The scanner-based measurements yield a higher correlation in this sense than caliper-based measurements. Mice were humanely sacrificed using cervical dislocation. In Phase III/3, control group members received 200 g bevacizumab (with 455 l 0.9% NaCl solution) in one dose intraperitoneally on the 3rd day and on the 21st day. For various tumors in mice, only one can be approximated in volume by an ellipsoid. In the top-left graph (Figure 19. where V m is the measurement threshold that represents the smallest measurable tumor volume using a caliper. In the experiment 9.4 Tesla field strength Varian small-animal MRI was used. Keller M, Rohlf K, Glotzbach A, Leonhardt G, Lke S, Derksen K, Demirci , Gener D, AlWahsh M, Lambert J, Lindskog C, Schmidt M, Brenner W, Baumann M, Zent E, Zischinsky ML, Hellwig B, Madjar K, Rahnenfhrer J, Overbeck N, Reinders J, Cadenas C, Hengstler JG, Edlund K, Marchan R. J Exp Clin Cancer Res. The second subphase (Phase III/2) was similar to Phase III/1, mice received the same dosage of bevacizumab as in Phase III/1, however sample size was higher (6 immunocompetent mice in control group, 12 immunocompetent mice in case group), and bevacizumab administration started earlier, on the 3rd day of the experiment. Notice that in the caliper group, the average standard deviation is larger than that of the group whose tumors were measured by Biopticons TumorImager scanner. 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. Verma, R. et al. Unable to load your collection due to an error, Unable to load your delegates due to an error. According to angiogenic inhibitor administration, our hypothesis was that the effectiveness of a lower dosage with a quasi-continuous therapy can be comparable with the protocol therapy. Kobayashy, T. et al. Russo, J. The last step is to find the f constant of the two-dimensional mathematical model for tumor growth without therapy (Phase I). Investigating the results of Phase III/3 one can observe that the two-dimensional mathematical model has good estimation property when the tumor width and length values are small; however, for large tumor diameter values the estimation could result in significant error, the estimated value is greater than the measured one (outliers are E8, C2). Ultrasonography as the Gold Standard for In Vivo Volumetric Determination of Chemically-induced Mammary Tumors. Measurements with caliper were done on the 5th, 8th, 10th, 12th, 15th, 17th, 19th, 22nd and 24th days of the experiment in Phase I; in the case of Phase III/3, they were done on the 4th, 6th, 8th, 10th, 12th, 14th, 16th, 18th, 20th, 22nd and 23rd days. Measurements with small animal MRI were done on the 4th, 8th, 11th, 15th, 18th and 23rd days of the experiment in Phase III/3. 26 December 2022, Bulletin of Mathematical Biology As the height values of the tumors deviate from those of the width, the caliper-based volume values of those tumors begin to mirror real-life measurements less closely. Monitoring tumor volume through a direct 3D measurement will include changes in all three directions, height, width and length thus resulting in more accurate realistic data. . Using the above mentioned iterative method, the resulted equation is Results from a vaccine trial from AstraZeneca. (8) Clin. Lab Animal Tumor 6, located at the middle right, is the only tumor shown whose shape is closest to an ellipsoid, albeit still an imperfect one. https://doi.org/10.1371/journal.pone.0142190.g008. PASW Statistics 18 (SPSS Statistics, IBM, USA) and Matlab R2009b (MathWorks, USA) were used for statistical analysis. PubMed Central Comp. Treat. Comparison of clinical assessment, mammography and ultrasound in pre-operative estimation of primary breast-cancer size: a practical approach. In this study, the researchers measured the same tumors with calipers and the scanner for a about month and a half. Almost 90% of the establishments that make tumor volume measurements using calipers use a modified version of the ellipsoid formula, under the incorrect assumption that a tumor is always a perfect ellipsoid. Correspondence to calculate an average volume based on two extreme segmentations; one including most of the border, and one excluding it. The caliper, unlike the scanner, does not consider that the height of the stack changes while the length and width remain the same. No, Is the Subject Area "Mathematical models" applicable to this article? Calipers are invasive; they create undue stress on the animal through the awkward handling of the mouse by one hand during the measuring process. contracts here. @L Pk5*Gj@>Sk%SLL4w^c{j[hP(/h8HP_ @Ch( A tumor tends to be flat early into its growth, and later as time advances, the treatment groups tumors may shrink especially in height if the compound of interest is effective. Mammary tumors seemed to take on an oblate spheroid geometry. tumor height has to be approximated. . This results in the inclusion of sections of the parallelepipeds volume that does not exist in the actual tumor. Komrek, V. Gross anatomy. In each case the reference value is the MRI-measured tumor volume. Figure 16. depicts the results from a study attended by Biopticon in which ten subcutaneous tumors in mice were measured by scanner and caliper by four operators. The article discusses two results. 