Psoriasis Animal Study Background:
1. Psoriasis - characterized by itching. redness, thickness, scaling, pathologically it causes Inflammation (Subdermal region), Parakeratosis (Nuncleated cells in Stratum Corneum), Neovascularization (formation of new blood vessels), Epidermal hyperplasia, Induration (Thickness), Erythema (Redness).
2. TPA-induced Psoriasis model is preferred to get results for the treatment of psoriasis-like lesions. This model does not give complete information on whether the test compound is completely effective for the treatment of psoriasis or not. Parakeratosis is not found in TPA induced Psoriasis (Nucleated Cells)
3. In Imiquimod-induced psoriasis-like skin inflammation, this experiment shows a high level of inflammation while inducing in mice model, generally, in psoriasis, the inflammation does not occur to such extent.
4. Several mouse models of psoriasis including chemical-induced models (topical application of imiquimod to the skin) and genetically engineered mice (epidermal deletion of JunB/c-Jun, constitutive activation of epidermal STAT3, and epidermal overexpression of Tie2) have been used to study the pathophysiology of the disease; but such models cannot fully summarize all molecular and cellular pathways occurring in human psoriasis. In human Xenotransplantation, the pre-psoriatic skin onto immunodeficient mice and initiating its conversion into a psoriatic plaque is the best model to separate the mechanisms occurring during the progress of human psoriasis.
5. Treatment: The latest highly effective treatment is the use of monoclonal antibody; Ustekinumam, which neutralized P17-40 alpha and beta subunits of inflammatory cytokines (IL-17, IL-12, IL-23).
7. TPA is also sold
under the name of Phorbol 12-mystic 13-acetate Powder (PMA), while you
purchase, you will get it in amber color, around 2 ml bottle size, that
contains nothing, while you try to see inside the bottle. It is mainly coated
inside the bottle's bottom part.
8. TPA Inducing solution is prepared by preparing 2 ml stock solution by adding 2 ml of acetone. While you use the TPA for induction purposes, inject out the required quantity is supposed 200 micro-liters, then add more exact 600 micro-liters of acetone to prepare the final inducing solution. In the chronic model, inject out the required TPA solution in small quantities, and the remaining amount stored at -20-degree temperature.
10. In the psoriasis study
plan, after 1 hr of TPA induction, the topical application of the drug should be done and the thickness of the ear should be measured after 4hrs of dosing using Vernier caliper.
11. For necropsy use an ear punch of 5 inches to remove the TPA-induced ear parts (two punch cuts) and two from the control ear, weight and keep the one portion in LN2 and another one in Formalin from both ears, the last remaining part of both whole ears keep in the LN2. It means separating two punches from each ear for further in-vitro and histopathology procedures.
12. In this experiment we can use Evans blue dye, generally, it is used to capture pictures of the stained ear, it will show the difference between the control and TPA-induced ear. (leakage of Evans blue dye from the circulation is used to monitor extravasation of the ear skin of CD1 mice. 3% Evans blue dye in saline at a dose of 300 mg/kg body weight is used for intraperitoneal injection in mice for 4 h after TPA administration.
13. Before the necropsy collect blood through an orbital route and centrifuge it to separate blood serum at 5000 xG for 15 minutes. During necropsy collect two ear punches from each ear of 5 mm round shape, one for histopathology keep it 2 ml Eppendorf suspending in NBF, and the second one for biochemistry keep it in keep it 2 ml Eppendorf preserve it LN2. The remaining portion of both ears preserves in LN2 for other in-vitro experiments. Dissect the animals and remove the spleens and livers of all animals and weigh them and preserved them in NBF, for the evolution of histopathological changes.
14. Before conducting psoriasis experiments on animals you should confirm whether the product will give the results or whether the TPA disease induction is effective or not, for that you should try the acute psoriasis model. In this model apply the TPA in three animals and check its effect for 2, 4, 6, 8, 12, 16, and 24 hrs. By this method, you can solve two issues, Firstly, what is the timing when TPA gives the best results Second, if when your product reduces more than 30 % of diseases condition then it might be an effective treatment for psoriasis.
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