Tumor volume in subcutaneous mouse xenografts measured by microCT is more accurate and reproducible than determined by 18 F-FDG-microPET or external caliper. Tumor volume was measured with digital caliper as well. No, Is the Subject Area "Breast cancer" applicable to this article? Additionally, this technique is plagued by the inconsistent selection of tumor axes at subsequent examinations. Mice were monitored daily by personnel experienced in recognizing signs of morbidity (illness, injury, or abnormal behavior) also in weekends and holidays. Under these conditions, the formula for the volume of a tumor becomes. No, Is the Subject Area "Mouse models" applicable to this article? Case1 group members received one 200 g bevacizumab dose on the 8th day, case2 group members received 1.11 g bevacizumab from the 8th day every day for 15 days. The R2 value for the scanner is 0.99, as opposed to that for the caliper being 0.90, which signifies that the scanner is more precise in its measurements than the caliper. Cancer Res. However, if the treatment is lengthy, the problem of side-effects also has to be considered. Mice in the control group received no treatment (5 mice); case1 group members (10 mice) received 10 mg per kg body weight bevacizumab in one dose; while case2 group members (10 mice) received 1/180 dose of control dose for 15 days. 55, 11761180 (1995). The height and topography of the tumor is not taken into account in a caliper measurement. We have investigated the relationship between MRI-measured tumor volume and vascularization area (Phase III/3 case and control groups, 23rd (final) day of the experiment) but no significant correlation was found (same results were published in the case of Phase I [26] and Phase III/2 [23]). After sacrificing mice, tumors were removed, and their mass was measured. The authors declare no competing financial interests. The widely accepted method for calculating the volume of subcutaneous tumors is to measure the length and width of the tumor with a caliper, and, while assuming that the tumor is the shape of an ellipsoid and that the height of the tumor is equal to its width, use the formula for the volume of an ellipsoid. Usually, 3.0 x 10 6 cells are needed per injection. The TumorImagerTM line of scanners by Biopticon Corporation fixes these problems by keeping the animals relaxed during measuring and yielding measurements closer to actual values than caliper-based measurements. Hoffman-Goetz, L. et al. Bizberg Themes, By Memorial Sloan Kettering Cancer Center. J. Clin. Small MRI measurements were done in the Preclinical Imaging Center of Gedeon Richter Plc. Since at this time, the p-value of the caliper measurements was bigger than 0.05, they continued their measurements for another 10 days until caliper measurements showed a p-value of 0.0484 as shown in Figure 17. One can see from Table 2 that the goodness of the fit is different in the case of Phase I (tumor growth without therapy) and in the case of Phase III/3 (tumor growth with antiangiogenic therapy). https://doi.org/10.1038/laban.254. A caliper only digitizes the length and width of the tumor. Cancer Res. Traditionally, caliper measurements are used to calculate tumor volume. This process not only deforms the tumor mass but also creates undesirable pain on the animal. A smaller diameter mask will help to position the tumor in the center of the scan, but it will not alter this effect. 55, 74108 (2005). Harris, R.E., Alshafie, G.A., Abou-Issa, H. & Seibert, K. Chemopreventive of breast cancer in rats by celecoxib, a ciclooxygenase 2 inhibitor. Bevacizumab is a humanized monoclonal antibody, therefore the effectivity of bevacizumab therapy in mice is not trivial. Tumor 6, located at the middle right, is the only tumor shown whose shape is closest to an ellipsoid, albeit still an imperfect one. 8600 Rockville Pike This problem can be explained by our observation, namely tumors which were grown without therapy have more symmetric and solid closed shape, in contrast to tumors which were grown under antiangiogenic therapy. calipers and Water Displacement methods Tumor mimicking objects Vol. In the first subphase (Phase III/1), control group members (5 immunocompetent mice) received 10 mg per kg body weight bevacizumab, while case group members (5 immunocompetent mice) received one-tenth dose of control dose spread over 18 days. For small tumors, the error is about 7%, while for large tumors it is 27%.