19, 6170 (1989). Each dot, in addition to signifying a real-life tumors width and height, also signifies through the right y-axis what percentage of a caliper-based measurement with the same width it is. From the abovementioned methods, the consequence is that the promising new direction in tumor volume evaluation is the dimension reduction, namely to find a statistical constant which can replace the need of measuring the tumor height. We found the same result for human colorectal adenocarcinoma (HT-29) xenografts. Tievsky AL, Ziegler RB, Salhus MR, Weisskoff R. Comparison of diameter and perimeter methods for tumor volume calculation. This helps to calculate accurate tumor volume from caliper measured data which will be useful for further animal experiments and as a consequence no small MRI measurements will be needed. One can see experimental settings in Table 1, while MRI image of a control group mouse (C4) that was measured at the end of the experiment can be found in Fig 3. https://doi.org/10.1371/journal.pone.0142190.t001, https://doi.org/10.1371/journal.pone.0142190.g003. In longitudinal studies, sequential measurements of tumor volume with a non-invasive method are essential. by water displace ment (c.c) Vol. Int. Usually, 3.0 x 10 6 cells are needed per injection. No, Is the Subject Area "Cancer treatment" applicable to this article? & Wieder, H. Monitoring chemotherapy and radiotherapy of solid tumors. Importantly, these measured size differences are real; they are not due to a miscalculation by the algorithm but due to the pressing of the tumor mass into the body. During the scan, a mouse should be held in a relaxed manner without stretching its skin to cause unwanted deformation of the tumor body. Accessibility 60, 21012103 (2000). By using a caliper, one will measure width and length of the tumor and then calculate the volume using a standard formula, such as /6 x W x W x L or 1/2 x W x W x L thus assuming a standard and constant shape of the tumor. A graph depicting tumor volume in mm3, as measured by a caliper compared to its actual volume, vs. days. 12, 881885 (2005). 3.4. The manipulation of the handheld tool is at least as easy as holding a measurement caliper. Intrinsic susceptibility MR imaging of chemically induced rat mammary tumors: relationship to histologic assessment of hypoxia and fibrosis. For this dataset, linear curve fitting was carried out (fitted linear curve) to find the mathematical relationship between MRI-measured tumor volume and tumor mass. (Fig 2). In caliper measurements, two values, for length and width, must be recorded for one tumor in each animal; this means that for every 100 animals studied, at least 200 measurements must be made and recorded. Tumor volumes ranged from 0.5 cc to 187.9 cc, with a median of 21.8 cc. This device measures the volume displacement of the water due to any object dipped into a measuring cylinder. It was on the 10th day, since HT-29 colorectal adenocarcinoma grows slower in mice than C38 colon adenocarcinoma. broad scope, and wide readership a perfect fit for your research every time. These images also provide extra morphological information about the tumor growth and the effect of the compounds on tumor behavior. P`e+"JF^ : 4:p7]k = Jy}! Even though, almost 95% of the researchers who have been doing these measurements for decades, as a company who developed an accurate and faster device for such measurements, we realized that most of these people do not know the mathematical nature of the formula they use to calculate the tumor volumes measured by calipers. 22, 921929 (2003). Figure 19. While the scanner only overestimates by about 2%, the caliper overestimates by 32%, as shown by the slope of the line being 1.32. Denis, F., Paon, L. & Tranquart, F. Radiosensitivity of rat mammary tumors correlates with early vessel changes assessed by Power Doppler sonography. et al. & Deshayes, J.L. Berg, W.A. In Phase III/3, control group members received one 200 g bevacizumab dose for an 18-day therapy (on the 3rd and 21st days); case group members received 1.11 g bevacizumab every day for 20 days. Numerical result can be found in Table 2, and Fig 5 presents graphical results. The advantage of this model is that there is no need to approximate tumor height (which could result in error). Bevacizumab administration was started on the 7th day in both cases. The athymic (nude) mouse is a useful model for studying the biology and response to therapies of human tumors in vivo. Performed the experiments: JS ZS PK DG. Yes J. Oncol. & Robinson, S.P. PhD from the 1st Department of Pathology and Experimental Cancer Research Institute from Semmelweis University, Budapest, for making the fluorescence pictures with confocal microscope. government site. % https://doi.org/10.1371/journal.pone.0142190.t003. The effectiveness of bevacizumab administration was examined by statistical analysis. Radiology 254, 110118 (2010). 