& Med. As it was discussed above, the handling of animals by calipers measurements also suffers such deformations of the animal body during the measurements. 8600 Rockville Pike 47, 510 (2001). Figure 12. Tumor was located in the last third of the back in every cases, thus the effect of respiratory movement was minimal. <> Objectives: Volume measurement of subcutaneous xenograft tumors in nude mice models is an important metric to assess tumor growth or response to therapy. The tumor burden is evaluated by volume calculation using caliper measurement. Please enable it to take advantage of the complete set of features! . Studies using other methods of imaging have exposed the problems that calipers pose. A typical comparison of tumor volume measurements by TumorImager-2TM with calipers. Furthermore, caliper-based measurements cause undue stress on the animal and, by extension, the tumor, which can further distort measurements. Bookshelf Tumor volume (and the third diameter) has to be approximated, assuming a certain shape for the tumor. https://doi.org/10.1371/journal.pone.0142190.g005, https://doi.org/10.1371/journal.pone.0142190.t002, To find the f constant for tumor growth without therapy (Phase I), first reliable tumor volume values had to be found, since there was no MRI measurement in Phase I. Beside tumor diameters, tumor mass was measured and vascularization area was calculated in the case of the removed tumors. J. Urol. 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). Res. 35, 18341840 (2003). Average of tumor volumes for every measurement days of the experiment can be seen in Fig 8. 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). M}{Gt{zj|&cXx[k?CZ ~AjTxMr0 '~ix$mj/)9?$ t-o6MkVXs's}^E6-^_Yrv#w&r7w@WsQs]#G g$Cn. 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. endobj CAS & Moon, R.C. . PubMed Our results for the case group (daily, quasi-continuous small amount administration) and the control group (one big dose according to the protocol) are 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. 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. 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. Circular RNA circTRPS1-2 inhibits the proliferation and migration of esophageal squamous cell carcinoma by reducing the production of ribosomes. In Phase I, 12 mice were implanted with C38 colon adenocarcinoma. We monitored the vital parameters of 4 immunocompetent mice, and there was no serious toxic side-effect or lethality regarding to the usage of bevacizumab. 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. To reduce the time of analysis one could e.g. A caliper only digitizes the length and width of the tumor. 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. 2022 Dec 20;3(12):100852. doi: 10.1016/j.xcrm.2022.100852. In Phase IV, the caliper measurement days were the 10th, 12th, 17th, 19th, 22nd and 23rd days. Statistical evaluations of agreement between two methods for measuring a quantitative variable. Tumor volume was measured with digital caliper as well. ANOVA test showed significant difference between these means (p = 0.041, using 0.05 level of significance). Enter your measurements into the Tumor Volume Calculator to determine the volume of the tumor. Tumor growth was investigated without therapy and with antiangiogenic therapy (using bevacizumab [15]). Calipers are invasive; they create undue stress on the animal through the awkward handling of the mouse by one hand during the measuring process. We compared tumor volumes estimated in vivo by BLI and FLI, to those calculated by caliper measurements and found a good correlation between these two techniques (Figure 4). Correspondence to This device measures the volume displacement of the water due to any object dipped into a measuring cylinder. Cell Rep Med. In Phase IV, the first measurement occurred when the tumors have reached an average volume of 5060 mm3 (according to the xenograft protocol [20]). & Bullock, G.) 253257 (Elsevier Academic, Boston, 2004). 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 . A tumor expands in width rather than height as it grows. Mammary tumors similar to those observed in women can be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea. To overcome all of these difficulties (continuity of the administration and side-effects), model identification should be determined and then a control algorithm (controller) can be designed for the created mathematical model. The mathematical relationship between MRI-measured tumor volume and mass was investigated to estimate accurate tumor volume using caliper-measured data. Tumor was located in the last third of the back of the mouse in every case. 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. Without prior training of users, this effect can lead to greater variability in intra and inter-user measurements. 3.4. Eur. 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. Under these conditions, the formula for the volume of a tumor becomes. Evaluation of prostate volume by transabdominal ultrasonography with modified ellipsoid formula at different stages of benign prostatic hyperplasia. Cancer 83, 621628 (1998). 