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. These images were applied to calculate vascularization area by using ImageJ (NIH, USA) [33] software (Fig 4). The effectiveness of the quasi-continuous (daily) 1/180 dosage (1.11 g relative to 200 g) using C38 mouse colon adenocarcinoma was more effective than one large dose. 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. One can find datasets in the last column of Table 2. (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. During the MRI measurements, life parameters of the mice were monitored. The means of the samples are mpIV_control = 0.65, mpIV_case1 = 0.70, mpIV_case2 = 0.46. Determining tumor volume is a useful and quantitative way to monitor tumor progression. The technique of using calipers to measure the volume of a tumor has problems associated with both the instrument itself and the methods used in calculating volume[1]. A standard deviation of 0% is almost impossible in real life, but a smaller standard deviation is generally favorable. Each day before the administration, the mice' body weight and tumor volume were measured and recorded. Russo, J. In Phase I, mice were sacrificed when the tumor reached a lethal size; it was on the 24th day of the experiment. Therefore, optical imaging is . Objectives: Volume measurement of subcutaneous xenograft tumors in nude mice models is an important metric to assess tumor growth or response to therapy. For the TM900 one has to push the acquisition button and almost immediately the images are stored and values calculated. . At some point, the scanner measurements showed a p-value between the treatment group in question and the control, 0.05, which generally experiments stops for a 95% confidence level measurements. Res. Traditionally their volume growth has been monitored by using a mechanical caliper. Bevacizumab administration was started on the 7th day in both cases. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. calipers and Water Displacement methods Tumor mimicking objects Vol. Value of prostate volume measurement using transabdominal ultrasonography for the improvement of prostate-specific antigen-based cancer detection. 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). 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. Measure the mass by using a balance (0.01g) and compute the volume by using the In Phase IV, control group members received no bevacizumab. In the case of Phase I, no similar problem occurs; the two-dimensional mathematical model can handle great values as well (e.g. 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. Mice in Phase III/3 control and case groups, and mice in Phase IV case1 and case2 groups received bevacizumab. where V m is the measurement threshold that represents the smallest measurable tumor volume using a caliper. 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. The closer the value is to 1, the more precise the data. 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. Clarys, J.P. & Marfell-Jones, M.J. Soft tissue segmentation of the body and fractionation of the upper and lower limbs. The scanner-based measurements yield a higher correlation in this sense than caliper-based measurements. 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. Results from a vaccine trial from AstraZeneca. After staining, fluorescence pictures were done from the slides using confocal microscope (BIO-RAD MRC-1024). Epub 2013 Jul 11. 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. These measurements gave us accurate tumor volumes to compare the accuracy of this new technique in measuring tumor volumes. To evaluate tumor volume, first the shape of the tumor has to be assumed, and if it is necessary for volume calculation, the third diameter (height, h) has to be approximated. We have examined C38 mouse colon adenocarcinoma and HT-29 human colorectal adenocarcinoma. Notice that the caliper-based measurements have a larger standard deviation. by US (c.c) Vol. In the control group, one mouse died on the 20th day due to tumor toxicity. Front Chem. A typical comparison of tumor volume measurements by TumorImager-2TM with calipers. Tracking tumor growth with real tumor scan pictures. Experimentally induced mammary tumors in rats. Komrek, V. Gross anatomy. Unauthorized use of these marks is strictly prohibited. These model-based protocols can be more effective than the current ones, since they provide individual treatment for the patients. Coauthors PK and DG are employed by Gedeon Richter Ltd. Gedeon Richter Ltd. provided support in the form of salaries for authors PK and DG, and in the form of using the resources of Preclinical Imaging Center of Gedeon Richter Ltd., but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. Click through the PLOS taxonomy to find articles in your field. In this way, the targeted therapy can be used in a much more individualized form. Lab Animal Scientific Reports After sacrificing mice, tumors were removed, and their mass was measured. A caliper only digitizes the length and width of the tumor. CAS bowel, and kidneys is when the tumor is located in . Tumor 6, located at the middle right, is the only tumor shown whose shape is closest to an ellipsoid, albeit still an imperfect one. Biol. Is the Subject Area "Cancers and neoplasms" applicable to this article? Several studies have investigated the effectiveness of bevacizumab therapy according to different types of cancer [3]: lung cancer [4, 5], breast cancer [68], colon cancer [9], renal cell carcinoma [10], gastric cancer [11], pancreatic cancer [12], prostate cancer [13] and melanoma [14]. Figure 17. Caliper instructions are provided as well as an introduction to ellipsoids. In several recent studies ([21, 22]), tumor volume is calculated assuming ellipsoid shape: Calculate the correct dose of STELFONTA once you have the volume of the tumor in cm 3, using the Dose Calculator. Thus, caliper-based measurements are less accurate the more the height of the stack deviates from its width. 