{(Q~J9}=ow[5nkW"C; >tDve+7K,}g?A Consequences are manifold. The fractional change of the measured tumor volume is greater in the case of small tumors when their height is only a few millimeters. Epub 2017 Mar 21. The mathematical relationship between MRI-measured tumor volume and mass was investigated to estimate accurate tumor volume using caliper-measured data. The closer the shape of the tumor is to a perfect ellipsoid, as in the region of the graph in which H closely W, the more closely caliper measurements mirror real-life measurements. This concept can easily be understood by a simple thought experiment: the volume of a stack of pennies (Figure 4.). Tumor growth was investigated without therapy and with antiangiogenic therapy (using bevacizumab [15]). In the case of HT-29 adenocarcinoma, the daily treatment with one-twelfth total dose resulted in significantly smaller tumors than the protocol-based treatment (p = 0.038). Bizberg Themes, By Memorial Sloan Kettering Cancer Center. The produced images were converted to NIfTI (Neuroimaging Informatics Technology Initiative) format, which is suitable for image processing. Position of the mouse was fixed to minimize the movement of the animal. Parkin, D.M., Bray, F., Ferlay, J. In the top-left graph (Figure 19. 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 Treat. Tumor Volume Measurement In Phase I, Phase II and Phase III, a piece of C38 colon adenocarcinoma was transplanted subcutaneously in the recipient animal on the 1st day of the experiment (approximately 2106 cells were transplanted into each mouse). After sacrificing mice, tumors were removed, and their mass was measured. Using a caliper, growth of the tumor in height is not measured as only width and length changes will affect the calculated volume. Bookshelf 2009, 18 (2009). The caliper, unlike the scanner, does not consider that the height of the stack changes while the length and width remain the same. Bevacizumab administration was started on the 7th day in both cases. 1 0 obj Analyzed the data: JS ZS PK DG DAD LK. & Hafstrm, L. Estimation of liver tumor volume using different formulasan experimental study in rats. 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary. . Compute the volume assuming perfect ellipsoids. where V m is the measurement threshold that represents the smallest measurable tumor volume using a caliper. All mice were implanted with HT-29 human colorectal adenocarcinoma. In Phase IV, the caliper measurement days were the 10th, 12th, 17th, 19th, 22nd and 23rd days. 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 The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Even the same operator, with different trials or at different moments in time does not always hold the animal in exactly the same way. Tumor was located in the last third of the back in every cases, thus the effect of respiratory movement was minimal. Furthermore, caliper-based measurements cause undue stress on the animal and, by extension, the tumor, which can further distort measurements. Estimation of rat mammary tumor volume using caliper and ultrasonography measurements. Copyright 2023 Powered by WordPress This effect is illustrated in the included video. Radiology 254, 110118 (2010). This means that the daily treatment with one-twelfth total dose resulted in significantly smaller tumors than the protocol-based treatment. Another approach is to utilize the scan images (Figure 14.) 60, 21012103 (2000). With a caliper measurement, the angle at which the tool is held over the tumor will affect the obtained value, thus inducing an extra operator and manipulation sensitivity in the final results. Med. official website and that any information you provide is encrypted Open Access articles citing this article. mum tumor volume of 0.6 mL was reached. Bevacizumab (Avastin) [1] is an exogenous inhibitor, which inhibits the biological activity of human VEGF [2]. 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 best experience, we recommend you use a more up to date browser (or turn off compatibility mode in and calculates an interpolated surface under the tumor boundary based on the healthy surface around the tumor (navy blue line in Figure 12.). These images were applied to calculate vascularization area by using ImageJ (NIH, USA) [33] software (Fig 4). Careers. This surface has x, y, and z (equivalent to L, W, and H of caliper measurements for every point on the surface) components. 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. Healthy tissues have efficient vasculature and effective blood supply due to the balance of proangiogenic and antiangiogenic factors. 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. On the bottom left graph in Fig. In this study, the researchers measured the same tumors with calipers and the scanner for a about month and a half. Girit, I.C., Jure-Kunkel, M. & McIntyre, K.W. Clarke, R. Animal models of breast cancer: their diversity and role in biomedical research. On Day 14, when the tumor volume reached ~500 mm 3, the mice were . Inhibiting the glycerophosphodiesterase EDI3 in ER-HER2+ breast cancer cells resistant to HER2-targeted therapy reduces viability and tumour growth. J. Ultrasound Med. All surgery and sacrifice were performed under sodium pentobarbital anaesthesia (Nembutal, 70 mg/kg), and all efforts were made to minimize suffering. government site. Bevacizumab is a humanized monoclonal antibody, therefore the effectivity of bevacizumab therapy in mice is not trivial. No animals were sacrificed before the experimental endpoint. In Phase I, we have investigated tumor growth without therapy with two types of mouse tumor. N-nitrosomethylurea as mammary gland carcinogen in rats. In Phase IV, approximately 2.9106 cells were injected subcutaneously into each SCID mouse. 