[3]. This work was supported by the Portuguese Foundation for Science and Technology (PTDC/DES/114122/2009 and Pest-OE/AGR/UI0772/2011 unity) and by the European Regional Development Fund (COMPETE, FCOMP-01-0124-FEDER-014707/022692). Results from a vaccine trial from AstraZeneca. Because tumor volume measurements used by researchers and industries are sensitive and often rely on the accuracy, the cutting-edge TumorImagerTM line of scanners is the ideal, most reliable choice for taking such sensitive measurements. In Phase IV, control group members received no bevacizumab. Comparison of volume measurement results between four users. The variables for the volume of a tumor, as measured by a caliper, are determined by a, b, and c in Figure 1, where length L is 2a, width W is 2b, and height H is c. VT, the volume of a tumor is determined by. Sci. If one measures the volume of a penny using calipers and uses the widely accepted formula, repeating the measurement each time another penny is added to the stack, the stack will yield the same volume each time, regardless of how many pennies are stacked. Figure 16. The aim of this article is therefore to explain of the tumor volume formula and the problems associated with the caliper measurements. Tumor was located in the last third of the back of the mouse in every case. Bethesda, MD 20894, Web Policies Similarly, Zhang et al. where v is tumor volume [mm3] and m is tumor mass [g]. Watch our 2-minute video overview. Open Access This approach presents an alternative to calipers which, although cheap, fast, and easy to use, introduce many biases for tumor volume estimation. Preclinical evaluation of LU 79553: a novel bis-naphthalimide with potent antitumor activity. We have administered 10 mg/kg body weight intraperitoneally, which means 200 g bevacizumab per a mouse, since the mass of the mice in the experiment was approximately 20 g (in the case of C57Bl/6 and SCID mice as well). Careers. Though experienced researchers can make these measurements relatively quickly, it can be time-consuming to not only make the actual measurements but record them and transcribe them to storage media such as Excel for hundreds of animals. Mice were humanely sacrificed using cervical dislocation. Therefore, optical imaging is . x|jY!==g'{oZ6-JiZ Then we calculated the Coefficient of Variance between such measurements for both measurements to compare average inter-operator differences. broad scope, and wide readership a perfect fit for your research every time. 2001 Jan 15;19(2):551-7. doi: 10.1200/JCO.2001.19.2.551. In addition, calipers also place undue strain on the animal during measurement, assume conditions for a perfect ellipsoid (which most tumors are not), and can lead to unnecessary losses of money over time. Copyright 2023 r>12TzPn 4x PUj50"pJ{{>y{.mn|Cvz{PP5x kH:5:YM.gn?c[J8oQWkR_!X/C@6P@WT=gO5Sn>om}&SI cyT>E"TnkhSju>~yVOm^C~0__ww@ y[xQr33?6O~hZcR{{u]=O5T. PLOS ONE promises fair, rigorous peer review, et al. In Phase III/3, control group contained 5 mice, case group contained 9 mice; all mice were implanted with C38 mouse colon adenocarcinoma. In the meantime, to ensure continued support, we are displaying the site without styles Quantity of the optimal solvent administration was also examined in this subphase. Notice that the caliper-based measurements have a larger standard deviation. 1 0 obj Figure 7. 105, 2023 (1983). Normality was investigated with one-sample Kolmogorov-Smirnov test, and homogeneity of variance (homoscedasticity) was examined with Levenes test. https://doi.org/10.1371/journal.pone.0142190, Editor: Francesco Bertolini, Treat. Epub 2014 May 28. Sport. Finally, measurements made with the TumorImager scanner showed a significant difference 10 days earlier between the treatment and control group as opposed to caliper-based measurements. The size of maximum tumor volumes observed in our study fulfills the requirements of Hungarian animal experimental research protocol. Correspondence to Clinical aspects which have to be considered to determine when animals should be euthanized are body weight, physical appearance, measurable clinical signs, unprovoked behavior and response to external stimuli. Mice in Phase III/3 control and case groups, and mice in Phase IV case1 and case2 groups received bevacizumab. Usage of this formula to calculate tumor volume from length and width values resulted in much more precise approximation of the MRI-measured tumor volume than protocol-based calculation. Pairwise comparison was done by Tukeys honest significant difference test to find those samples, which have significantly different means. In clinical practice, the determination of the tumor size (volume) is done by MRI and/or CT, but this is for the purpose of validating the effectiveness of the treatment, and therefore it is not a continuous monitoring. Clipboard, Search History, and several other advanced features are temporarily unavailable. 54, 401414 (1975). Stretching of the skin around the tumor of the mouse will cause an under-estimate of the tumor volume since the tumor is pressed into the body (see Figure 13. below). Kim HB, Zeng CH, Kim Y, Jeong S, Kim SH, Kang JM, Park Y, Won DS, Kim JW, Ryu DS, Lim B, Park JH. Mammary tumors seemed to take on an oblate spheroid geometry. The mathematical relationship between MRI-measured tumor volume and mass was investigated to estimate accurate tumor volume using caliper-measured data. A diagram of an ellipsoid, with the derivations of the variables needed to calculate its volume. The tumor number, from one to ten, is shown against the percent error. 