Formulation of mice diet with low cholecalciferol content
DOI:
https://doi.org/10.13181/mji.oa.257859Keywords:
diet, food safety, methods, mice, vitamin DAbstract
BACKGROUND Vitamin D deficiency has been linked to autoimmune diseases, cancer, and cardiovascular diseases. Although 1 study attempted to elucidate the ingredients required to make this diet, the process remained unclear. Hence, this study aimed to customize a low cholecalciferol diet with good tolerability in mice.
METHODS We customized a diet containing a normal cholecalciferol content
(1 IU/g diet) and another with a low cholecalciferol content (0.05 IU/g diet). Samples from both diets were sent to an independent laboratory to ensure that the levels of cholecalciferol, phosphorus, and calcium present in the custom diets matched our calculations. 5 mice were fed the customized normal cholecalciferol diet for 1 week to assess tolerability. Tolerability was assessed by measuring the amount of food consumed, weight gained, and the presence of any adverse events.
RESULTS Cholecalciferol, phosphorous, and calcium levels in both diets satisfactorily matched our calculations. The diet was well tolerated without any adverse events or mortalities. The mice consumed an adequate amount of food (mean: 5.34 [0.08] g diet/day, 95% confidence interval [CI]: 5.12–5.56; 19.38 kcal, fat: 0.43 g, protein: 0.14 g, carbohydrates: 3.16 g, and cholecalciferol: 0.007 mg) and gained a slight amount of weight by the end of the experiment (mean: 1.86 [0.46] g, 95% CI: 0.58–3.14).
CONCLUSIONS This study successfully created 2 custom diets with quantified cholecalciferol contents. This animal model may prove valuable for studies involving vitamin D.
Downloads
References
Athanassiou L, Kostoglou-Athanassiou I, Koutsilieris M, Shoenfeld Y. Vitamin D and autoimmune rheumatic diseases. Biomolecules. 2023;13(4):709. https://doi.org/10.3390/biom13040709
Mallya SM, Corrado KR, Saria EA, Yuan FF, Tran HQ, Saucier K, et al. Modeling vitamin D insufficiency and moderate deficiency in adult mice via dietary cholecalciferol restriction. Endocr Res. 2016;41(4):290−9. https://doi.org/10.3109/07435800.2016.1141937
Ma J, Wu D, Li C, Fan C, Chao N, Liu J, et al. Lower serum 25-hydroxyvitamin D level is associated with 3 types of autoimmune thyroid diseases. Medicine (Baltimore). 2015;94(39):e1639. https://doi.org/10.1097/MD.0000000000001639
Barragan M, Good M, Kolls JK. Regulation of dendritic cell function by vitamin D. Nutrients. 2015;7(9):8127−51. https://doi.org/10.3390/nu7095383
Young MR, Xiong Y. Influence of vitamin D on cancer risk and treatment: why the variability? Trends Cancer Res. 2018;13:43−53.
Haider F, Ghafoor H, Hassan OF, Farooqui K, Bel Khair AO, Shoaib F. Vitamin D and cardiovascular diseases: an update. Cureus. 2023;15(11):e49734. https://doi.org/10.7759/cureus.49734
Di Franco M, Barchetta I, Iannuccelli C, Gerardi MC, Frisenda S, Ceccarelli F, et al. Hypovitaminosis D in recent onset rheumatoid arthritis is predictive of reduced response to treatment and increased disease activity: a 12 month follow-up study. BMC Musculoskelet Disord. 2015;16:53. https://doi.org/10.1186/s12891-015-0505-6
Filoni A, Vestita M, Congedo M, Giudice G, Tafuri S, Bonamonte D. Association between psoriasis and vitamin D: duration of disease correlates with decreased vitamin D serum levels: an observational case-control study. Medicine (Baltimore). 2018;97(25):e11185. https://doi.org/10.1097/MD.0000000000011185
Ospina-Caicedo AI, Cardona-Rincón AD, Bello-Gualtero JM, Valle-Oñate R, Romero-Sánchez C, Chalem-Choueka P, et al. Lower levels of vitamin D associated with disease activity in colombian patients with systemic lupus erythematosus. Curr Rheumatol Rev. 2019;15(2):146−53. https://doi.org/10.2174/1573397114666181015161547
O'Brien KM, Sandler DP, Taylor JA, Weinberg CR. Serum vitamin D and risk of breast cancer within five years. Environ Health Perspect. 2017;125(7):077004. https://doi.org/10.1289/EHP943
Shaukat N, Jaleel F, Moosa FA, Qureshi NA. Association between vitamin D deficiency and breast cancer. Pak J Med Sci. 2017;33(3):645−9. https://doi.org/10.12669/pjms.333.11753