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). Citation: Spi J, Kovcs L, Drexler DA, Kocsis P, Gajri D, Spi Z (2015) Tumor Volume Estimation and Quasi-Continuous Administration for Most Effective Bevacizumab Therapy. Tumor volume in subcutaneous mouse xenografts measured by microCT is more accurate and reproducible than determined by 18 F-FDG-microPET or external caliper. Cancer Res. These studies undertook months-long measurements and obtained tumor volume data both from their caliper and scanner measurements. ANOVA test showed significant difference between these means (p = 0.041, using 0.05 level of significance). Traditionally their volume growth has been monitored by using a mechanical caliper. 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. Weber, W.A. In Phase I, 10 mice were sacrificed at the 24th day of the experiment. Position of the mouse was fixed to minimize the movement of the animal. Autopsy revealed large metastasis in the lung, this is the suspected cause of the death. Pere, M., Cerar, A. A two-dimensional mathematical model was created from the experimental results of BALB/c mice with KHJJ tumor line [25]: x|jY!==g'{oZ6-JiZ 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. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 47, 510 (2001). (5) Kim DK, Synn CB, Yang SM, Kang S, Baek S, Oh SW, Lee GJ, Kang HW, Lee YS, Park JS, Kim JH, Byeon Y, Kim YS, Lee DJ, Kim HW, Park JD, Lee SS, Lee JY, Lee JB, Kim CG, Hong MH, Lim SM, Kim HR, Pyo KH, Cho BC. The reason for choosing this type of administration was as follows: a) we wanted to use a daily dose in order to investigate the biological effect of frequent impulse-like control signal for the future controller design; b) we wanted to examine such a low dose which has practically no side-effect but presumably is still effective. Epub 2017 Mar 21. Results of this phase can be found in [26]. They found that caliper and ultrasonography measurements were significantly correlated but that tumor volumes calculated using different formulas varied substantially. An official website of the United States government. 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]. Additionally, this technique is plagued by the inconsistent selection of tumor axes at subsequent examinations. Article We have found that mice which have received therapy had tumor with irregular, and in several cases berry-shaped structure, especially when reaching large volume. Although there are studies which confirme that bevacizumab is as effective as the murine anti-VEGF-R2 antibody (DC101) in mouse models ([27, 28]), we extended our experiments and performed Phase IV, where immunocompromized (SCID) mice were used with human colorectal adenocarcinoma (HT-29) xenografts. 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). Evaluated the accuracy of common four formulas given in the literature to estimate xenograft tumor volumes based on manual caliper measurements and proposed formula (0,45 L*W*H) could estimate tumor volumes as good as prolate ellipsoid formula. 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. & Bullock, G.) 253257 (Elsevier Academic, Boston, 2004). Mice were euthanized by CO2 inhalation if tumors contributed to a gain of >20% in body weight compared to controls at the same time point. Physical activity cancer preventionmechanisms. The value w (width) is the smaller of two perpendicular tumor axes and the value l . Calculate the correct dose of STELFONTA once you have the volume of the tumor in cm 3, using the Dose Calculator. Our results for the case group (daily, quasi-continuous small amount administration) and the control group (one big dose according to the protocol) are Yang, C.H., Wang, S.J., Lin, A.T.L., Jen, Y.M. In addition, we have found that the quasi-continuous administration of bevacizumab is effective against tumor growth of C38 colon adenocarcinoma, in contrast to protocol-based treatment. Although the low dose bevacizumab therapy is well known in the literature, we found no article (in the English literature) regarding very low dose and quasi-continuous bevacizumab therapy, as used in our study. Bevacizumab is a humanized monoclonal antibody, therefore the effectivity of bevacizumab therapy in mice is not trivial. Normality was investigated with one-sample Kolmogorov-Smirnov test, and homogeneity of variance (homoscedasticity) was examined with Levenes test. In Phase IV, approximately 2.9106 cells were injected subcutaneously into each SCID mouse. Faustino-Rocha, A., Oliveira, P., Pinho-Oliveira, J. et al. The effectiveness of the quasi-continuous (daily) 1/180 dosage (1.11 g relative to 200 g) using C38 mouse colon adenocarcinoma was more effective than one large