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Invivo evaluation techniques
- 1. IN-VIVO EVALUATION TECHNIQUES Presented by: Anushma Chorsiya M.Pharm 16/MPH/DPSRU/2018
- 2. In vivo studies are those in which the effects of various biological activities are tested on whole, living organisms or cells, usually, including humans, and animals, as opposed to a tissue extract or dead organism. Animal testing and clinical trials are major elements of in vivo research. 2 INTRODUCTION
- 3. Facts • First refered in late 19th century by Louis Pasteur administered anthrax to sheep to show importance of vaccines with his germ theory. • Ivan Pavlov, conducted experiment on dogs to demonstrate condition regarding memory and repetitive tasks in late 19th century. • Breakthrough in 1922 when animal testing was allowed for insulin to be isolated from dogs. • 1930, mordern anaesthetics and antibiotics developed using animal testing. • Anticoagulant and kidney dialysis, both life saving treatments, were introduced following animal testing. • 1950s animal testing aiding in the development of vaccines. • Later quarter of 20th century, development of many potent cancer drugs and drugs for HIV. 3
- 6. “ Peptic ulcer most serious gastrointestinal diseases in the world. It comprises of hetrogeneous disorders, manifested due to break down in gastrointestinal mucosal lining bathed by acid and pepsin. 6
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- 9. Peptic Ulcer Models • Water-Immersion Stress • NSAIDs induced gastric ulcers • Ethanol- induced gastric ulcers • Acetic acid- induced gastric ulcers • Histamine- induced gastric ulcers •Ischemia-reperfusion induced gastric ulcers •Cysteamine- induced duodenal ulcers •Ferrous iron- ascorbic acid induced gastric ulcer 9 • Reserpine- induced gastric ulcers •Serotonin- induced gastric ulcers •Pylorus- ligated- induced peptic ulcers •Diethyldithiocarbamate- induced peptic ulcers •Metlylene blue- induced ulcer •Indomethacin- histamine induced duodenal ulcers •Acetic acid- H. pylori- induced ulcers Peptic Ulcers induced by physiological, pharmacological or surgical manipulation in several animal species, but most experiments carried out in rodents . Several models used for testing or evaluation are as follows;
- 10. Principle Leads to accumulation of gastric acid in the stomach leading to acute gastric ulceration. Pylorus Ligation Induced Ulcers 10 Water-Immersion Stress Induced Ulcers Principle Gastric lesions, significantly enhanced by exposure to water immersion in stress.
- 11. Principle Inhibition of endogenous prostaglandin production and consequent loss of gastric mucosal defence. NSAIDs Induced Mucosal Damage 11 Ethanol Induced Peptic Ulcers Principle Ethanol damages superficial epithelial layers and inhibits prostaglandin release.
- 12. Principle Produces penetrating ulcers in fundus area of rat. Ulcerated area in anterior and posterior wall are identical. Acetic Acid Induced Gastric Ulcers 12 Histamine Induced Peptic Ulcers Principle Histamine has potent acid stimulating and vasodilating capability leading to gastric ulcers.
- 13. Evaluation : • Ulcer Index = 10/x (x = Total mucosal area / total ulcerated area) • Intensity of ulcers 0 – Normal Stomach 1 – Superficial mucosal erosion 2 - Deep ulcers 3 - Penetrated or perforated ulcers • Contents of the stomach analyzed for : Volume, pH, Free acidity and total acidity, Mucin and prostaglandin level. 13
- 14. 14 SOURCE PART EXTRACT MODEL Aegle marmelos Fam. Rutaceae Leaves Aqueous extract Pylorus ligated gastric ulcers Allium sativum Fam. Liliaceae Bulb Bulb juice Cysteamine induced gastric ulcer Annona squamosa Fam. Annonaceae Leaves Aqueous extract Pylorus ligation and Ethanol induced gastric ulcer Bauhinia variegate Fam. Caesalpiniaceae Root Ethanolic and aqueous extract Pylorus ligation, ethanol, and aspirin induced gastric ulcers Careya arborea Fam. Myrtaceae Stem Bark Ethanolic extract Ethanol, Cold Restraint stress and Pylorus ligation induced peptic ulcers Carica papaya Fam. Caricaceae Seed Aqueous extract Ethanol induced gastric ulcers Ficus religiosa Fam. Urticaceae Leaves Hydro alcholic extract Absolute ethanol, aspirin and pylorus ligation induced gastric ulcers