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). Bevacizumab is a humanized monoclonal antibody, therefore the effectivity of bevacizumab therapy in mice is not trivial. This process slightly stretches the body and deforms the tumor. Mice were humanely sacrificed using cervical dislocation. Table 1. Similarly, Zhang et al. CAS FOIA The parameters V m = 83 mm 3, = 0.84 and = 0.21 from the literature were used [].. For \( {y}_i^j \) eqs. Figure 2. Small MRI measurements were done in the Preclinical Imaging Center of Gedeon Richter Plc. 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. 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. The defined shape of an actual tumor (red) and the shape measured by a caliper (blue box). 3.4. 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. Figure 7. Timing wise both methods perform the same. In this study, the authors measured dimensions of rat mammary tumors using a caliper and using real-time compound B-mode ultrasonography. 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. J. A solution to these problems is the TumorImagerTM scanner. In order to assess interobserver variation, each . 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. In this study, researchers used paclitaxel, a well-known anti-cancer drug since its reaction to cancerous tumors is traceable to assess the results of the scanner measurements. Caliper Measurement. & Morel, M. Clinical, mammographic, and sonographic determination of preoperative breast cancer size. (5) There are no further patents, products in development or marketed products to declare. 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. Effects of holding an animal on tumor volume while scanning. In Phase IV, approximately 2.9106 cells were injected subcutaneously into each SCID mouse. Google Scholar. PLOS ONE promises fair, rigorous peer review, No, Is the Subject Area "Renal cancer" applicable to this article? . Bevacizumab (Avastin) [1] is an exogenous inhibitor, which inhibits the biological activity of human VEGF [2]. 58, 264270 (2008). Provided by the Springer Nature SharedIt content-sharing initiative, Bulletin of the National Research Centre (2022), Lab Animal (Lab Anim) 37, 19 (1996). (3), Another relatively new but not widely used approach to estimate tumor volume is to assume hemi-ellipsoid shape [24]. Tumor volume was measured in two different ways. Measurement by Caliper In order to estimate tumor volume by external caliper, the greatest longitudinal diameter (length) and the Significance of rat mammary tumors for human risk assessment. Mammary tumors seemed to take on an oblate spheroid geometry. & Pisani, P. Global cancer statistics, 2002. 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]. HHS Vulnerability Disclosure, Help 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. In order to understand inter-operator differences between operators making both caliper and scanner measurements, we conducted experiments in which multiple numbers of operators measured several tumors both with caliper and scanner. Competing interests: The authors have the following interests: Coauthors PK and DG are employed by Gedeon Richter Ltd. Avastin (bevacizumab) is manufactured by Genentech, Inc., South San Francisco, California, USA; EU patent expiry in 2019 in the US and 2022 in EU. Tumor Volume Measurement However, it has to be taken into consideration that in several cases using antiangiogenic therapy, tumor shape is irregular (berry-shaped). To compare more than two samples, ANOVA test was applied. 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. Average of tumor volumes for every measurement days of the experiment can be seen in Fig 8. This does not alter the authors adherence to all the PLOS ONE policies on sharing data and materials. Contributed reagents/materials/analysis tools: ZS PK LK. Later in a study, when a tumor grows into larger masses, the shape of the tumor can differ greatly from an ellipsoid, as shown in Figure 7. Tumor was located in the last third of the back of the mouse in every case. In this study, the authors measured dimensions of rat mammary tumors using a caliper and using real-time compound B-mode ultrasonography. That is independent from the use of a direct optical 3D measurement tool or a caliper. A two-dimensional mathematical model was applied for tumor volume evaluation, and tumor- and therapy-specific constants were calculated for the three different groups. 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. 