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. 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. Figure 13. ANOVA test showed significant difference between these means (p = 0.041, using 0.05 level of significance). This creates the possibility of further efficient therapeutic agent use (if any) or assists the more effective direct antitumoral effect of bevacizumab. These measurements gave us accurate tumor volumes to compare the accuracy of this new technique in measuring tumor volumes. Tievsky AL, Ziegler RB, Salhus MR, Weisskoff R. Comparison of diameter and perimeter methods for tumor volume calculation. Numerical result can be found in Table 2, and Fig 5 presents graphical results. The lines on the graphs have a slope of 1, signifying that it represents caliper-based measurements in which height equals width. Mice were killed after 38 days. CAS Figure 2. 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. et al. where v is tumor volume [mm3] and m is tumor mass [g]. McCormick, D.L., Adamowski, C.B., Fiks, 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. 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. 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. 105, 2023 (1983). 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. Additionally, this technique is plagued by the inconsistent selection of tumor axes at subsequent examinations. Tumours of the rat. 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. Materials and methods: The murine CaNT subcutaneous xenograft model was used. Med. & 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. J. Possible mechanisms mediating an association between physical activity and breast cancer. 12, 881885 (2005). 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. Our study of the incised tumors in many mouse tumor models showed that this is mostly seen for very large tumors (Volume > ~1.5 cm3) where the tumors start bulging inward toward the flesh. Faustino-Rocha AI, Gama A, Oliveira PA, Vanderperren K, Saunders JH, Pires MJ, Ferreira R, Ginja M. Life Sci. 17, 905910 (2004). 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. Tukeys HSD test resuled in significant difference between case1 and case2 groups (p = 0.038). Mice were monitored daily by personnel experienced in recognizing signs of morbidity (illness, injury, or abnormal behavior) also in weekends and holidays. Lab Anim 42, 217224 (2013). The caliper mean-percent-error is 10.6%, nearly twice that of the scanners mean-percent-error of 4.6%. Fj^P;7? Folia Biol. The problems with tumor volume measurements by calipers 3.1 Tumor growth: Height is not equal to the width of the tumor . Conceived and designed the experiments: ZS JS. The most recent ESMO (European Society for Medical Oncology) consensus guidelines suggest that treatment discontinuation or maintenance are feasible options after 46 months of full-dose first-line therapy to treat colon and rectal adenocarcinoma [37]. 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. The other way to measure tumor volume is the usage of small animal MRI, a non-invasive in vivo technology giving the possibility of a more precise volume measurement [29]. Internet J. Surg. The dose depends mainly on weight. Komrek, V. Gross anatomy. Applying the two-dimensional mathematical model (described in Eq 5) the goal is to find the f constant which belongs to the C38 colon adenocarcinoma and the treatment type. In the case2 group, one mouse died on the 8th day, before the therapy (bevacizumab administration) started. Our results for the case group (daily, quasi-continuous small amount administration) and the control group (one big dose according to the protocol) are 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 this way, the targeted therapy can be used in a much more individualized form. A smaller diameter mask will help to position the tumor in the center of the scan, but it will not alter this effect. Yes First is to find an appropriate mathematical model for tumor volume evaluation from caliper-measured data. However, if the treatment is lengthy, the problem of side-effects also has to be considered. recurrent glioblastoma (rGBM) [36]) as a first or second line treatment, usually in combination with chemo- or immunotherapy. Treat. In addition, the root mean square, R2 values under the linear regression equations in Figure 15. symbolize the precision of the data. % The widely accepted method for calculating the volume of subcutaneous tumors is to measure the length and width of the tumor with a caliper, and, while assuming that the tumor is the shape of an ellipsoid and that the height of the tumor is equal to its width, use the formula for the volume of an ellipsoid. 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. (2), Whilst studies have shown that tumors can be better estimated with ellipsoid shape than using Eq 1, calculating the volume of an ellipsoid requires the knowledge of the third parameter. 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. The volume above the red area is not occupied by the tumor, as seen in Figure 8. in The Laboratory Mouse (eds. ), the more correlated a given group (scanner or caliper estimations) is to the black line (r2 = 1), the more accurate the tumor volume measurements are. The post hoc test resulted in the following. Similarly, Zhang et al. When the tumors became palpable, measurements of tumor volume were performed (with calipers) once per week. Google Scholar. 