Anticancer Assessments for Skin Carcinoma: An In-Vivo Study
1.0
TEST SYSTEM DETAILS:
Species : Wistar Rat (Rat)
Age : 12-16 weeks
Sex : Male
No. of animals : 8/Group
Total animals : 40
2.0
ALLOCATION OF GROUPS:
Animal study:
|
GROUPS |
TREATMENT |
ANIMAL
NO. |
|
1 |
Normal Control (Vehicle Control) |
8 |
|
2 |
Disease Control, Carcinogen (Cancer
induced untreated animals) |
8 |
|
3 |
Carcinogen + Hydrogel contain Plant
extract 10 mg (LD) |
8 |
|
4 |
Carcinogen + Plant extract 30 mg (MD) |
8 |
|
5 |
Carcinogen + Plant extract 90 mg
(HD) |
8 |
3.0
METHOD:
Healthy animals will be selected, randomized based on body weight and divided into 5 different groups consisting of 8 animals each for anticancer activity. Animals will be divided into 5 groups of 8 animals in each. Group 1 animals served as normal control, while group 2 animals received DMBA alone 25 μg in 100 μL acetone. Animals in groups 3-5 received DMBA and herbal formulation (HF) at 10, 20, and 30 mg/kg body weight (b.w) in 1% DMSO respectively. HF will be administered orally daily a week, starting from one week before DMBA painting, to 25th week. At the end of the 25th week, the experiment was terminated. All animals will be euthanized, and immediately skin and liver tissues were dissected out and blotted dry before weighing.
Tumor induction and
assessment
The posterior side of each mouse was shaved with hair depilatory cream, left untreated for two days. Animals having no hair growth after two days were selected for the study. For tumor induction, the DMBA was applied topically, and monitored for 8 weeks. Body weight was recorded for each animal in all groups at weekly intervals, throughout the experimental period. At the termination of the experiment, all the animals were sacrificed by cervical dislocation. The skin tumor burden was calculated by multiplying the tumor volume and number of tumors/mice. The tumor volume was calculated by the formula, ν= (4/3) π [D1/2] [D2/2] [D3/2] where D1, D2 and D3 are the three diameters (mm3) of the tumors.
ENDPOINT PARAMETER(S):
Morphological analysis: observations including body weight, percentage of survival of animals, percentage of animals with tumor growth, tumor volume etc.
Histological Studies: it is a very significant method for investigation of the changes in cellular structure and an essential component of research.
Immunohistochemistry: IHC analysis of the animal tissues for Ki-67 and PCNA tumor markers will be carried out.
Biochemical analysis: Blood Samples and tissue homogenates will be utilized for biochemical evaluations like lipid peroxidation, antioxidant [Catalase, Superoxide Dismutase, Glutathione (GSH & GPx)] estimations etc.
ELISA: Estimation of Inflammatory Cytokines (TNF-α, IL-1β, IL-6) mainly involved in progression of skin carcinoma.
SAFETY
GUIDELINES
Ø Carcinogen-treated animals
will be kept separately on an identified slab or platform in the animal house
and will be monitored daily.
Ø Cages will be
well-labelled and instructions will be clearly mentioned.
Ø The carcinogen is not
spreaded by air and it gets absorbed and metabolized rapidly in the body of
animals.
Ø Cross-contamination of
cancer is not possible as the cancer is not growing to metastatic stage.
Ø 2 groups of induced animals
will be treated at a time to reduce the number of carcinogenic animals in a
particular time interval which further reduce the risk of contamination.
REFERENCE(S):
1. Schwarz, M., Münzel, P. A., & Braeuning, A. (2013). Non-melanoma skin cancer in mouse and man. Archives of toxicology, 87(5), 783–798. https://doi.org/10.1007/s00204-012-0998-9
2. Thamizharasi Gopalakrishnan, Sindhu Ganapathy, Veeravarmal Veeran, Nalini Namasivayam, Preventive effect of D-carvone during DMBA induced mouse skin tumorigenesis by modulating xenobiotic metabolism and induction of apoptotic events, Biomedicine & Pharmacotherapy, Volume 111, 2019, Pages 178-187, https://doi.org/10.1016/j.biopha.2018.12.071
3. Zhang, C. L., Zeng, T., Zhao, X. L., Yu, L. H., Zhu, Z. P., & Xie, K. Q. (2012). Protective effects of garlic oil on hepatocarcinoma induced by N-nitrosodiethylamine in rats. International journal of biological sciences, 8(3), 363–374. https://doi.org/10.7150/ijbs.3796
4. Mathur, S., Kaur, P., Sharma, M., Katyal, A., Singh, B., Tiwari, M., & Chandra, R. (2004). The treatment of skin carcinoma, induced by UV B radiation, using 1-oxo-5beta, 6beta-epoxy-witha-2-enolide, isolated from the roots of Withania somnifera, in a rat model. Phytomedicine: international journal of phytotherapy and phytopharmacology, 11(5), 452–460. https://doi.org/10.1016/j.phymed.2003.05.004
5. Sekizawa, J., Yasuhara, K., Suyama, Y., Yamanaka, S., Tobe, M., & Nishimura, M. (1994). A simple method for screening assessment of skin and eye irritation. The Journal of toxicological sciences, 19(1), 25–35. https://doi.org/10.2131/jts.19.25
6. de Oliveira, N. F. P., de Souza, B. F., & de Castro Coêlho, M. (2020). UV Radiation and Its Relation to DNA Methylation in Epidermal Cells: A Review. Epigenomes, 4(4), 23. https://doi.org/10.3390/epigenomes4040023
END OF THE DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the Animal House Facility
Hi. I’m Writer of Researchsop.com. ’ ’ Please share these SOPs to all concern pharma people for their development. I like to fullfill the need of curious people. These things inspire me to make things looks better.
0 comments:
Post a Comment