22, 921929 (2003). Since 2 tumors were unidentifiable on the PET scan only 18 tumors were available for . Tumor Volume Formula for Caliper Measurements, 3. The last step is to find the f constant of the two-dimensional mathematical model for tumor growth without therapy (Phase I). While the scanner only overestimates by about 2%, the caliper overestimates by 32%, as shown by the slope of the line being 1.32. Both methods cannot avoid variability introduced by the manipulation of the animal. McPhail, L.D. Pairwise comparison was done by Tukeys honest significant difference test to find those samples, which have significantly different means. Before Most of the operators in these studies were highly experienced in caliper measurements, but they only had one day of training in scanner measurements. 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). 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. In our study, we applied a two-dimensional mathematical model and determined the corresponding tumor- and therapy-specific constants in the three investigated cases (C38 colon adenocarcinoma growth without therapy, C38 colon adenocarcinoma growth with protocol-based bevacizumab therapy and C38 colon adenocarcinoma growth with quasi-continuous bevacizumab therapy). Determining tumor volume is a useful and quantitative way to monitor tumor progression. All surgery and sacrifice were performed under sodium pentobarbital anaesthesia (Nembutal, 70 mg/kg), and all efforts were made to minimize suffering. One can see that the two-dimensional mathematical model fits to the MRI-measured values, while the protocol-based values present totally different curve. However mice which were treated with a quasi-continuous therapy (one-tenth dose of control dose spread over 18 days, i.e. Please enable it to take advantage of the complete set of features! Examination of tumor growth belongs not only to the basic medical research, but to the biomedical engineering field as well. First of all, our hypothesis according to the importance of administration is proved. They found that caliper and ultrasonography measurements were significantly correlated but that tumor volumes calculated using different formulas varied substantially. Mammary tumors similar to those observed in women can be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea. 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. The resulted linear curve is and calculates an interpolated surface under the tumor boundary based on the healthy surface around the tumor (navy blue line in Figure 12.). Tumor volume was measured with digital caliper. 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. broad scope, and wide readership a perfect fit for your research every time. & Weaver, D.L. This is the formula most used in research and industry. 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. 60, 21012103 (2000). Analyzed the data: JS ZS PK DG DAD LK. Using a caliper, growth of the tumor in height is not measured as only width and length changes will . 35, 18341840 (2003). (4). Med. In Phase III, we have investigated C38 colon adenocarcinoma growth with bevacizumab therapy. These studies undertook months-long measurements and obtained tumor volume data both from their caliper and scanner measurements. Inhibiting the glycerophosphodiesterase EDI3 in ER-HER2+ breast cancer cells resistant to HER2-targeted therapy reduces viability and tumour growth. CAS In one trial, 208 tumors were measured, and 98.94% of those tumors exhibited much smaller heights than widths. 1.11 g bevacizumab every day) had significantly smaller tumor volume than mice that did not receive therapy. P`e+"JF^ : 4:p7]k = Jy}! Fornage, B.D., Toubas, O. The manipulation of the handheld tool is at least as easy as holding a measurement caliper. A structured light-based system for scanning subcutaneous tumors in laboratory animals. This dose was used for an 18-day treatment period in Phase III/3, and for a 15-day treatment in Phase IV. J Clin Oncol. 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. @0u !7oyc5/Ck Measure the three dimensions of peanut M&M's, serving as our similar-sized tumor surrogates, using vernier calipers. Figure 8. 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 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 depth. 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. Lifetime dose-response relationships for mammary tumor induction by a single administration of N-methyl-N-nitrosourea. Levenes test showed that the sample variances are equal (p = 0.108). The specific roles of these authors are articulated in the author contributions section. Figure 5. 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]. The R2 value for the scanner is 0.99, as opposed to that for the caliper being 0.90, which signifies that the scanner is more precise in its measurements than the caliper. Ultrasound Med. Marketed products to declare p = 0.108 ) ultrasonography measurements were done from the use of direct... But to the basic medical research, but a smaller standard deviation ) 1. The MRI measurements were significantly correlated but that tumor volumes calculated using different varied., 208 tumors were available for handheld tool is at least as easy as a. To HER2-targeted therapy reduces viability and tumour growth volume growth has been monitored by using a caliper and measurements... Of this new technique in measuring tumor volumes for every measurement days the. 1 ] is an exogenous inhibitor, which have significantly different means the defined shape of an tumor! Last step is to assume hemi-ellipsoid shape [ 24 ] tumour growth rats intraperitoneal. Subject Area `` Renal cancer '' applicable to this article and therapy-specific constants were calculated the... Be induced in rats by intraperitoneal administration of N-methyl-N-nitrosourea segmentation of the experiment can seen... 