To design a Standard Operating Procedure that describes the procedure for the 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one (oxazolone) induced psoriasis model in experimental mice.
2.0 SCOPE
This SOP provides details on handling, randomization, oral dosing, and evaluation/calculation of anti-psoriatic activity.
3.0 RESPONSIBILITY
Chief scientists in charge, study directors, or study investigators shall be responsible for taking the above experimental procedure.
4.0 Procedure
4.1.1 Mice will be quarantined for 1 week as per the in-house SOP if required.
4.1.2 Mice will be kept in a thermally regulated animal house (24˚C±1˚C) with a relative humidity of 50% ±20% or renewed air supply or 12 hours dark/light cycle.
4.1.3 Mice will be acclimatized to the experiment room 3 days prior to the experimental procedure.
4.1.4 All mice will be randomized based on body weight into different groups and recorded.
4.1.5 The abdomen area of all mice will be clipped with an electrical clipper and remove out all hairs.
4.1.6 The mice's abdomen will be clipped (3cm X 5cm) to a sufficient working area.
4.1.7 1.5 % (w/v) of oxazolone will be prepared in acetone (vehicle) to sensitize the mice.
4.1.8 On day 0 all mice will be sensitized by 1.5% (w/v) of oxazolone by application on the clipped abdomen 100µl using a micropipette.
4.1.9 Mice will be monitored daily for clinical signs and symptoms and the body weight will be measured and recorded.
4.1.10 On day 7 the ear thickness of all mice will be measured using a digital vernier caliper before the challenge.
4.1.11 Then the mice will be challenged by 1.5% (w/v) of oxazolone by application on the right ear at 20µl/ear and the left ear will be treated with acetone (vehicle) only and served as control.
4.1.12 After the challenge the ear thickness will be measured using a digital vernier caliper at 6, 24, 48, and 72 hr.
4.1.13 Ear thickness will be measured in three areas of the ear that is upper ear margin (UEM), middle ear margin (MEM), and lower ear margin (LEM).
4.1.14 The increase in ear thickness will be determined by subtracting the ear thickness of day 0 (before challenge or acetone application) from the respective time point thickness.
5.0 REFERENCES
5.1.1 Luting Zeng, Yingqin Liu, Congcong Xing, Yijie Huang, Xin Sun, and Guangcheng Sun. Saponin from Periploca forrestii Schltr Mitigates OxazoloneInduced Atopic Dermatitis via Modulating Macrophage Activation. Mediators of Inflammation Volume 2020, Article ID 4346367, 13 pages https://doi.org/10.1155/2020/4346367.
5.1.2 Young Bok Lee, Su Jin Kim, Sae Mi Park, Kyung Ho Lee, Hyung Jin Han, Dong Soo Yu, So Youn Woo , Seong Taek Yun , Se-Yeong Hamm , Hong Jig Kim , Jin-Wou Kim. Immunomodulatory Effects of Deokgu Thermomineral Water Balneotherapy on Oxazolone-Induced Atopic Dermatitis Murine Model. Ann Dermatol Vol. 28, No. 2, 2016.
5.1.3 Sullim Lee, Hyun Jegal, Sim-Kyu Bong, Kyeong-No Yoon, No-June Park, Myoung-Sook Shin Min Hye Yang, Yong Kee Kim, and Su-Nam Kim. Anti-Atopic Effect of Acorn Shell Extract on Atopic dermatitis-like Lesions in Mice and Its Active phytochemicals. Biomolecules 2020, 10 (1), 57 https://doi.org/10.3390/biom10010057
5.1.4 Shishu Goindi, Gautam Kumar, and Amanpreet Kaur. Novel flexible vesicles based topical formulation of levocetirizine: in vivo evaluation using oxazolone-induced atopic dermatitis in the murine model. J Liposome Res, 2014; 24(3): 249–257.
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