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. First of all, our hypothesis according to the importance of administration is proved. PASW Statistics 18 (SPSS Statistics, IBM, USA) and Matlab R2009b (MathWorks, USA) were used for statistical analysis. In addition, studies using CT scans concluded that CT yielded tumor volume values 11 to 12% less (an underestimate) than those which were measured by calipers, and that CT-based measurements yielded more accurate and consistent values than caliper-based measurements[6, 7]. The manipulation of the handheld tool is at least as easy as holding a measurement caliper. 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. 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. The most accurate volume calculations were obtained using the formula V = (W2 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 depth. In vivo measurements of tumor metabolism and growth after administration of enzastaurin using small animal FDG positron emission tomography. Kim HB, Zeng CH, Kim Y, Jeong S, Kim SH, Kang JM, Park Y, Won DS, Kim JW, Ryu DS, Lim B, Park JH. & Morel, M. Clinical, mammographic, and sonographic determination of preoperative breast cancer size. ISSN 1548-4475 (online) This is a preview of subscription content, access via your institution. Bevacizumab is an exogenous inhibitor which inhibits the biological activity of human VEGF. Epub 2014 May 28. In Phase IV, case1 group members received 200 g bevacizumab (with 200 l 0.9% NaCl solution) in one dose intraperitoneally on the 8th day. This means that mice which were treated with the recommended bevacizumab protocol (one 200 g bevacizumab dose for an 18-day therapy) did not have significantly smaller tumor volume than mice which did not receive therapy at all. Treat. Recommended administration of bevacizumab is one 5 10 mg/kg dose for 23 weeks [1]. Breast Cancer Res. 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. Powered by WordPress In Phase III/3, case group members received one-tenth dose of control dose intraperitoneally spread over 18 days. Cells should be suspended in a volume so that 300 l contains required number of cells per injection. Modern Pathol. Epub 2017 Mar 21. A tumor image, its 3D shape and all geometrical data are automatically calculated and stored. Figure 17. Kubatka, P. et al. A possible solution for height approximation [23] is: Bethesda, MD 20894, Web Policies Because the industrys caliper-based formula allows for these inclusions in nonexistent volumes more often than not, it yields inaccurate measurements, which can confuse. The results of the three investigated cases using C38 mouse colon adenocarcinoma (Phase I, Phase III/3 control group, and Phase III/3 case group) were compared using tumor volume values from MRI measurements (Phase III/3 control and case groups, 23rd day) and evaluated data (Phase I MRI tumor volume calculated from linear curve fit, 24th day). Phase I and Phase III/3 control groups are not significantly different (p = 0.572), while Phase I and Phase III/3 control groups are significantly different (p = 0.002). (1). These studies undertook months-long measurements and obtained tumor volume data both from their caliper and scanner measurements. & Robinson, S.P. Mrio Ginja. 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). Isofluorane (0.95 x 2.0%) was applied for inhalational anesthesia, and intubation was performed. 2017 May 1;176:35-41. doi: 10.1016/j.lfs.2017.03.013. 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. Breast Cancer Res. If this surface intersects above the bottom of the tumor, then the volume estimation of the tumor will be smaller than the true value as can be seen from Figure 9. They found that caliper and ultrasonography measurements were significantly correlated but that tumor volumes calculated using different formulas varied substantially. Pursuant to one-sample Kolmogorov-Smirnov test, each sample has normal distribution (ppIV_control = 0.981, ppIV_case1 = 0.998, ppIV_case2 = 0.854). bowel, and kidneys is when the tumor is located in . b) Fluorescence picture was created using CD31 antibody immunohistochemistry staining to calculate vascularization area. (6) (7). Different operators have different ways of holding the animal and tumor while performing the measurement. Possible mechanisms mediating an association between physical activity and breast cancer. Denis, F., Paon, L. & Tranquart, F. Radiosensitivity of rat mammary tumors correlates with early vessel changes assessed by Power Doppler sonography. Finding the mathematical relationship between MRI-measured tumor volume and tumor mass creates the possibility to estimate tumor volume from caliper-measured data. Mice were monitored daily by personnel experienced in recognizing signs of morbidity (illness, injury, or abnormal behavior) also in weekends and holidays. 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. J. Oncol. Weber, W.A. Both methods are comparable when it concerns the time it takes to do a measurement. https://doi.org/10.1371/journal.pone.0142190.g004. {(Q~J9}=ow[5nkW"C; >tDve+7K,}g?A Berg, W.A. 