. Semin Radiat Oncol 6:10-21.
Robinson SP, Rodrigues LM, Howe FA, Stubbs M, Griffiths JR (2001) Effects of different levels of hypercapnic hyperoxia on tumour R(2)* and arterial blood gases. Magn Reson Imaging 19:161-166.
Rockwell S, Dobrucki IT, Kim EY, Marrison ST, Vu VT (2009) Hypoxia and radiation therapy: past history, ongoing research, and future promise. Current Mol Med 9:442-458.
Rudat V, Vanselow B, Wollensack P, Bettscheider C, Osman-Ahmet S, Eble MJ, Dietz A (2000) Repeatability and prognostic impact of the pretreatment pO(2) histography in patients with advanced head and neck cancer. Radiother Oncol 57:31-37.
Saunders MI, Hoskin PJ, Pigott K, Powell ME, Goodchild K, Dische S, Denekamp J, Stratford MR, Dennis MF, Rojas AM (1997) Accelerated radiotherapy, carbogen and nicotinamide (ARCON) in locally advanced head and neck cancer: a feasibility study. Radiother Oncol 45:159-166.
Schuuring J, Bussink J, Bernsen HJ, Peeters W, van der Kogel AJ (2006) Effect of carbogen breathing on the radiation response of a human glioblastoma xenograft: analysis of hypoxia and vascular parameters of regrowing tumors. Strahlenther Onkol 182:408-414.
Schuuring J, Rijpkema M, Bernsen H, Bernsen P, van der Maazen R, Kaanders J, van der Kogel A, Heerschap A (2002) Effect of breathing a hyperoxic hypercapnic gas mixture on the oxygenation of meningiomas; preliminary results. J Neurooncol 57:127-132.
Smirnov AI, Norby SW, Clarkson RB, Walczak T, Swartz HM (1993) Simultaneous multi-site EPR spectroscopy in vivo. Magn Reson Med 30:213-220.
Song CW, Lee I, Hasegawa T, Rhee JG, Levitt SH (1987) Increase in pO 2
and radiosensitivity of tumors by Fluosol-DA (20%) and carbogen. Cancer Res 47:442-446.
Swartz HM, Walczak T (1998) Developing in vivo EPR oximetry for clinical use. Adv Exp Med Biol 454:243-252.
Thews O, Kelleher DK, Vaupel P (2002) Dynamics of tumor oxygenation and red blood cell flux in response to inspiratory hyperoxia combined with different levels of inspiratory hypercapnia. Radiother Oncol 62:77-85.
Williams KJ, Parker CA, Stratford IJ (2005) Exogenous and endogenous markers of tumour oxygenation status: definitive markers of tumour hypoxia? Adv Exp Med Biol 566:285-294.
Yaromina A, Kroeber T, Meinzer A, Boeke S, Thames H, Baumann M, Zips D (2011) Exploratory study of the prognostic value of microenvironmental parameters during fractionated irradiation in human squamous cell carcinoma xenografts. Int J Radiat Oncol Biol Phys 80:1205-1213.
HGH was responsible for the most aspects of the study including the data collection, data analysis and manuscript writing. NK was responsible for assisting in the design of the in vivo animal experiments and data analysis and preparation of paper publication. GXD provided assistance with data analysis and presentation of the data and preparation of paper publication. SH provided assistance with the presentation of the histological results. HS was responsible for analyzing and overviews and interpretation of the results. All authors have approved the final version of this paper for publication.
Conflicts of interest
The authors declare no conflicts of interest.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
[Table 1], [Table 2], [Table 3]
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