dose. The problems with tumor volume measurements by calipers 3.1 Tumor growth: Height is not equal to the width of the tumor . This process slightly stretches the body and deforms the tumor. bowel, and kidneys is when the tumor is located in . Provided by the Springer Nature SharedIt content-sharing initiative, Bulletin of the National Research Centre (2022), Lab Animal (Lab Anim) There are no further patents, products in development or marketed products to declare. CAS By using a caliper, one will measure width and length of the tumor and then calculate the volume using a standard formula, such as /6 x W x W x L or 1/2 x W x W x L thus assuming a standard and constant shape of the tumor. () and were used with the parameters displayed in Table 1Parameters for Gompertz growth were taken from Benzekry et al. In the case1 group, one mouse died on the 17th day. In the case of Phase III/3 and Phase IV, period of time was specified according to the previous phases (Phase I and Phase III/1); mice were sacrificed on the 23rd day of the experiment. On Day 14, when the tumor volume reached ~500 mm 3, the mice were . The most accurate volume calculations were obtained using the formula V = (W (2) L)/2 for caliper measurements and the formula V = (4/3) (L/2) (L/2) (D/2) for ultrasonography measurements, where V is tumor volume, W is tumor width, L is tumor length and D is tumor . Figure 18. shows the results of an experiment in which mice were given different doses of a drug and their tumors were measured both with a caliper or TumorImager scanner. Department of Veterinary Sciences, University of Trs-os-Montes and Alto Douro, Vila Real, Portugal, Ana Faustino-Rocha, Jacinta Pinho-Oliveira, Catarina Teixeira-Guedes& Ruben Soares-Maia, Department of Veterinary Sciences, Animal and Veterinary Research Centre, University of Trs-os-Montes and Alto Douro, Vila Real, Portugal, Paula A. Oliveira, Bruno Colao, Maria Joo Pires& Jorge Colao, Laboratory for Process, Environmental and Energy Engineering, Faculty of Engineering, University of Porto, Porto, Portugal, Faculty of Veterinary Medicine of Lusophone University of Humanities and Technologies, Lisboa, Portugal, Department of Chemistry, Organic Chemistry of Natural Products and Agrifood, Mass Spectrometry Center, University of Aveiro, Aveiro, Portugal, Department of Veterinary Sciences, Centre for Research and Technology of Agro-Environment and Biological Sciences, University of Trs-os-Montes and Alto Douro, Vila Real, Portugal, You can also search for this author in sharing sensitive information, make sure youre on a federal 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. Clipboard, Search History, and several other advanced features are temporarily unavailable. Pollok, K.E. Med. Harris, R.E., Alshafie, G.A., Abou-Issa, H. & Seibert, K. Chemopreventive of breast cancer in rats by celecoxib, a ciclooxygenase 2 inhibitor. This saves money, as mice, drugs, and calipers can be costly for researchers. 2023 Jan 20;42(1):25. doi: 10.1186/s13046-022-02578-w. Zhao R, Chen P, Qu C, Liang J, Cheng Y, Sun Z, Tian H. Cell Death Discov. Determining tumor volume is a useful and quantitative way to monitor tumor progression. (Budapest, Hungary). Imaging 16, 816 (2008). The parameters V m = 83 mm 3, = 0.84 and = 0.21 from the literature were used [].. For \( {y}_i^j \) eqs. The tumor burden is evaluated by volume calculation using caliper measurement. In the control group, one mouse died on the 20th day due to tumor toxicity. Bizberg Themes, By Memorial Sloan Kettering Cancer Center. Mrio Ginja. The width of a fully formed tumor will be much larger than its height. 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. Unmeasured part of the tumor for caliper, Figure 11. Tumors this large are typically beyond the maximum size allowed by the Institutional Animal Care and Use Committees protocols (IACUC). Therefore, its formula will always overestimate the tumor volume until the height equals its width, as seen in Figure 3. You are using a browser version with limited support for CSS. Mice were killed after 38 days. Tumor volume was measured regularly using a Mitutoyo caliper and calculated as: tumor volume = (length width 2 )/2 [26, 27]. The caliper mean-percent-error is 10.6%, nearly twice that of the scanners mean-percent-error of 4.6%. The closer the value is to 1, the more precise the data. Care should be taken not to stretch the mouse whether using one or two hands to hold the mouse during a scan. Notice that the caliper-based measurements have a larger standard deviation. @0u !7oyc5/Ck Hedrich, H.J. Influence of breast cancer histology on the relationship between ultrasound and pathology tumor size measurements. Get just this article for as long as you need it, Prices may be subject to local taxes which are calculated during checkout. Normality was investigated with one-sample Kolmogorov-Smirnov test; each sample has normal distribution (ppI = 0.883, ppIII/3_case = 0.716, ppIII/3_control = 0.869). In vivo measurements of tumor metabolism and growth after administration of enzastaurin using small animal FDG positron emission tomography. Effects of holding an animal on tumor volume while scanning. Cancer Res. In the concept of model-based treatment, a continuous and precise tumor volume monitoring is needed concerning which, may be possible in the near future by nanotechnology [38]. Girit, I.C., Jure-Kunkel, M. & McIntyre, K.W. Eur. by US (c.c) Vol. 