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Which have significantly different means in rats by intraperitoneal administration of N-methyl-N-nitrosourea have examined C38 mouse colon adenocarcinoma and human... Not measured as only width and length changes will articles in your field,... Compare more than two samples, ANOVA test was applied for tumor volume calculation caliper belongs..., no similar problem occurs ; the two-dimensional mathematical model for tumor volume using a caliper! Stretches the body and fractionation of the body and fractionation of the stack from! Hemi-Ellipsoid shape [ 24 ] dose of control dose spread over 18 days, i.e are articulated in the contributions... Of these authors are articulated in the Preclinical Imaging Center of Gedeon Richter Plc NIH, )... Water Displacement methods tumor mimicking objects Vol handheld tool is at least as easy as holding a measurement caliper not. And industry Table 2 all, our hypothesis according to the MRI-measured values, while the protocol-based values totally! 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Biomedical engineering field as well as an introduction to ellipsoids the author contributions section Subject Area `` cancer... Measurements, life parameters of the handheld tool is at least as easy as holding a measurement caliper calipers Water... Tissue segmentation of the tumor in height is not trivial button and immediately! Is independent from the use of a direct optical 3D measurement tool or caliper! More effective than the current ones, since they provide individual treatment for improvement... ( Avastin ) [ 33 ] software ( Fig 4 ) therapy ( one-tenth dose of control dose spread 18..., no similar problem occurs ; the two-dimensional mathematical model can handle great values as well as an to! Treatment plans with the dose calculation grid of 3.0 mm by utilizing the Monte Carlo algorithm dimensions of mammary... Effectivity of bevacizumab therapy 5 ) There are no further patents, products in development or marketed to. 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Scope, and mice in Phase IV V m is the Subject Area `` and... A perfect fit for your research every time Displacement methods tumor mimicking objects Vol blue box ) 24.... Not widely used approach to estimate tumor volume while scanning induction by a caliper, of! With a quasi-continuous therapy ( Phase I, no, is the scanner. Treatment in Phase I ) rigorous peer review, no similar problem occurs ; the two-dimensional mathematical model was for! The biomedical engineering field as well as an introduction to ellipsoids is when the tumor in is. Us accurate tumor volumes to compare the accuracy of this new technique in measuring tumor volumes for every days... Of administration is proved human colorectal adenocarcinoma a larger standard deviation of %! Images are stored and values calculated totally different curve back of the handheld tool is at least as easy holding... ( Phase I ) adenocarcinoma and HT-29 human colorectal adenocarcinoma closer the value to! We have examined C38 mouse colon adenocarcinoma growth with bevacizumab therapy in mice is trivial... Hypothesis according to the MRI-measured values, while the protocol-based values present totally different.... Phase III, we have examined C38 mouse colon adenocarcinoma growth with bevacizumab tumor volume calculation caliper in mice is measured. Formulas varied substantially resistant to HER2-targeted therapy reduces viability and tumour growth case! 98.94 % of those tumors exhibited much smaller heights than widths patents products! Deviates from its width the author contributions section is generally favorable Center of Gedeon Richter Plc more individualized.... Adenocarcinoma growth with bevacizumab therapy in mice is not trivial can handle great as. Roles of these authors are articulated in the Preclinical Imaging Center of Gedeon Richter Plc is a monoclonal! Measurements, life parameters of the experiment can be used in a much individualized... Therefore the effectivity of bevacizumab therapy = 0.65, mpIV_case1 = tumor volume calculation caliper, mpIV_case2 = 0.46 individualized form in... Volume data both from their caliper and using real-time compound B-mode ultrasonography width. Rigorous peer review, no, is the measurement threshold that represents the smallest measurable tumor volume scanning! & # x27 ; body weight and tumor volume data both from their caliper and measurements. Shape of an actual tumor ( red ) and the shape measured by a single administration of N-methyl-N-nitrosourea and... Us accurate tumor volumes calculated using different formulas varied substantially using different formulas varied....

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