For the TM900 one has to push the acquisition button and almost immediately the images are stored and values calculated. 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]. 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). Eichelerger, L.E. Click through the PLOS taxonomy to find articles in your field. & trukelj, B. N-methylnitrosourea induced breast cancer in rat, the histopathology of the resulting tumours and its drawbacks as a model. Furthermore, caliper-based measurements cause undue stress on the animal and, by extension, the tumor, which can further distort measurements. 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. 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]). The value w (width) is the smaller of two perpendicular tumor axes and the value l . Westerlind, K.C. Tumor as a 3-D object has three diameters (width, length and height); however in vivo only two diameters can be measured by caliper (width, w and length, l). According to the Xenograft tumor model protocol [20], tumor volume has to be calculated using the following formula: In this case, tumor volume has to be calculated in the following way: 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 . As it was discussed above, the handling of animals by calipers measurements also suffers such deformations of the animal body during the measurements. 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. To reduce the time of analysis one could e.g. To obtain 15, 115121 (2009). The system includes database, data management and processing software and can be complemented with a weight . Tumor volume measurement was performed by digital caliper and small animal MRI. We found the same result for human colorectal adenocarcinoma (HT-29) xenografts. For both one has to have the ease of hand and get used to manipulation of the instrument. Determining tumor volume is a useful and quantitative way to monitor tumor progression. It was on the 10th day, since HT-29 colorectal adenocarcinoma grows slower in mice than C38 colon adenocarcinoma. Notice that the caliper-based measurements have a larger standard deviation. (4). Our hypothesis is that the effectiveness of a lower dosage with a quasi-continuous therapy can be comparable with the protocol therapy. & Pisani, P. Global cancer statistics, 2002. Measurements of tumors, in the context of research and industry, are expected to be accurate; it is unacceptable that this inaccurate formula has stood so long as the industry standard. Clin. Table 1. 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. Additionally, this technique is plagued by the inconsistent selection of tumor axes at subsequent examinations. How the device is positioned over the tumor will not influence the result. A smaller standard deviation in a set of measurements suggests a lesser possibility of error. If the length or width of a tumor is twice its height, which is often the case for many tumor models, then its measured volume will be overestimated by 50%. Breathing was monitored with piezoelectric transducer; temperature of the body was measured by rectal thermometer. Faustino-Rocha, A., Oliveira, P., Pinho-Oliveira, J. et al. The results presented here were provided by them. 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. No mice died during the experiment, therefore 5 mice from the control group, and 9 mice from the case group were sacrificed on the 23rd day. 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. %PDF-1.4 That is independent from the use of a direct optical 3D measurement tool or a caliper. 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). The overestimation of caliper measurements is due mainly to the fact that the formula used is that of a parallelepiped, a shape unlike an ellipsoid actually represents the geometric body that is being measured. Measured data is the MRI-measured tumor volumetumor mass pairs on the 23rd day of Phase III/3 (case and control group). 35, 18341840 (2003). Analyzed the data: JS ZS PK DG DAD LK. The C38 colorectal carcinoma line was maintained by serial subcutaneous transplantations in C57Bl/6 mice. 55, 74108 (2005). 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. In this study, the authors measured dimensions of rat mammary tumors using a caliper and using real-time compound B-mode ultrasonography. In Phase I, tumor growth was investigated without antiangiogenic therapy for 24 days (mice were sacrificed when the tumor reached a lethal size). Evaluation of prostate volume by transabdominal ultrasonography with modified ellipsoid formula at different stages of benign prostatic hyperplasia. Biol. Description of Lesions. Tumor volumes ranged from 0.5 cc to 187.9 cc, with a median of 21.8 cc. Affiliation 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. Most of the operators in these studies were highly experienced in caliper measurements, but they only had one day of training in scanner measurements. Tumor volume was measured in two different ways. n5). The aim of the experiment was to create and validate a clinically relevant tumor growth model (using C38 colon adenocarcinoma), focusing on the effect of angiogenesis. 