For more information about PLOS Subject Areas, click The results presented here were provided by them. 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. Fornage, B.D., Toubas, O. 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). 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. Significance of rat mammary tumors for human risk assessment. Tumor morphology was investigated using standard Haematoxylin Eosin staining on the samples stored in formalin. Even though all the three diameters can be measured, the estimation of the volume has quite a large error. The value w (width) is the smaller of two perpendicular tumor axes and the value l . Harris, R.E., Alshafie, G.A., Abou-Issa, H. & Seibert, K. Chemopreventive of breast cancer in rats by celecoxib, a ciclooxygenase 2 inhibitor. Figure 8. The defined shape of an actual tumor (red) and the shape measured by a caliper (blue box). Fj^P;7? In vivo measurements of tumor metabolism and growth after administration of enzastaurin using small animal FDG positron emission tomography. One can find datasets in the last column of Table 2. Figure 17. N-nitrosomethylurea as mammary gland carcinogen in rats. 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. Mice in Phase I and Phase IV control group received no bevacizumab. Yes . Levenes test showed that the sample variances are equal (p = 0.108). obtained using vernier calipers (c.c) Vol. In the case of Phase I, no similar problem occurs; the two-dimensional mathematical model can handle great values as well (e.g. Funding: This work was supported by Hungarian Scientific Research Fund (OTKA), http://www.otka.hu/en, No: K-112993 (Jnos) and Bolyai Research Scholarship of the Hungarian Academy of Sciences, http://mta.hu/cikkek/?node_id=26057, No: 425_12. After fixing in methanol, rat anti-mouse CD31 antibody (1:50, 550274 BD PHARMINGEN) was applied as primer antibody, and FITC conjugated anti-rat CD31 antibody (1:100, Jackson Immunoresearch, 712-095-150) was applied on the slides as secondary antibody. Bevacizumab (Avastin) [1] is an exogenous inhibitor, which inhibits the biological activity of human VEGF [2]. Lee, J., Koh, D. & Ong, C.N. Pursuant to one-sample Kolmogorov-Smirnov test, each sample has normal distribution (ppIV_control = 0.981, ppIV_case1 = 0.998, ppIV_case2 = 0.854). Modern Pathol. 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). Open Access Materials and methods: The murine CaNT subcutaneous xenograft model was used. a) Haematoxylin Eosin (H&E) staining was applied to investigate tumor morphology. Figure 3. Further geometrical or visual data are not stored simultaneously. No, Is the Subject Area "Breast cancer" applicable to this article? Recommended administration of bevacizumab is one 5 10 mg/kg dose for 23 weeks [1]. Komrek, V. Gross anatomy. 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. n5). BMC Medical . An average penny has a diameter of 19 mm and a height of 1.27 mm, yielding a volume of 360.1 mm3. 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. 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. In Phase I, tumor growth was investigated without antiangiogenic therapy for 24 days (mice were sacrificed when the tumor reached a lethal size). You are using a browser version with limited support for CSS. & Robinson, S.P. Both methods cannot avoid variability introduced by the manipulation of the animal. 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 . J. Natl. by water displace ment (c.c) Vol. 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 19, 6170 (1989). & Wieder, H. Monitoring chemotherapy and radiotherapy of solid tumors. In Phase III/3, case group members received one-tenth dose of control dose intraperitoneally spread over 18 days. In this way, the targeted therapy can be used in a much more individualized form. 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. Wrote the paper: JS ZS PK LK. Starting with f = 1, we have investigated the goodness of the fitting, using an iterative method. 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. First way is the caliper measurement; in that case tumor diameters (width, length) are measured with digital caliper. This process not only deforms the tumor mass but also creates undesirable pain on the animal. Am. 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. 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). 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. Tumor volume in subcutaneous mouse xenografts measured by microCT is more accurate and reproducible than determined by 18 F-FDG-microPET or external caliper. Experimentally induced mammary tumors in rats. Figure 6. This means that the daily treatment with one-twelfth total dose resulted in significantly smaller tumors than the protocol-based treatment. In the case2 group, one mouse died on the 8th day, before the therapy (bevacizumab administration) started. These images were applied to calculate vascularization area by using ImageJ (NIH, USA) [33] software (Fig 4). Mice were monitored daily by personnel experienced in recognizing signs of morbidity (illness, injury, or abnormal behavior) also in weekends and holidays. These images also provide extra morphological information about the tumor growth and the effect of the compounds on tumor behavior. The volume calculation algorithm relies on the surface interpolated based on the surrounding healthy tissue. In addition, the mice were weighed . 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. In this study, the authors measured dimensions of rat mammary tumors using a caliper and using real-time . The significant difference between quasi-continuous therapy (Phase III/3 case group) and tumor growth without treatment (Phase I) was proved with statistical analysis as well. The mathematical relationship between MRI-measured tumor volume and mass was investigated to estimate accurate tumor volume using caliper-measured data. . This results in the inclusion of sections of the parallelepipeds volume that does not exist in the actual tumor. Autopsy revealed only minimal local tumor; however death was not caused by tumor. 