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). In a study done in England, the researchers at the end of their experiment, dissected tumors and measured their weight with a precise balance, and used these weight values of the tumors to compare results of the caliper and the scanner measurements. 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. Mice in Phase I and Phase IV control group received no bevacizumab. @L Pk5*Gj@>Sk%SLL4w^c{j[hP(/h8HP_ @Ch( PASW Statistics 18 (SPSS Statistics, IBM, USA) and Matlab R2009b (MathWorks, USA) were used for statistical analysis. 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. 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. Predicting tumor volume in radical prostatectomy specimens from patients with prostate cancer. No, Is the Subject Area "Breast cancer" applicable to this article? Tumor growth can take place under such conditions; however delivery of chemotherapeutic drugs is obstructed. Mol. Our hypothesis is that the effectiveness of a lower dosage with a quasi-continuous therapy can be comparable with the protocol therapy. J. Clin. The effectiveness of bevacizumab administration was examined by statistical analysis. Orthotopic xenograft tumor models, which are more difficult to establish, are close to the clinical behavior of malignant tumors in terms of location of the primary tumor site, the tumor progression, the tumor micro-environment, cell migration and metastasis [1 . Lab Animal Even though all the three diameters can be measured, the estimation of the volume has quite a large error. 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. Preclinical evaluation of LU 79553: a novel bis-naphthalimide with potent antitumor activity. The TM900 also stores an optical image in the measurement data base. 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. HHS Vulnerability Disclosure, Help 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 . 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. How the device is positioned over the tumor will not influence the result. Weber, W.A. This results in the inclusion of sections of the parallelepipeds volume that does not exist in the actual tumor. 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. In order to assess interobserver variation, each . Tumor volume measurement was performed by digital caliper and small animal MRI. 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. Mammary tumors seemed to take on an oblate spheroid geometry. 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. (6) (7). 26 December 2022, Bulletin of Mathematical Biology A tumor image, its 3D shape and all geometrical data are automatically calculated and stored. Faustino-Rocha, A., Oliveira, P., Pinho-Oliveira, J. et al. The results presented here were provided by them. 41, 16901694 (1981). The tumor burden is evaluated by volume calculation using caliper measurement. Department of Control Engineering and Information Technology, Budapest University of Technology and Economics, Budapest, Hungary, Affiliation We found out that due to this unmeasured part, the dissected tumor volumes were about 6% larger than the scan results. 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. 120, 386391 (2003). Make sure you get accurate measurements for length, width and height. 2022 Dec 5;10:998013. doi: 10.3389/fchem.2022.998013. Tumor was located in the last third of the back of the mouse in every case. For more information about PLOS Subject Areas, click calipers and Water Displacement methods Tumor mimicking objects Vol. The first measurement in Phase I occurred on the 5th day; in Phase III/3, it occurred on the 4th day. Ergonomics 37, 217229 (1994). 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. Mice were killed after 38 days. Sport. We have found that there is significant difference between the means of the samples (p = 0.002, using 0.05 level of significance). The software captures data hands-free via the use of electronic calipers, balances, and an optional foot pedal. PubMedGoogle Scholar. Average of tumor volumes for every measurement days of the experiment can be seen in Fig 8. Russo, J. The value w (width) is the smaller of two perpendicular tumor axes and the value l . Tumor Volume Formula for Caliper Measurements, 3. 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. https://doi.org/10.1371/journal.pone.0142190.g004. Breast Cancer Res. In Figure 15., the linear regression equation for the best-fit line for VPleth values on the y-axis along with VC and VS values would be approximately y x, or y 1.00x + 0.00. 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. As it was discussed above, the handling of animals by calipers measurements also suffers such deformations of the animal body during the measurements. 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. Studies using other methods of imaging have exposed the problems that calipers pose. Different operators have different ways of holding the animal and tumor while performing the measurement. In Phase II, the toxicology investigation of the applied angiogenic inhibitor (bevacizumab) was performed; there was no tumor implantation into mice in this phase. The C38 colorectal carcinoma line was maintained by serial subcutaneous transplantations in C57Bl/6 mice. 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. This formula was the starting point of our examination to find an appropriate mathematical model. Edi3 in ER-HER2+ breast cancer effective blood supply due to the width of the data of. 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