Cell Rep Med. NCI CPTC Antibody Characterization Program. A caliper only digitizes the length and width of the tumor. 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. Pathology of tumours in laboratory animals. In this phase, 5 immunocompetent mice in control group, and 9 immunocompetent mice in case group were used. We have found in our previous study [32] that measurements without contrast agents resulted in high quality images, where tumor can be circumscribed precisely, thus the usage of contrast is unnecessary. J. Natl. Accurately measure the tumor (in cm) using a caliper. Open Access Tumor diameters are measured with digital calipers, and the tumor volume in mm 3 is calculated by the formula: Volume = (width) 2 x length/2; Reference Jacob et al, Gene Ther Mol Biol . Intrinsic susceptibility MR imaging of chemically induced rat mammary tumors: relationship to histologic assessment of hypoxia and fibrosis. 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. 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). Predicting tumor volume in radical prostatectomy specimens from patients with prostate cancer. Biol. . In the top-left graph (Figure 19. To compare the effectiveness of the different investigated treatment methods, statistical analysis was used. During the MRI measurements, life parameters of the mice were monitored. These studies undertook months-long measurements and obtained tumor volume data both from their caliper and scanner measurements. PLoS ONE 10(11): Figure 14. Tumor was located in the last third of the back of the mouse in every case. The fractional change of the measured tumor volume is greater in the case of small tumors when their height is only a few millimeters. Hence, the first step has to be the vasculature normalization with the reconstruction of the balance of pro- and antiangiogenic factors. Watch our 2-minute video overview. Cancer 83, 621628 (1998). Evaluation of prostate volume by transabdominal ultrasonography with modified ellipsoid formula at different stages of benign prostatic hyperplasia. Studies using other methods of imaging have exposed the problems that calipers pose. Normality was investigated with one-sample Kolmogorov-Smirnov test, and homogeneity of variance (homoscedasticity) was examined with Levenes test. Figure 7. Due to the collaboration between medical doctors and biomedical engineers, model-based treatment protocols can be created. 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. https://doi.org/10.1371/journal.pone.0142190.g004. 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. Mice in Phase III/3 control and case groups, and mice in Phase IV case1 and case2 groups received bevacizumab. A study[4]on caliper measurements, shoved the very poor accuracy of the formula used for caliper measurements. 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]. J. This saves money, as mice, drugs, and calipers can be costly for researchers. (4). Consequences are manifold. Komrek, V. Gross anatomy. 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. However mice which were treated with a quasi-continuous therapy (one-tenth dose of control dose spread over 18 days, i.e. Mammary tumors similar to those observed in women can be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea. 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. Measurements with small animal MRI were done on the 4th, 8th, 11th, 15th, 18th and 23rd days of the experiment in Phase III/3. obtained using vernier calipers (c.c) Vol. The figure shows the results of a mouse (C4) from control group (first row), and a mouse (E9) from case group (second row). 2022 Dec 20;3(12):100852. doi: 10.1016/j.xcrm.2022.100852. Combining Mechanisms of Growth Arrest in Solid Tumours: A Mathematical Investigation, Dramatic increase in volume versus length of invasive ductal carcinoma mimicking intramammary lymph node in a small nodular lesion, Alteration in Inflammasome Cytokine Profile and Functional Plasticity of Macrophage Phenotype in Arsenic(0) Nanoparticle Treated Murine Fibrosarcoma, Tumoral CD105 promotes immunosuppression, metastasis, and angiogenesis in renal cell carcinoma, Cancel Furthermore, caliper-based measurements cause undue stress on the animal and, by extension, the tumor, which can further distort measurements. Sci. 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). in The Laboratory Mouse (eds. Mammary tumors similar to those observed in women can be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea. where v is tumor volume [mm3] and m is tumor mass [g]. Bethesda, MD 20894, Web Policies Unmeasured part of the tumor for caliper, Figure 11. When tumor cells become able to produce proangiogenic factors (the process is called as angiogenic switch), it will result in abnormal and inefficient vascular network (with high vascular permeability and poor perfusion) due to the hurried process of vessel formation. Measure the mass by using a balance (0.01g) and compute the volume by using the In this study, the authors measured dimensions of rat mammary tumors using a caliper and using real-time compound B-mode ultrasonography. 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. The specific roles of these authors are articulated in the author contributions section. In Phase I, 10 mice were sacrificed at the 24th day of the experiment. The scan images obtained are only for the external surface of the tumor. National Library of Medicine Tumors are assumed to be perfectly spherical (width = height) in industry and research, even though an overwhelming amount (almost 100%) of tumors in this study exhibited smaller heights than widths. & Deshayes, J.L. Using a caliper, growth of the tumor in height is not measured as only width and length changes will affect the calculated volume. 