- 16. Cancer are a large family of diseses that involve abnormal cell growth with the potential to invade or spread to other parts of the body. They form a subset of neoplasms. 16
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- 18. Anticancer Models The first in vivo test models used for screening anticancer compounds were developed back in mid-1960s at the NCI. These syngeneic mouse leukemia models were initially grown as ascites tumors in mice. 18
- 19. • DMBA induced mouse skin Papillomas • N-methyl, N-nitrosourea (MNU) induced rat Mammary gland Carcinogenesis • DMBA induced rat Mammary gland Carcinogenesis • MNU induced Tracheal Squamous cell carcinoma in Hamster • MNU induced Prostate cancer in Gerbils • N,N- Diethylnitrosamine (DEN) induced lung adenocarcinoma in Hamester • Azoxymethane (AOM) induced Aberrant Crypt Foci in Rat • Hollow Fiber technique • Use of Xenografts • Nude mouse model • New born rat model • Transgenic mouse model • DMBA induced oral cancer in Hamster • DMBA Sustained release suture technique • 3-Methylcholanthrene induced Fibrosarcoma tumors in Mouse • 3-Methylcholanthrene induced skin tumors in Mouse • Benzopyrene induced Forestomach tumors in Mouse • Hepatocellular carcinoma • High Fat diet induced NAFLD/NASH model in mouse • Pancreatic Cancer Models • Angiogenesis Assays 19 Chemically Induced Tumor Models Methods Involving Cell Line/ Tumor Pieces Implantation Other Models • 1,2- Dimethylhydralazine induced Colorectal Adenocarcinoma in Rat and Mouse • N-Butyl-N-(4-hydroxybutyl)- nitrosamine induced carcinoma in Mouse
- 21. “ Diabetes mellitus group of metabolic disorder, characterized by absolute lack of insulin. About 2.8% of global populations affected by diabetes mellitus. Experimentally diabetes mellitus is generally induced by chemical, surgical and genetic manipulation. 21
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- 24. Antidiabetes Model Chemical Induced • Alloxan induced • Streptozotocin induced • Dithizone induced • Monosodium glutamate induced • Insulin antibodies induced •Ferric nitrilotriacetate induced • Goldthioglucose obesed Virus Induced • D-Variant Encephalomyocarditis (EMC-D) • Coxsackie viruses Diabetic Obese Rodent Models • Kuo Kondo mouse • ob/ob mouse • db/db mouse • Zuker Diabetic Fatty (ZDF) rat • New Zealand Obese (NZO) mouse • Otsuka Long-Evans Tokushima Fatty (OLETF) rat • Nagoya-Shibata- Yasuda (NSY) mouse • M16 mouse Hormone Induced • Growth hormone induced • Corticosteroid induced Diabetic Non-Obese Rodent Models • Goto Kakizaki (GK) rat • Cohen diabetic rat • Spontaneously Diabetic Torii (SDT) rat Surgical Model • Full and partial pancreatectomy models • Genetically modified models 24
- 25. Principle Broad spectrum antibiotic, causing beta islet cell damage by free radical generation. Streptozotocin Induced Dibetes 25 Virus Induced Diabetes Principle Etiological agents producing diabetes mellitus by infecting and destroying beta cells of pancreas.
- 26. Principle Dexamethosone, a steroid possessing immunosuppression action, causing an autoimmune reaction in the islet and produces type 1 diabetes. Corticosteroid Induced Diabetes 26 Ob/Ob Mouse Principle The ob/ob mouse strain, have leptin deficiency because of the mutation in leptin gene leading to severe insulin resistance. The ob/ob mice exhibit rapid gain in body weight, insulin resistance and hyperinsulinemia occurs.
- 27. Principle A polygenic non obese model shows insulin resistance, normolipidemia and impaired insulin secretion. Goto-Kakizaki Rat 27 Surgical Models of Diabetic Mellitus Principle Used to induce diabetes with complete removal of pancreas.
- 28. 28 SOURCE PART EXTRACT MODEL Caesalpinia bonducella Seeds Aqueous and 50% ethanolic extract Streptozotocin diabetic rats Capparis decidua Fruit Aqueous extract Alloxan induced diabetic rat Coccinia indica Leaves Dried extract Alloxanized diabetic dog Momordica charantia Fruit, Seeds and tissues Ethanolic extract Streptozotocin diabetic rats Tinospora ccordifolia Root Aqueous extract Alloxan induced diabetic rat Aegle marmelos Leaves Aqueous extract Alloxan induced diabetic rat Azadirachta indica Whole plant Hydro alcholic extract Streptozotocin diabetic rats
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- 31. • Carbontetrachloride induced hepatotoxicity • