This article is a commentary on:
Considering Intrauterine Location in a Model of Fetal Growth Restriction After Maternal Titanium Dioxide Nanoparticle Inhalation
John M. DeSesso
L. David Wise- 1Exponent Inc, Alexandria, VA, United States
- 2Independent Teratologist, Philadelphia, PA, United States
A Commentary on
Considering intrauterine location in a model of fetal growth restriction after maternal titanium dioxide nanoparticle inhalation
by DʼErrico JN, Fournier SB and Stapleton PA (2021). Front. Toxicol. 3:643804. doi: 10.3389/ftox.2021.643804
Introduction
We have read with interest the paper by D’Errico et al. (2021) and question the authors’ suggestion that fetal weights should be normalized based on their location within the uterus. Fetal weight effects are considered the most sensitive endpoint of embryo–fetal developmental toxicity (EFDT) studies (USEPA, 1991). Thus, it is important for a standard to be set for the calculation, presentation, and analysis of this endpoint.
To this end, it is well-established that the litter, rather than the fetus, is the experimental unit for analysis in EFDT studies (Jensh et al., 1970; Haseman and Hogan, 1975; USEPA, 1991; USEPA, 1998; OECD, 2018). Mean fetal weights are routinely calculated on a per litter basis wherein the mean fetal weight is calculated for each litter in a group, and then, these values are averaged to generate the overall group litter mean. Because male rat fetuses are generally heavier than females, mean fetal weights for each sex, calculated in the same general manner, are also typically reported. This method is considered the traditional dogma of fetal weight analyses.
An endpoint termed “normalized” or “adjusted” fetal weight is not described in regulatory guidance documents (e.g., USEPA, 1998; OECD, 2018). An inverse relationship between fetal body weight and litter size in polytocous species (although not absolute) has been recognized for many years (Romero et al., 1992) and should serve as the basis for developing methods that adjust fetal weights. The potential impact of uterine position on the distribution of nutrients and toxicants (and ultimately fetal health) has also been the subject of numerous investigations over the past 50 years (e.g., Barr et al., 1969; Barr et al., 1970; Ward et al., 1977; Norman and Bruce, 1979a; Norman and Bruce, 1979b; Padmanabhan and Singh, 1981). However, studies investigating the role of uterine position have produced conflicting results with regard to whether fetal weight is dependent or independent of horn size, horn side, and intrauterine position (Chahoud and Paumgartten, 2005).
Comments on the normalized fetal weight formula
D’Errico et al. (2021) used a novel method to adjust fetal weights to account for the uterine horn side and the number of fetuses per horn, according to the following formula:
Several issues arise when considering this approach. First, they reported normalized fetal weights in g; however, inspection of the aforementioned formula discloses that the calculated value is unitless. Interestingly, historical control data for GD 20 “traditional” fetal weights average > 3.5 g (Charles River, 2023), while their unadjusted fetal weight (2.5 g) is much smaller and the normalized weights (0.05 g) are unrealistically diminutive. Second, neither the use of terminal maternal body weight in the equation to adjust the fetal weight is justified nor is any reference cited to support this approach. The authors suggest that “future studies should utilize maternal weight gain throughout pregnancy;” however, while these data are typically collected, the additional data will not improve their approach. Regardless, maternal body weight gain is driven largely by the total weights of the fetuses and, therefore, is not a basis to adjust fetal weights. Third, the sex of the fetuses was admittedly not taken into consideration, although this is an important factor in determining the fetal weight. Fourth, the mean adjusted fetal weight reports at four horn positions per litter were calculated from only four fetuses in some groups, which is too low to reach valid conclusions. Lastly, data from 22% of all right horns and 29% of all left horns were excluded because of a rule that excluded uterine horns with fewer than five fetuses (i.e., due to complications within the formula). The exclusion of a quarter of the data is remarkable and calls into question the validity of both this approach for analyzing fetal weights and the conclusions.
The authors state “However, upon further evaluation using our revised approach, we observed that under control conditions, feti [sic] implanted in the left uterine horn tended to be smaller in body weight at term compared with feti [sic] implanted in the right uterine horn.” We note that their statement is based on a low number of control litters (20), of which five right uterine horns and eight left uterine horns were excluded due to very few fetuses, and that their conclusion is contradictory to that reported by Barr et al. (1970).
Conclusion
The approach taken by D’Errico et al. is indeed a challenge to accept as an alternative to the common dogma of litter data analysis. Given the numerous points raised above, we do not agree that this method in its present form allows for more refined conclusions to be drawn from fetal weight data. The substantial amount of the data that must be excluded to draw those conclusions from guideline-compliant EFDT studies would likely adversely impact human hazard identification.