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. Using a caliper, growth of the tumor in height is not measured as only width and length changes will affect the calculated volume. Lab Animal CAS Linear regression analysis to identify an equation that can be used to calculate tumor depth from tumor length as measured by caliper. Radiology 233, 830849 (2004). Because the industrys caliper-based formula allows for these inclusions in nonexistent volumes more often than not, it yields inaccurate measurements, which can confuse. Unable to load your collection due to an error, Unable to load your delegates due to an error. Typical side-effects and adverse events are arterial and venous thromboembolic events, bleeding, hypertension, febrile neutropenia, infections, proteinuria, mucositis, and hand-foot syndrome. Notice that the caliper-based measurements have a larger standard deviation. Tumor volume measurement cannot happen right after tumor implantation. Conceived and designed the experiments: ZS JS. We have found that there is significant difference between the means of the samples (p = 0.002, using 0.05 level of significance). 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. 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. In Phase I, we have investigated tumor growth without therapy with two types of mouse tumor. 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]. 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. <> 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. Analysis of Variance (ANOVA) test was used to compare more than two samples, while pairwise comparison was done by Tukeys honest significant difference (HSD) test. Figure 1. where V m is the measurement threshold that represents the smallest measurable tumor volume using a caliper. In the meantime, to ensure continued support, we are displaying the site without styles 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. & trukelj, B. N-methylnitrosourea induced breast cancer in rat, the histopathology of the resulting tumours and its drawbacks as a model. 39, 720 (1996). Parkin, D.M., Bray, F., Ferlay, J. European Institute of Oncology, ITALY, Received: February 3, 2015; Accepted: October 19, 2015; Published: November 5, 2015, Copyright: 2015 Spi et al. Effects of tamoxifen and melatonin on mammary gland cancer induced by N-methyl-N-nitrosourea and by 7,12-dimethylbenz(a)anthracene, respectively, in female Sprague-Dawley rats. Timing wise both methods perform the same. [2]no matter how accurately the length and width of the tumor are measured. Tumors should also be placed so that the tumor peak is near the center of the mask and is not highly inclined. Enter your measurements into the Tumor Volume Calculator to determine the volume of the tumor. (5) This process takes 45 days. Similarly, Zhang et al. 55, 74108 (2005). Mice were killed after 38 days. To compare the effectiveness of the different investigated treatment methods, statistical analysis was used. <> stream Sci Rep. 2022 Dec 26;12(1):22336. doi: 10.1038/s41598-022-25258-3. Yes J Clin Oncol. and calculates an interpolated surface under the tumor boundary based on the healthy surface around the tumor (navy blue line in Figure 12.). 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, case2 group members received 1.11 g bevacizumab (with 45 l 0.9% NaCl solution) every day form the 8th day for 15 days. Modulation of mammary tumor vascularization by mast cells: Ultrasonographic and histopathological approaches. Mice in Phase III/3 control and case groups, and mice in Phase IV case1 and case2 groups received bevacizumab. Furthermore, caliper-based measurements cause undue stress on the animal and, by extension, the tumor, which can further distort measurements. https://doi.org/10.1371/journal.pone.0142190.t003. in The Laboratory Mouse (eds. Determining tumor volume is a useful and quantitative way to monitor tumor progression. A tumor expands in width rather than height as it grows. 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%. Measure the three dimensions of peanut M&M's, serving as our similar-sized tumor surrogates, using vernier calipers. Mammary tumors similar to those observed in women can be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea. et al. Davis, P.L. 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. This can happen if the mouse is pressed into the mask or if the body is stretched when held. Epub 2013 Jul 11. Cancer 83, 621628 (1998). produced by the TumorImager-2TM so that the animal is aligned under the mask each time similarly to the previous scan. Breast Cancer Res. PMID: 11208850. Objectives: Volume measurement of subcutaneous xenograft tumors in nude mice models is an important metric to assess tumor growth or response to therapy. MeSH by US (c.c) Vol. How the device is positioned over the tumor will not influence the result. 105, 2023 (1983). Carlsson, G., Gullberg, B. The TumorImagerTM line of scanners by Biopticon Corporation fixes these problems by keeping the animals relaxed during measuring and yielding measurements closer to actual values than caliper-based measurements. Tumor size was measured every 3 days using a digital caliper and computed according to the ellipsoidal calculation formula: V . Physical activity cancer preventionmechanisms. Sport. One can find datasets in Table 3. 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). 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 volume above the red area is not occupied by the tumor, as seen in Figure 8. Get the most important science stories of the day, free in your inbox. In this phase, three groups were created. Traditionally, caliper measurements are used to calculate tumor volume. PubMedGoogle Scholar. In the case of Phase I and Phase IV, there were no MRI measurements. Usually, 3.0 x 10 6 cells are needed per injection. 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