16 (2008). Caliper Measurement. For more information about PLOS Subject Areas, click Contrast agents improve the visibility, but it is an extra strain to the organism, which can be lethal to animals which are in the final stage of cancer. Internet Explorer). In another, 7,286 tumors were measured and 99.97% of those exhibited smaller heights than widths. The figure shows the results of a mouse (C4) from control group (first row), and a mouse (E9) from case group (second row). Using a caliper, growth of the tumor in height is not measured as only width and length changes will affect the calculated volume. 2013 Oct;67(8):771-6. doi: 10.1016/j.biopha.2013.06.011. PhD from the 1st Department of Pathology and Experimental Cancer Research Institute from Semmelweis University, Budapest, for making the fluorescence pictures with confocal microscope. One common measure of efficacy is the T/C ratio, the ratio of tumor volume in control versus treated mice at a specified Substituting tumor mass values which were measured in Phase I into Eq 8, the corresponding tumor volume values can be evaluated (one can find numerical results in Table 2 Tumor volume MRI column; and graphical results in Fig 6). In that caseeven though all the three diameters can be measuredthe estimation of the volume has quite a large error. Ultrasonography as the Gold Standard for In Vivo Volumetric Determination of Chemically-induced Mammary Tumors. 2 0 obj Typical side-effects and adverse events are arterial and venous thromboembolic events, bleeding, hypertension, febrile neutropenia, infections, proteinuria, mucositis, and hand-foot syndrome. Hence, the first step has to be the vasculature normalization with the reconstruction of the balance of pro- and antiangiogenic factors. tumor height has to be approximated. Federal government websites often end in .gov or .mil. Due to the collaboration between medical doctors and biomedical engineers, model-based treatment protocols can be created. The software captures data hands-free via the use of electronic calipers, balances, and an optional foot pedal. Examining the relationship between MRI-measured tumor volume and tumor mass values, we have found a very strong linear correlation (R = 0.998, R2 = 0.996, p < 0.0001). Tumor volume measurement was performed by digital caliper and small animal MRI. Make sure you get accurate measurements for length, width and height. et al. Pritt, B., Ashikaga, T., Oppenheimer, R.G. Thank you for visiting nature.com. %PDF-1.4 One of them died on the 18th day, and another one on the 23rd day. Substituting tumor mass values which were measured on the 24th day of Phase I to the equation of the resulted curve, the corresponding tumor volume values can be evaluated (evaluated data). In addition, the mice were weighed . Provided by the Springer Nature SharedIt content-sharing initiative, Bulletin of the National Research Centre (2022), Lab Animal (Lab Anim) In this study, the authors measured dimensions of rat mammary tumors using a caliper and using real-time compound B-mode ultrasonography. J. Natl. No, PLOS is a nonprofit 501(c)(3) corporation, #C2354500, based in San Francisco, California, US, Corrections, Expressions of Concern, and Retractions, https://doi.org/10.1371/journal.pone.0142190, http://www.gene.com/download/pdf/avastin_prescribing.pdf, http://www.ema.europa.eu/docs/en_GB/document_library/EPAR_-_Scientific_Discussion/human/000582/WC500029262.pdf, http://www.protocol-online.org/prot/Protocols/Xenograft-Tumor-Model-Protocol-3810.html, http://www.targeson.com/sites/default/files/content/pages/pdfs/tail_vein_protocol_2012_0.pdf. The manipulation of the handheld tool is at least as easy as holding a measurement caliper. Even though, now, the formula is calculated in an Excel spreadsheet where can be defined as a function, pi(), almost in all cases, in the volume formula is defined as 3.0. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). 17, 905910 (2004). As is the case with caliper measurements, if the tumor appears visibly flattened and is hard to see by eye then the mouse should be repositioned. B){00)Y$Ixlr{dYe}#@ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ ,^X_zN5:EZN]zM^k /Wu^ZZi#Ju.!L!z@u*kUim|+aZmjth-}z5'^|= 4L1_]qVZV*h@! BMC Medical . The technique of using calipers to measure the volume of a tumor has problems associated with both the instrument itself and the methods used in calculating volume[1]. In order to estimate tumor volume by external caliper, the greatest longitudinal diameter (length) and the greatest transverse diameter (width) were determined while mice were scruffed and conscious. In the light of the above mentioned, linear curve fitting was carried out to find the mathematical relationship between MRI-measured tumor volume and tumor mass (Phase III/3 case and control groups). 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