Lewis C, Xun P, He K. Vitamin D supplementation and quality of life following diagnosis in stage II colorectal cancer patients: a 24-month prospective study. Support Care Cancer. 2016;24(4):1655−61. https://doi.org/10.1007/s00520-015-2945-9
Hu B, Chen J, Shi Y, Hou L. Association between serum 25(OH)D and risk of all-cause mortality in adults with prior cardiovascular disease: a cohort study from NHANES 2007-2018. BMC Cardiovasc Disord. 2023;23(1):240. https://doi.org/10.1186/s12872-023-03257-0
Yang J, Dong Y, Naugler CT, de Koning L. Serum 25-hydroxyvitamin D, cardiovascular risk markers, and incident cardiovascular disease in a high risk community population. Clin Biochem. 2021;93:36−41. https://doi.org/10.1016/j.clinbiochem.2021.04.001
Zhou Y, Jiang M, Sun JY, Cheng C, Shen H, Sun W, et al. The association between vitamin D levels and the 10-year risk of atherosclerotic cardiovascular disease: a population-based study. J Cardiovasc Nurs. 2023;38(5):E178−86. https://doi.org/10.1097/JCN.0000000000000943
Holick MF. The vitamin D deficiency pandemic: approaches for diagnosis, treatment and prevention. Rev Endocr Metab Disord. 2017;18(2):153−65. https://doi.org/10.1007/s11154-017-9424-1
Kraemer AN, Schäfer AL, Sprenger DT, Sehnert B, Williams JP, Luo A, et al. Impact of dietary vitamin D on immunoregulation and disease pathology in lupus-prone NZB/W F1 mice. Front Immunol. 2022;13:933191. https://doi.org/10.3389/fimmu.2022.933191
Reynolds JA, Rosenberg AZ, Smith CK, Sergeant JC, Rice GI, Briggs TA, et al. Brief report: vitamin D deficiency is associated with endothelial dysfunction and increases type i interferon gene expression in a murine model of systemic lupus erythematosus. Arthritis Rheumatol. 2016;68(12):2929−35. https://doi.org/10.1002/art.39803
Yamamoto EA, Nguyen JK, Liu J, Keller E, Campbell N, Zhang CJ, et al. Low levels of vitamin D promote memory B cells in lupus. Nutrients. 2020;12(2):291. https://doi.org/10.3390/nu12020291
Weiskirchen S, Weiper K, Tolba RH, Weiskirchen R. All you can feed: some comments on production of mouse diets used in biomedical research with special emphasis on non-alcoholic fatty liver disease research. Nutrients. 2020;12(1):163. https://doi.org/10.3390/nu12010163
The Jackson Laboratory. Body weight information for BALBc (000651): JAX® mice strain BALB/cJ [Internet]. The Jackson Laboratory; 2025 [cited 2023 Oct 23]. Available from: https://www.jax.org/jax-mice-and-services/strain-data-sheet-pages/body-weight-chart-000651.
VK MA, VK Kalaichelvan, VV Venkatachalam. Acute and subacute toxicity assessment of ethyl acetate extracts from aerial parts of Clerodendrum thomsoniae Balf.f in rodents. Biointerface Res Appl Chem. 2021;11(6):13952−61. https://doi.org/10.33263/BRIAC116.1395213961
U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition. Guidance for industry and other stakeholders: redbook 2000: toxicological principles for the safety assessment of food ingredients. U.S. Food and Drug Admininstration, Center for Food Safety and Applied Nutrition; 2007.
Orabueze IC, Olufunmi HO, Ota DA, Asare G. Co-administration effects and toxicity profile of ethyl acetate fraction of Stemonocoleus micranthus harms and artesunate in murine malaria model. S Afr J Bot. 2022;146:715−22. https://doi.org/10.1016/j.sajb.2021.12.005
Dominguez LJ, Farruggia M, Veronese N, Barbagallo M. Vitamin D sources, metabolism, and deficiency: available compounds and guidelines for its treatment. Metabolites. 2021;11(4):255. https://doi.org/10.3390/metabo11040255
Fleet JC. Vitamin D-mediated regulation of intestinal calcium absorption. Nutrients. 2022;14(16):3351. https://doi.org/10.3390/nu14163351
Ozturk B, Argin S, Ozilgen M, McClements DJ. Nanoemulsion delivery systems for oil-soluble vitamins: influence of carrier oil type on lipid digestion and vitamin D3 bioaccessibility. Food Chem. 2015;187:499−506. https://doi.org/10.1016/j.foodchem.2015.04.065
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Sumariyono, Dian Handayani, Siti Setiati, Nadia Amalialjinan, Gaby Abellia, Risma Debby Anindyanti, Syifa Sarah Dienillah, Ernawati, Caroline Tanadi

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with Medical Journal of Indonesia agree to the following terms:
- Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
- Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.