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 15. Cancer Inst. and JavaScript. Cancer Res. These images were applied to calculate vascularization area by using ImageJ (NIH, USA) [33] software (Fig 4). One common measure of efficacy is the T/C ratio, the ratio of tumor volume in control versus treated mice at a specified 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. This does not alter the authors adherence to all the PLOS ONE policies on sharing data and materials. Department of Control Engineering and Information Technology, Budapest University of Technology and Economics, Budapest, Hungary, Affiliation J. Natl. sharing sensitive information, make sure youre on a federal 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. Figure 6. A study[4]on caliper measurements, shoved the very poor accuracy of the formula used for caliper measurements. This creates the possibility of further efficient therapeutic agent use (if any) or assists the more effective direct antitumoral effect of bevacizumab. Experimentally induced mammary tumors in rats. This results in the inclusion of sections of the parallelepipeds volume that does not exist in the actual tumor. Mammary tumors similar to those observed in women can be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea. (8) Using the above mentioned iterative method, the resulted equation is Automatic randomization to perform group assignments based on body weight, tumor area/volume, delta tumor volume, or any numeric value and/or user-defined formula; Make sure you get accurate measurements for length, width and height. In Phase IV, control group members received no bevacizumab. J. Clin. Examination of tumor growth belongs not only to the basic medical research, but to the biomedical engineering field as well. Yes & Hafstrm, L. Estimation of liver tumor volume using different formulasan experimental study in rats. Fj^P;7? 2001 Jan 15;19(2):551-7. doi: 10.1200/JCO.2001.19.2.551. 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. On Day 14, when the tumor volume reached ~500 mm 3, the mice were . A two-dimensional mathematical model was created from the experimental results of BALB/c mice with KHJJ tumor line [25]: Tumors should also be placed so that the tumor peak is near the center of the mask and is not highly inclined. The majority of debate over Avastin is about breast cancer because, in 2011, the US Food and Drug Administration (FDA) revoked the approval of Avastin for breast cancer treatment in the absence of decisive therapeutic benefit; however, several clinical trials suggested that Avastin can be effective in breast cancer treatment [6, 7]. 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 volume was measured with digital caliper. Mice were humanely sacrificed using cervical dislocation. Care should be taken not to stretch the mouse whether using one or two hands to hold the mouse during a scan. One can find datasets in Table 3. The post hoc test resulted in the following. 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. The lines on the graphs have a slope of 1, signifying that it represents caliper-based measurements in which height equals width. Provided by the Springer Nature SharedIt content-sharing initiative, Bulletin of the National Research Centre (2022), Lab Animal (Lab Anim) Tumor volumes have been calculated for all A431 tumors by the conventional caliper/formula method and by ultrasound with the ARTIDA system. For small tumors, the error is about 7%, while for large tumors it is 27%.[3]. Current standard technique for volume determination of subcutaneously xenografted tumors in vivo is by external caliper where tumor volume is calculated by use of the modified ellipsoid formula 1/2(Length Width 2)[1,2]. All surgery and sacrifice were performed under sodium pentobarbital anaesthesia (Nembutal, 70 mg/kg). CAS A two-dimensional mathematical model was applied for tumor volume evaluation, and tumor- and therapy-specific constants were calculated for the three different groups. Mammary tumors seemed to take on an oblate spheroid geometry. In Phase IV, approximately 2.9106 cells were injected subcutaneously into each SCID mouse. Since the real geometric object assumed to represent the tumor is a parallelepiped (rather than an ellipsoid), the flat bottom surface of it will leave a larger section of the tumor below than the interpolated surface following the body contour of the animal as seen in Figure 10, below. In that caseeven though all the three diameters can be measuredthe estimation of the volume has quite a large error. Tumor Volume Formula for Caliper Measurements, 3. 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. 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. 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 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. Comparing ARTIDA-calculated tumor volumes to the volumes derived from the caliper/ellipsoid formula calculations and referenced to the gold standard volume determination method (with literature density values), as it is shown in Figure 2A. J. Nucl. . 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. PLoS ONE 10(11): How the device is positioned over the tumor will not influence the result. A direct and real 3D volume of the xenograft will then be calculated. To validate our results which come from the experiments where mouse tumor vascularization was inhibited with humanized VEGF, we evaluated the results of Phase IV, where immunocompromized (SCID) mice were used with human colorectal adenocarcinoma (HT-29) xenografts. 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