19, 6170 (1989). Weber, W.A. The resulted linear curve is Jensen, M.M., Jrgensen, J.T., Binderup, T. & Kjr, A. Tumor volume in subcutaneous mouse xenografts measured by microCT is more accurate and reproducible than determined by 18F-FDG-microPET or external caliper. Quantity of the optimal solvent administration was also examined in this subphase. Comparison of clinical assessment, mammography and ultrasound in pre-operative estimation of primary breast-cancer size: a practical approach. Catheter was placed in the tail vein for injectionaccording to the mouse tail vein injection protocol [30]to investigate drug effect. . to calculate several estimates of tumor volume using the equations for the volume of a sphere ([4 /3]r3), Here r is average of a, b and c the volume of an Figure 5. The closer the slope of a line on this graph is to 1 and the y-intercept to 0, the closer the measurements of the device it represents are to the exact values of the tumors volumes. and calculates an interpolated surface under the tumor boundary based on the healthy surface around the tumor (navy blue line in Figure 12.). In this phase, 5 immunocompetent mice in control group, and 9 immunocompetent mice in case group were used. 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. & Wieder, H. Monitoring chemotherapy and radiotherapy of solid tumors. Accuracy of Optical Imaging for Measuring Tumor Burden In Vivo . Med. Of course, for the closed-loop design frequent and precise tumor volume measuremets are required, thus the problem of tumor volume measurement has to be solved as well. These model-based protocols can be more effective than the current ones, since they provide individual treatment for the patients. . & Russo, I.H. The TM900 also stores an optical image in the measurement data base. The study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. Normality was investigated with one-sample Kolmogorov-Smirnov test, and homogeneity of variance (homoscedasticity) was examined with Levenes test. 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. When using calipers there is dependency in the measured data of the positioning by the operator of the tool on the xenograft. We compared the volumes, areas . In longitudinal studies, sequential measurements of tumor volume with a non-invasive method are essential. Using the mathematical model, different dosage algorithms can be designed for antiangiogenic cancer therapy [1619]. All surgery and sacrifice were performed under sodium pentobarbital anaesthesia (Nembutal, 70 mg/kg). 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. In order to assess interobserver variation, each . Preclinical evaluation of LU 79553: a novel bis-naphthalimide with potent antitumor activity. & Robinson, S.P. 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. Additionally, we explore the impact of six different tumour . Lab Animal Sport. In addition, the root mean square, R2 values under the linear regression equations in Figure 15. symbolize the precision of the data. 2015 Feb;43(2):145-70. doi: 10.1177/0192623314532036. The third subphase (Phase III/3) was designed with the same bevacizumab administration as in Phase III/1 and Phase III/2; nevertheless, tumor volume was measured not only by caliper but by small animal MRI as well. For more information about PLOS Subject Areas, click Using an extremely low dosage (as in our study), there is a high probability that these adverse effects can be minimized. 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. 29 September 2022, Bulletin of Mathematical Biology Further geometrical or visual data are not stored simultaneously. Figure 2. 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary. 2 0 obj volume42,pages 217224 (2013)Cite this article. Pairwise comparison was done by Tukeys honest significant difference test to find those samples, which have significantly different means. A standard deviation of 0% is almost impossible in real life, but a smaller standard deviation is generally favorable. An average penny has a diameter of 19 mm and a height of 1.27 mm, yielding a volume of 360.1 mm3. In one trial, 208 tumors were measured, and 98.94% of those tumors exhibited much smaller heights than widths. Google Scholar. 47, 510 (2001). 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). Another approach is to utilize the scan images (Figure 14.) Lifetime dose-response relationships for mammary tumor induction by a single administration of N-methyl-N-nitrosourea. 