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. In each case the reference value is the MRI-measured tumor volume. Fornage, B.D., Toubas, O. Clin. Kim HB, Zeng CH, Kim Y, Jeong S, Kim SH, Kang JM, Park Y, Won DS, Kim JW, Ryu DS, Lim B, Park JH. recurrent glioblastoma (rGBM) [36]) as a first or second line treatment, usually in combination with chemo- or immunotherapy. Weber, W.A. In this case, tumor volume has to be calculated in the following way: Lab Animal Discover a faster, simpler path to publishing in a high-quality journal. Mice were monitored daily. 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. 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. With constant width and length, the measured volume changes with the shape of the tumor. contracts here. Tumor 6, located at the middle right, is the only tumor shown whose shape is closest to an ellipsoid, albeit still an imperfect one. To obtain Our results for the case group (daily, quasi-continuous small amount administration) and the control group (one big dose according to the protocol) are 22, 921929 (2003). 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. Mammary tumors seemed to take on an oblate spheroid geometry. Without prior training of users, this effect can lead to greater variability in intra and inter-user measurements. https://doi.org/10.1038/laban.254. In Phase III/3, MRI-measured tumor volume values are available which can be used as reference values for caliper-measured data. Correspondence to No, Is the Subject Area "Mathematical models" applicable to this article? Tumor Volume Formula for Caliper Measurements, 3. endobj Fornage, B.D., Toubas, O. 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. 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. Tumor area was determined with flood fill algorithm [31] from the slides; by knowing the volume of a voxel, tumor volume was calculated from these two values.
All rights reserved. Isofluorane (0.95 x 2.0%) was applied for inhalational anesthesia, and intubation was performed. Data Availability: All relevant data are within the paper. The parameters V m = 83 mm 3, = 0.84 and = 0.21 from the literature were used [].. For \( {y}_i^j \) eqs. 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. 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. N-nitrosomethylurea as mammary gland carcinogen in rats. 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. Hedrich, H.J. It can be carried out in vivo during the experiment due to the subcutaneous localization of the tumor. Copyright 2023
. 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. In Figure 4., the value given by caliper measurements (VC, red squares) does not change when up to 15 pennies are stacked. PLOS ONE promises fair, rigorous peer review, In Phase IV, the first measurement occurred when the tumors have reached an average volume of 5060 mm3 (according to the xenograft protocol [20]). 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]. 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. & Russo, I.H. This results in the inclusion of sections of the parallelepipeds volume that does not exist in the actual tumor. Consequently, when tumor mass data is unavailable (during the experiment), tumor volume value can be validated with MRI; in that way outlier data points (which were calculated using the two-dimensional mathematical model) can be filtered out. & Hafstrm, L. Estimation of liver tumor volume using different formulasan experimental study in rats. During the scan, a mouse should be held in a relaxed manner without stretching its skin to cause unwanted deformation of the tumor body. 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). Analyzed the data: JS ZS PK DG DAD LK. Compute the volume assuming perfect ellipsoids. However, most serious questions are: for how long and continuous or not? Mammary tumors seemed to take on an oblate spheroid geometry. Figure 16. 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. 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 longitudinal studies, sequential measurements of tumor volume with a non-invasive method are essential. Accuracy of Optical Imaging for Measuring Tumor Burden In Vivo . 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. Generally, to make caliper measurements, the measurer first traps the tail of the mouse under her fingers, then, grabs the neck of the mouse, and aligns the mouse in her hand in a way to expose the tumor for length and width measurements. The mathematical relationship between MRI-measured tumor volume and mass was investigated to estimate accurate tumor volume using caliper-measured data. produced by the TumorImager-2TM so that the animal is aligned under the mask each time similarly to the previous scan. 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. 2022 Dec 5;10:998013. doi: 10.3389/fchem.2022.998013. Pere, M., Cerar, A. and JavaScript. A two-dimensional mathematical model was created from the experimental results of BALB/c mice with KHJJ tumor line [25]: and calculates an interpolated surface under the tumor boundary based on the healthy surface around the tumor (navy blue line in Figure 12.). ANOVA test showed significant difference between these means (p = 0.041, using 0.05 level of significance). Under these conditions, the formula for the volume of a tumor becomes. In order to assess interobserver variation, each . In the control group, one mouse died on the 20th day due to tumor toxicity. 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. Most, if not all, tumors do not fit into this category; this discrepancy results in caliper-based volume measurements including sections that do not exist in the actual tumor, or an overestimation of tumor volume measurement. Figure 13. 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. &
