Animals were carried out in the most humane and environmentally sensitive manner possible; in addition the 3Rs principle (replacement, refinement, reduction) was adequately implemented according to the Directive 2010/63/EU of the European Parliament. A two-dimensional mathematical model was applied for tumor volume evaluation, and tumor- and therapy-specific constants were calculated for the three different groups. Figure 4. Lab Anim 42, 217224 (2013). Kubatka, P. et al. To reduce the time of analysis one could e.g. Experimentally induced mammary tumors in rats. Mice were humanely sacrificed using cervical dislocation. The tumor volume calculation was executed using the formula V = (W2 L)/2 for caliper measurements, where V is tumor volume, W is tumor width, and L is tumor length . When using a handheld direct 3D measurement instrument such as the Peira TM900, the obtained value is independent from the angle position of the tool on the tumor. With constant width and length, the measured volume changes with the shape of the tumor. Influence of breast cancer histology on the relationship between ultrasound and pathology tumor size measurements. Kim HB, Zeng CH, Kim Y, Jeong S, Kim SH, Kang JM, Park Y, Won DS, Kim JW, Ryu DS, Lim B, Park JH. https://doi.org/10.1038/laban.254. It can be carried out in vivo during the experiment due to the subcutaneous localization of the tumor. Autopsy revealed only minimal local tumor; however death was not caused by tumor. No, Is the Subject Area "Magnetic resonance imaging" applicable to this article? Using a caliper, growth of the tumor in height is not measured as only width and length changes will affect the calculated volume. https://doi.org/10.1371/journal.pone.0142190.g007. That is independent from the use of a direct optical 3D measurement tool or a caliper. Google Scholar. 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. These model-based protocols can be more effective than the current ones, since they provide individual treatment for the patients. One can see that the two-dimensional mathematical model fits to the MRI-measured values, while the protocol-based values present totally different curve. P`e+"JF^ : 4:p7]k = Jy}! Hoffman-Goetz, L. et al. When the tumor has a symmetrical, solid closed shape (typically without treatment), volume can be evaluated accurately from caliper-measured data with the applied two-dimensional mathematical model. Ergonomics 37, 217229 (1994). The caliper mean-percent-error is 10.6%, nearly twice that of the scanners mean-percent-error of 4.6%. The cells are forced to grow laterally because of the surface tension of the skin above, and the flesh below, which therefore expand the width of the tumor length and widthwise rather than along its height. In one trial, 208 tumors were measured, and 98.94% of those tumors exhibited much smaller heights than widths. A solution to these problems is the TumorImagerTM scanner. In the case of Phase I, no similar problem occurs; the two-dimensional mathematical model can handle great values as well (e.g. For example, one of the main reasons why the US FDA revoked the approval of Avastin for treating advanced breast cancer is the high rate of side-effects when bevacizumab was applied [46]. During the MRI measurements, life parameters of the mice were monitored. contracts here. 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. J. Ultrasound Med. where v is tumor volume [mm3] and m is tumor mass [g]. & Lin, C.A. J. Urol. depicts the inaccuracy of the formula used for caliper measurements, in which the height of the tumor is assumed to be equal to its width. In Phase IV, eleven weeks old male SCID mice with implanted HT-29 human colorectal adenocarcinoma was used. . In order to assess the accuracy of tumor measurements by the scanner, we measured the volume of many tumors with a caliper (VC) and with the scanner (VS) first, then after terminating the animals we dissected tumors right around the edges and measured their volumes by dipping them in an accurate volume (VPleth) measuring device called Plethysmometer[8]. In order to assess interobserver variation, each . The article discusses two results. One can find datasets in the last column of Table 2. Monaco performed the dose calculation of the VMAT treatment plans with the dose calculation grid of 3.0 mm by utilizing the Monte Carlo algorithm. Cancer Res. 120, 386391 (2003). In the case of Phase I and Phase IV, there were no MRI measurements. Am. Conceived and designed the experiments: ZS JS. The effectiveness of bevacizumab administration was examined by statistical analysis. McCormick, D.L., Adamowski, C.B., Fiks, A. & Pisani, P. Global cancer statistics, 2002. Google Scholar. Tukeys HSD test resuled in significant difference between case1 and case2 groups (p = 0.038). Yes Performed the experiments: JS ZS PK DG. A patented algorithm running in the TumorManagerTM program calculates the tumor volume by creating a 3D surface map of the tumor (red-colored surface in Figure 12.). CAS Yes Biol. A typical comparison of tumor volume measurements by TumorImager-2TM with calipers. 12 immunocompetent mice were transplanted subcutaneously with C38 colon adenocarcinoma, and 11 immunocompetent mice were injected intramuscularly with B16 melanoma. Caliper Measurement. McPhail, L.D. Faustino-Rocha, A., Oliveira, P., Pinho-Oliveira, J. et al. 2 0 obj Verma, R. et al. 35, 18341840 (2003). Med. Copyright 2023 On the other hand, values given by a scanner (VS, red triangles) more closely follow the true volume of the pennies (VT, blue diamonds). Tumor Volume Measurement Treat. 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary. This saves money, as mice, drugs, and calipers can be costly for researchers. calipers and Water Displacement methods Tumor mimicking objects Vol. In the case2 group, one mouse died on the 8th day, before the therapy (bevacizumab administration) started. (3), Another relatively new but not widely used approach to estimate tumor volume is to assume hemi-ellipsoid shape [24]. 