Author contributions
JMD: writing–original draft and writing–review and editing. LW: writing–original draft and writing–review and editing.
Funding
The author(s) declare that no financial support was received for the research, authorship, and/or publication of this article.
Conflict of interest
Author JMD was employed by company Exponent Inc.
The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Publisher’s note
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References
Barr, M., Jensh, R. P., and Brent, R. L. (1969). Fetal weight and intrauterine position in rats. Teratology 2, 241–246. doi:10.1002/tera.1420020308
Barr, M., Jensh, R. P., and Brent, R. L. (1970). Prenatal growth in the albino rat: effects of number, intrauterine position and resorptions. Am. J. Anat. 128, 413–428. doi:10.1002/aja.1001280403
Chahoud, I., Ligensa, A., Dietzel, L., and Faqi, A. S. (1999). Correlation between maternal toxicity and embryo/fetal effects. Reprod. Toxicol. 13 (5), 375–381. doi:10.1016/s0890-6238(99)00035-0
Chahoud, I., and Paumgartten, F. J. R. (2005). Relationships between fetal body weight of Wistar rats at term and the extent of skeletal ossification. Braz J. Med. Biol. Res. 38, 565–575. doi:10.1590/S0100-879X2005000400010
Charles River (2023). Historical control data. https://www.criver.com/products-services/safety-assessment/toxicology-services/developmental-and-reproductive-toxicology-dart/historical-control-data?region=3601.
D’Errico, J. N., Fournier, S. B., and Stapleton, P. A. (2021). Considering intrauterine location in a model of fetal growth restriction after maternal titanium dioxide nanoparticle inhalation. Front. Toxicol. 3, 643804. doi:10.3389/ftox.2021.643804
Haseman, J. K., Bailer, A. J., Kodell, R. L., Morris, R., and Portier, K. (2001). Statistical issues in the analysis of low-dose endocrine disruptor data. Toxicol. Sci. 61, 201–210. doi:10.1093/toxsci/61.2.201
Haseman, J. K., and Hogan, M. D. (1975). Selection of the experimental unit in teratology studies. Teratology 12, 165–171. doi:10.1002/tera.1420120209
Jensh, R. P., Brent, R. L., and Barr, M. (1970). The litter effect as a variable in teratologic studies of the albino rat. Am. J. Anat. 128, 185–191. doi:10.1002/aja.1001280205
Norman, N. A., and Bruce, N. W. (1979a). Fetal and placental weight relationships in the rat at Days 13 and 17 of gestation. Reproduction 57 (2), 345–348. doi:10.1530/jrf.0.0570345
Norman, N. A., and Bruce, N. W. (1979b). Fetal and placental weight relationships in the albino rat near term. Teratology 19, 245–250. doi:10.1002/tera.1420190215
Organisation for Economic Cooperation and Development (OECD) (2018). in OECD guidelines for the testing of chemicals, section (Paris: OECD Publishing), 4. doi:10.1787/9789264070820-enTest No. 414: prenatal developmental toxicity study
Padmanabhan, P., and Singh, S. (1981). Effect of intrauterine position on foetal weight in CF rats. Indian J. Med. Res. 73, 134–139.
Romero, A., Villamayor, F., Grau, M. T., Sacristan, A., and Ortiz, J. A. (1992). Relationship between fetal weight and litter size in rats: application to reproductive toxicology studies. Reprod. Toxicol. 6, 453–456. doi:10.1016/0890-6238(92)90009-I
U. S. Environmental Protection Agency (USEPA) (1991). Guidelines for developmental toxicity risk assessment. Fed. Regist. 56 (234), 63798–63826.
U. S. Environmental Protection Agency (USEPA) (1998). Health effects test guidelines OPPTS 870.3700 prenatal developmental toxicity study. Washington, DC: U. S. Government Printing Office. EPA 712-C-98-207.
Keywords: rat, fetal weight, normalized, adjusted, EFDT studies
Citation: DeSesso JM and Wise LD (2023) Commentary: Considering intrauterine location in a model of fetal growth restriction after maternal titanium dioxide nanoparticle inhalation. Front. Toxicol. 5:1293873. doi: 10.3389/ftox.2023.1293873
Received: 13 September 2023; Accepted: 13 November 2023;
Published: 04 December 2023.
Edited by:
Rosaria Meccariello, University of Naples Parthenope, ItalyReviewed by:
Hideko Sone, Yokohama College of Pharmacy, JapanCopyright © 2023 DeSesso and Wise. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: John M. DeSesso, SmRlc2Vzc29AZXhwb25lbnQuY29t
†These authors have contributed equally to this work and share first authorship