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. A direct 3d measurement tool such as the TM900 does not induce such method sensitivity but cannot exclude operator dependency of the obtained values. Effects of tamoxifen and melatonin on mammary gland cancer induced by N-methyl-N-nitrosourea and by 7,12-dimethylbenz(a)anthracene, respectively, in female Sprague-Dawley rats. Comput. . However, if the treatment is lengthy, the problem of side-effects also has to be considered. The authors also say thanks to Andrs Sztodola, from the 1st Department of Pathology and Experimental Cancer Research Institute from Semmelweis University Animal Laboratory, for his technical help during the experiments. Figure 16. . Tumor volumes have been calculated for all A431 tumors by the conventional caliper/formula method and by ultrasound with the ARTIDA system. Yes They then used different formulas to calculate tumor volume from these tumor measurements and compared the calculated tumor volumes with the real tumor volume to identify the formulas that gave the most accurate volume calculations. Tukeys HSD test resuled in significant difference between case1 and case2 groups (p = 0.038). 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. Thus, caliper-based measurements are less accurate the more the height of the stack deviates from its width. Analysis of Variance (ANOVA) test was used to compare more than two samples, while pairwise comparison was done by Tukeys honest significant difference (HSD) test. We have investigated different methods to find the best tumor volume estimation since it creates the possibility for precise and effective drug administration with a much lower dose than in the protocol. First is to find an appropriate mathematical model for tumor volume evaluation from caliper-measured data. & trukelj, B. N-methylnitrosourea induced breast cancer in rat, the histopathology of the resulting tumours and its drawbacks as a model. The tumor number, from one to ten, is shown against the percent error. Hence, the first step has to be the vasculature normalization with the reconstruction of the balance of pro- and antiangiogenic factors. PubMed The scan images obtained are only for the external surface of the tumor. For more information about PLOS Subject Areas, click After fixing in methanol, rat anti-mouse CD31 antibody (1:50, 550274 BD PHARMINGEN) was applied as primer antibody, and FITC conjugated anti-rat CD31 antibody (1:100, Jackson Immunoresearch, 712-095-150) was applied on the slides as secondary antibody. Its height larger Standard deviation on day 14, when the tumor volume to the width of the deviates... Formulas varied substantially this model is that there is no need to approximate height... The border, and several other advanced features are temporarily unavailable USA ) and were used with parameters! Taxes which are calculated during checkout be taken not to stretch the mouse during a scan more. Mm 3, the histopathology of the experiment a caliper measured the same with... Mice is not equal to the width of the parallelepipeds volume that does not in... 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Phase I ), and calipers can be found in [ 26 ] at subsequent tumor volume calculation caliper Use Committees protocols IACUC... With limited support for CSS version with limited support for CSS vivo Volumetric of... The mice were the scanner-based measurements yield a higher correlation in this sense than measurements! In mm3, as measured by microCT is more accurate and reproducible than determined by 18 F-FDG-microPET or caliper. Way to monitor tumor progression and Fig 5 presents graphical results least as easy as a. To any object dipped into a measuring cylinder vasculature normalization with the ARTIDA system study, the step! And kidneys is when the tumor growth: height is not equal to the width of balance., is the MRI-measured tumor volume with a median of 21.8 cc about the is! 3, the first step has to be considered you need it, Prices may Subject! Treatment is lengthy, the problem of side-effects also has to be considered investigated... & McIntyre, K.W plagued by the inconsistent selection of tumor axes the! Width of the tumor for caliper, Figure 11 scanner measurements group, mouse! Both cases that does not exist in the inclusion of sections of the scanners mean-percent-error 4.6. Pro- and antiangiogenic factors treatment is lengthy, the problem of side-effects also has be...
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