17, 905910 (2004). x|jY!==g'{oZ6-JiZ 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). Pathology of tumours in laboratory animals. PLoS ONE 10(11): 20, the red dots that represent scanner-based measurements are closer to the black dotted (objective) line than the blue squares that represent caliper-based measurements. 01 July 2022. Figure 19. 19, 6170 (1989). 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. We compared the volumes, areas . To evaluate tumor volume, first the shape of the tumor has to be assumed, and if it is necessary for volume calculation, the third diameter (height, h) has to be approximated. The aim of this article is therefore to explain of the tumor volume formula and the problems associated with the caliper measurements. Both methods are comparable when it concerns the time it takes to do a measurement. Controller-based therapy has the obvious advantage against e.g. In the experiment 9.4 Tesla field strength Varian small-animal MRI was used. Calipers are invasive; they create undue stress on the animal through the awkward handling of the mouse by one hand during the measuring process. In a 2001 study utilizing MRI, it was concluded that 85% of tumors needed a measurement other than length and the width to yield an accurate volume[5]; caliper-based values do not provide this other measurement. 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. Measure the mass by using a balance (0.01g) and compute the volume by using the et al. Imaging 16, 816 (2008). An average penny has a diameter of 19 mm and a height of 1.27 mm, yielding a volume of 360.1 mm3. In addition, the mice were weighed . Unauthorized use of these marks is strictly prohibited. The authors declare no competing financial interests. The problems with tumor volume measurements by calipers 3.1 Tumor growth: Height is not equal to the width of the tumor . & Russo, I.H. Department of Control Engineering and Information Technology, Budapest University of Technology and Economics, Budapest, Hungary, Affiliation Is the Subject Area "Cancers and neoplasms" applicable to this article? Preclinical evaluation of LU 79553: a novel bis-naphthalimide with potent antitumor activity. The true volume (VT) of a stack of pennies as measured by caliper (VC) and scanner (VS). The TM900 also stores an optical image in the measurement data base. Bookshelf This formula was the starting point of our examination to find an appropriate mathematical model. Before produced by the TumorImager-2TM so that the animal is aligned under the mask each time similarly to the previous scan. Examination of tumor growth belongs not only to the basic medical research, but to the biomedical engineering field as well. PLOS ONE promises fair, rigorous peer review, First of all, our hypothesis according to the importance of administration is proved. et al. Measured data is the MRI-measured tumor volumetumor mass pairs on the 23rd day of Phase III/3 (case and control group). Therefore, optical imaging is . Figure 14. & Wieder, H. Monitoring chemotherapy and radiotherapy of solid tumors. 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). Unmeasured part of the tumor for caliper, Figure 11. Thus, caliper-based measurements are less accurate the more the height of the stack deviates from its width. To compare more than two samples, ANOVA test was applied. Another approach is to utilize the scan images (Figure 14.) Read the article on why you should not use calipers! Google Scholar. Cancer Res. The problems with tumor volume measurements by calipers, 3.1 Tumor growth: Height is not equal to the width of the tumor, 3.3 Caliper measurements are time-consuming, 3.4 Caliper measurements ignore a larger volume, https://ugobasile.com/products/catalogue/pain-and-inflammation/plethysmometer, A Structured Light Based Laboratory Animal Subcutaneous Tumor Scanning System. In Phase III/3, case group members received one-tenth dose of control dose intraperitoneally spread over 18 days. 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. 2022 Dec 5;10:998013. doi: 10.3389/fchem.2022.998013. Figure 13. 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. Several studies have investigated the effectiveness of bevacizumab therapy according to different cancer types but these days there is an intense debate on its utility. The defined shape of an actual tumor (red) and the shape measured by a caliper (blue box). Tumor volume was calculated by the modified ellipsoidal formula: V = (Length Width 2) . Figure 17. J. Nucl. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). To obtain Even though we did not have a statistical analysis of volume loss due to the unmeasured part of tumors in caliper measurements, the lost volume could be higher than the scanner results. It means that knowing the tumor mass, tumor volume can be estimated with suitable accuracy; hence the lack of MRI measurement can be replaced in the case of Phase I. Care should be taken not to stretch the mouse whether using one or two hands to hold the mouse during a scan. [45] have found that the combination of low-dose cyclophosphamide and ginsenoside Rg3 therapy can be more effective than the normal administration. For more information about PLOS Subject Areas, click Where anesthesia was used (2%-4% isourane in room air, 1 L/min), body temperature and respiration rate were maintained at 37 C and 60-90 breaths/minute, respectively. For the most precise result, tumor mass values, which were measured at the end of the experiment, were compared in the statistical analysis. 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. Figure 6. 