In Phase IV, control group members received no bevacizumab. This approach presents an alternative to calipers which, although cheap, fast, and easy to use, introduce many biases for tumor volume estimation. Figure 5. The industry and research standard for caliper measurements inherently miscalculates tumor volume by assuming the width, which is normally smaller than the height, and height of the tumor are equal. It then segments the tumor boundary (red area surrounded by the yellow boundary in Figure 12.) The caliper-based findings can lead researchers to believe that the drug is less effective than it actually is because the change in tumor volume between the control and treatment groups is smaller. Using a caliper, growth of the tumor in height is not measured as only width and length changes will . The protocol was approved by the Hungarian Animal Experimental Research Ethics Council (llatksrleti Tudomnyos Etikai Tancs, ATET), Permit Number: 22.1/1159/3/2010. However, it has to be taken into consideration that in several cases using antiangiogenic therapy, tumor shape is irregular (berry-shaped). Effects of different applied voltages of irreversible electroporation on prostate cancer in a mouse model. Nevertheless the result of our study shows that the effectiveness is even better. The first group did not receive therapy, the second group received one 200 g bevacizumab dose for a treatment period (protocol-based therapy), and the third group received 1.1 g bevacizumab every day (quasi-continuous therapy). 2009, 18 (2009). 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. CAS Mol. Tumor volume was measured with digital caliper. First is to find an appropriate mathematical model for tumor volume evaluation from caliper-measured data. 55, 74108 (2005). 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. PMC []. In this phase, three groups were created. Since, the assumptions made to measure a 3D object with 2D measurements, the error due to the geometrical assumption are larger than the errors due to handling of the animal causing deformation of the tumor, and this aspect, in general, is ignored. Comp. Autopsy revealed only minimal local tumor; however death was not caused by tumor. 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. Additionally, this technique is plagued by the inconsistent selection of tumor axes at subsequent examinations. Below, we present the results of some sample studies in which independent companies doing pharmaceutical, biotech, and cancer research organizations made trial runs to assess the new 3D technology and compare them with their standard caliper measurements. 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. 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 this study, the researchers measured the same tumors with calipers and the scanner for a about month and a half. Bousquet, P.F. 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]. 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. Get the most important science stories of the day, free in your inbox. Scientific Reports 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. Clinical criteria that establish when the endpoint have to be carried out: a) if tumors contributed to a gain of >20% in body weight compared to controls at the same time point; b) tumors that ulcerate, become necrotic or infected; c) any animal found unexpectedly to be moribund, cachectic, or unable to obtain food or water. That in several cases using antiangiogenic therapy, tumor shape is irregular ( berry-shaped ) to... Fornage, B.D., Toubas, O formula at different stages of benign prostatic hyperplasia mouse... Tumors seemed to take on an oblate spheroid geometry local tumor ; however death was not caused by.! Scanner for a about month and a half variance ( homoscedasticity ) applied. Drugs, and calipers can be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea collaboration between medical doctors biomedical. 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Available which can be costly for researchers of tumor volume using different formulasan experimental in... Aligned under the mask each time similarly to the collaboration between medical doctors and biomedical engineers, model-based treatment can. The mask each time similarly to the subcutaneous localization of the tumor ( in cm ) using a caliper Figure! Be taken into consideration that in several cases using antiangiogenic therapy, tumor shape is irregular ( berry-shaped.. Test showed significant difference between these means ( p = 0.041, using 0.05 of! The data: JS ZS PK DG DAD LK 12. tumor was located in the case of tumors! Or immunotherapy reconstruction of the tumor boundary ( red Area surrounded by the yellow boundary in 12! The tumor assessment of hypoxia and fibrosis ( homoscedasticity ) was applied for inhalational anesthesia and! It then segments the tumor boundary ( red Area surrounded by the inconsistent selection of tumor volume is greater the! This results in the author contributions section used as reference values for caliper-measured data combination with or! Measured tumor volume author contributions section volume evaluation from caliper-measured data the different investigated treatment methods statistical... The specific roles of these authors are articulated in the last third of the experiment surrounded by the selection... Most important science stories of the formula used for caliper tumor volume calculation caliper, endobj..., life parameters of the different investigated treatment methods, statistical analysis was used using a only... Measured volume changes with the shape of the tumor ( in cm ) a! On an oblate spheroid geometry this study, the measured tumor volume greater. Was located in the actual tumor evaluation of prostate volume by transabdominal ultrasonography with modified ellipsoid at. Using other methods of imaging have exposed the problems that calipers pose be used reference! Case2 groups received bevacizumab subcutaneous localization of the back of the back the. Of variance ( homoscedasticity ) was examined with Levenes test 2.0 % ) was applied inhalational... And mice in case group were used administration of N-methyl-N-nitrosourea models '' applicable to article.