N 2 (199) 2026. P. 112–118

SAFETY ASSESSMENT OF MINOXIDIL LOTIONS AND HEMODIALYSIS SOLUTIONS: ALTERNATIVE TOXICITY TESTING METHODS (HET-CAM METHOD)

N. Ye. Chemodurova, V. A. Turkina, A. M. Filipska, N. I. Hudz

State Nonprofit Company “Danylo Halytsky Lviv National Medical University”, Lviv, Ukraine
University of Opole, Opole, Poland

DOI 10.32782/2226-2008-2026-2-16

Introduction. The HET-CAM test was developed for the assessment of the irritation potential of cosmetic, chemical and pharmaceutical products. A good correlation between the results of the HET-CAM test and the in vivo test was found in numerous experiments.

The aim of this work was to evaluate the irritant effect of 5% and 10% minoxidil lotions and acetic and citric acid hemodialysis concentrates on chicken embryo chorioallantoic membrane using a digital microcamera to objectify the research results.

Materials and research methods. Incubated eggs were used. Vessel lysis, hemorrhage, and coagulation were monitored at the chorioallantoic membrane. According to the degree and time of vascular damage the irritation caused by the substance under study was evaluated. The resulting irritation index is used as a criterion for the classification of the irritant activity of the tested substance. Data were described using the median (Me) and interquartile range.

Results and their discussion. After exposure to a lotion containing 5% minoxidil, vessel lysis began at 120 seconds, spot hemorrhages – at 30 seconds in all observations; coagulation – at 120 seconds in one observation. Exposure to a lotion containing 10% minoxidil caused vascular bleeding at 30 seconds in one observation, at 120 seconds іn two observations, coagulation at 30 seconds and the lysis – at 120 seconds in all observations. Hemodialysis concentrates with acetic acid had a moderate irritating effect, and citric acid had a mild irritating effect.

Conclusions. The implementation of modern digital technologies of test visualization in the HET-CAM test allows for highly detailed photo-fixation and accurate interpretation of the results. It proves its effectiveness, expediency and informative value, which contributes to the improvement of alternative research methods.

Keywords: irritant effect, HET-CAM test, minoxidil, hemodialysis concentrates.

REFERENCES

  1. Anand Babu K, Ram Narayanan R. Determination of ocular irritancy potential of ophthalmic products using HET-CAM method. Research Journal of Pharmacy and Technology. 2021;14(6):3063–3066. https://doi.org/10.52711/0974-360X.2021.00535.
  2. Smail SS. Ex vivo irritation evaluation of a novel brimonidine nanoemulsion using the Hen’s egg test on chorioallantoic membrane (HET-CAM). Cureus. 2024;16(8):e68280. https://doi.org/10.7759/cureus.68280.
  3. Nicolás R, Lenze MM, Izaguirre M, Perez Damonte S, Aguilar A, Wikinski S, et al. Comparison between HET-CAM protocols and a product use clinical study for eye irritation evaluation of personal care products including cosmetics according to their surfactant composition. Food and Chemical Toxicology. 2021;153:112229. https://doi.org/10.1016/j.fct.2021.112229.
  4. Schrage A, Gamer AO, van Ravenzwaay B, Landsiedel R. Experience with the HET-CAM method in the routine testing of a broad variety of chemicals and formulations. Alternatives to Laboratory Animals. 2010;38(1):39–52. https://doi.org/10.1177/026119291003800109.
  5. Batista-Duharte A, Jorge Murillo G, Pérez UM, Tur EN, Portuondo DF, Martínez BT, et al. The Hen’s egg test on chorioal-lantoic membrane. International Journal of Toxicology. 2016;35(6):627–633. https://doi.org/10.1177/1091581816672187.
  6. Szuhanek CA, Watz CG, Avram Ș, Moacă EA, Mihali CV, Popa A, et al. Comparative toxicological in vitro and in ovo screening of different orthodontic implants currently used in dentistry. Materials. 2020;13(24):5690. https://doi.org/10.3390/ ma13245690.
  7. Kennedy DC, Coen B, Wheatley AM, McCullagh KJ. Microvascular experimentation in the chick chorioallantoic membrane as a model for screening angiogenic agents including from gene-modified cells. International Journal of Molecular Sciences. 2021;23(1):452. https://doi.org/10.3390/ijms23010452.
  8. Kabinet Ministriv Ukrainy. Postanova№ 65-2021-p. Ofitsiinyivisnyk Ukrainy. 2021.Availablefrom: https://zakon.rada.gov.ua/ laws/show/65-2021-%D0%BF#Text (In Ukrainian).
  9. BinJadeed H, Almudimeegh AM, Alomran SA, Alshathry AH. A case of contact allergic dermatitis to topical minoxidil. Cureus. 2021;13(1):e12510. https://doi.org/10.7759/cureus.12510.
  10. Hudz N, Kobylinska L, Dmytrukha N, Korytniuk R, Wieczorek PP. Biological and analytical studies of peritoneal dialysis solutions. The Ukrainian Biochemical Journal. 2018;90(2):34–44. https://doi.org/10.15407/ubj90.02.034.
  11. National Toxicology Program. ICCVAM-recommended test method protocols [Internet]. Research Triangle Park (NC): National Institute of Environmental Health Sciences; 2023. Available from: https://ntp.niehs.nih.gov/whatwestudy/niceatm/ resources-for-test-method-developers/protocols.
  12. Kuzminov BP, Turkina VA, Chemodurova NYe. Otsinka khimichnykh rechovyn na khorioalantoisnii membrani embriona kurky (test HET-CAM) [Methodological guidelines. Evaluation of irritation action of substances on the chorioallantoic membrane of the chick embryo (HET-CAM test)]. Lviv; 2023. 20 p. https://doi.org/10.13140/RG.2.2.32544.56322 (In Ukrainian).
  13. Winter G, Koch AB, Löffler J, Lindén M, Solbach C, Abaei A, et al. Multi-modal PET and MR imaging in the Hen’s egg test-chorioallantoic membrane (HET-CAM) model for initial in vivo testing of target-specific radioligands. Cancers. 2020;12(5):1248. https://doi.org/10.3390/cancers12051248.
  14. Budai P, Kormos É, Buda I, Somody G, Lehel J. Comparative evaluation of HET-CAM and ICE methods for objective assessment of ocular irritation caused by selected pesticide products. Toxicology in Vitro. 2021;74:105150. https://doi.org/10.1016/j.tiv.2021.105150.
View article PDF