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). The fractional change of the measured tumor volume is greater in the case of small tumors when their height is only a few millimeters. by US (c.c) Vol. The 1st Department of Pathology and Experimental Cancer Research of the Semmelweis University (Budapest, Hungary) and the Physiological Controls Group of the Obuda University (Budapest, Hungary) began collaborating on antiangiogenic therapy research in 2012including the current experimental investigations. Firstly, since the tumor is assumed to be an ellipsoid shape, and the tumors morphology often deviates significantly from this as it grows, this method will produce errors as big as 18.4%. Bethesda, MD 20894, Web Policies This device measures the volume displacement of the water due to any object dipped into a measuring cylinder. Finding the mathematical relationship between MRI-measured tumor volume and tumor mass creates the possibility to estimate tumor volume from caliper-measured data. <> No, Is the Subject Area "Renal cancer" applicable to this article? . This paper reviews the standard technique for tumour volume assessment, calliper measurements, by conducting a statistical review of a large dataset consisting of 2,500 tumour volume measurements from 1,600 mice by multiple operators across 6 mouse strains and 20 tumour models. In Phase I, 10 mice were sacrificed at the 24th day of the experiment. The specific roles of these authors are articulated in the author contributions section. https://doi.org/10.1371/journal.pone.0142190.g002. Tumor volumes ranged from 0.5 cc to 187.9 cc, with a median of 21.8 cc. The site is secure. Modern Pathol. The whole experiment consisted of four phases; the article focuses on the Phase III/3 and its results together with the comparison of Phase I and Phase III/3 (Fig 1). 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. Gullino, P.M., Pettigrew, H.M. & Grantham, F.H. Girit, I.C., Jure-Kunkel, M. & McIntyre, K.W. Determining tumor volume is a useful and quantitative way to monitor tumor progression. These measurements gave us accurate tumor volumes to compare the accuracy of this new technique in measuring tumor volumes. Accuracy of Optical Imaging for Measuring Tumor Burden In Vivo . In addition, the gap between tumor volume values for caliper-based control and treatment groups is smaller than that for scanner measurements; the significant p-value was over 50 times higher when using calipers (p > 0.05) than with scanners (p < 0.001), suggesting that the caliper-based measurements are not statistically significant. A standard deviation of 0% is almost impossible in real life, but a smaller standard deviation is generally favorable. Studies using other methods of imaging have exposed the problems that calipers pose. An official website of the United States government. & Hafstrm, L. Estimation of liver tumor volume using different formulasan experimental study in rats. by water displace ment (c.c) Vol. The effectiveness strongly depends on the administration, and a drug which is effective on a molecular level can be applied in a less effective way because of the incorrectly chosen administration. The last step is to find the f constant of the two-dimensional mathematical model for tumor growth without therapy (Phase I). BMC Medical . In order to reduce such deformations on the tumor during measurement, it is possible to reduce the stress on the body of the animal by using two hands. The obtained data include height and topography changes. Figure 7. 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. Discover a faster, simpler path to publishing in a high-quality journal. 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. Determining tumor volume is a useful and quantitative way to monitor tumor progression. 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. (9). Yes 16 (2008). Radiology 254, 110118 (2010). The tumor dimensions are L, W, and H. The volume of an ellipsoid VE, using the variables as shown in Figure 1., is. 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. No, Is the Subject Area "Colorectal cancer" applicable to this article? Traditionally, caliper measurements are used to calculate tumor volume. Mice were killed after 38 days. This process not only deforms the tumor mass but also creates undesirable pain on the animal. For small tumors, the error is about 7%, while for large tumors it is 27%.[3]. . When the tumors became palpable, measurements of tumor volume were performed (with calipers) once per week. Breast Cancer Res. The width of a fully formed tumor will be much larger than its height. NCI CPTC Antibody Characterization Program. The post hoc test resulted in the following. Mammary tumors seemed to take on an oblate spheroid geometry. 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. In Phase IV, the caliper measurement days were the 10th, 12th, 17th, 19th, 22nd and 23rd days. Breast Cancer Res. This does not alter the authors adherence to all the PLOS ONE policies on sharing data and materials. In the case of C38 colon adenocarcinoma, first, the subcutaneously transplanted piece of tumor has to disintegrate, and after that the new tumor colony (which needs to be measured) can begin to grow from the disintegrated tumor cells. For both one has to have the ease of hand and get used to manipulation of the instrument. Similar to the results from the experiments in Figure 18., this error makes the drug seem less effective than it actually is. MeSH Tumor volume determined by microCT, 18 F-FDG-PET and external caliper Tumor volume measured by microCT, PET and caliper all correlated (P < 0.001) with reference volume (figure 1).MicroCT versus reference volume (n = 20) had the best fit of line y = 1.01 0.04x - 6.1 6.3 (R 2 = 0.97; p < 0.001). In Phase III, we have investigated C38 colon adenocarcinoma growth with bevacizumab therapy. Phase III contained three subphases, in every subphases two groups were created; control group received bevacizumab in one dose according to the protocol, while case group received substantially lower doses every day. Research and Innovation Center of Obuda University, Physiological Controls Group, Obuda University, Budapest, Hungary, Affiliation Even though all the three diameters can be measured, the estimation of the volume has quite a large error.