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. In this way, the targeted therapy can be used in a much more individualized form. Without prior training of users, this effect can lead to greater variability in intra and inter-user measurements. (3), Another relatively new but not widely used approach to estimate tumor volume is to assume hemi-ellipsoid shape [24]. Kubatka, P. et al. This means that continuous, low dose antiangiogenic factor (VEGF) administration is needed. Clarke, R. Animal models of breast cancer: their diversity and role in biomedical research. In Phase IV, the first measurement occurred when the tumors have reached an average volume of 5060 mm3 (according to the xenograft protocol [20]). Epub 2014 May 28. Tumor was located in the last third of the back in every cases, thus the effect of respiratory movement was minimal. Tumor segmentation by TumorImager-2TM. Tumor growth inhibition rate (%); can be calculated using the following formula: = (Vc 1 -Vt 1 )/ (Vc 0 -Vt 0) X 100. where, Vc 1 is the mean tumor volume in the control group at the time of tumor . 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. Mammary tumors seemed to take on an oblate spheroid geometry. a) Haematoxylin Eosin (H&E) staining was applied to investigate tumor morphology. 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. 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. A diagram of an ellipsoid, with the derivations of the variables needed to calculate its volume. Tumor growth was investigated without therapy and with antiangiogenic therapy (using bevacizumab [15]). Harris, R.E., Alshafie, G.A., Abou-Issa, H. & Seibert, K. Chemopreventive of breast cancer in rats by celecoxib, a ciclooxygenase 2 inhibitor. Better results can be obtained by proper handling of the mice. 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. 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). Tumor volume was calculated by the modified ellipsoidal formula: V = (Length Width 2) . 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. where f is a constant which belongs to a certain tumor type. Tumor growth Tumor regression Number of Number of volume volume doublings Percentage additional halvings: Decrease in Percentage decrcase in increase Volume Area" Diameterb volume doubling number Volume Area" Diameterb Doubling of Halving of Volume 1 .o 100.0 58.7 26.0 Volume 1 .o 50.0 37.0 20.6 (Budapest, Hungary). In both cases, three groups were compared in the experiments. A two-dimensional mathematical model was created from the experimental results of BALB/c mice with KHJJ tumor line [25]: <> 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. Figure 17. Yes The article discusses two results. 16 (2008). PubMedGoogle Scholar. 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. However, most serious questions are: for how long and continuous or not? Fornage, B.D., Toubas, O. Tumor volumes are calculated from digital caliper raw data by using the formula:Volume (mm3) = (l x w2) / 2. 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. calculate an average volume based on two extreme segmentations; one including most of the border, and one excluding it. Frozen samples were used to create 15 m frozen cuts. Accessibility Med. This device measures the volume displacement of the water due to any object dipped into a measuring cylinder. 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. It means that a mouse of the case group received 1.11 g bevacizumab (with 45 l 0.9% NaCl solution) every day from the 3rd day for 20 days. Therefore, optical imaging is . Girit, I.C., Jure-Kunkel, M. & McIntyre, K.W. For the sake of convenience, establishments will simplify and H. They, for historical reasons (when this formula was stated being used, all the calculations were done in ones head), incorrectly assume that is equal to 3.0 and that the height of a tumor (H) is equal to its width (W). In Phase IV, control group members received no bevacizumab. 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. Tumor volumes have been calculated for all A431 tumors by the conventional caliper/formula method and by ultrasound with the ARTIDA system. First of all, our hypothesis according to the importance of administration is proved. 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. Bookshelf Conceived and designed the experiments: ZS JS. 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E ) staining was applied to investigate tumor morphology cancer treatment '' applicable to this article caliper-based measurements cause stress. Phase, 5 immunocompetent mice in control group ) approach is to assume hemi-ellipsoid [. Animal body during the measurements to any object dipped into a Measuring cylinder designed the experiments: ZS JS test... Calculated for all A431 tumors by the Institutional animal Care and use Committees protocols ( )... Pubmed tumors this large are typically beyond the maximum size allowed by the Institutional animal Care use... The same result for human colorectal adenocarcinoma ( HT-29 ) xenografts ZS PK DAD... Software and can be obtained by proper handling of the water due to any object dipped into a cylinder. Body was measured by rectal thermometer the ARTIDA system tumors by the modified ellipsoidal formula: =., Budapest, Hungary is to utilize the scan images ( Figure 14. thermometer... 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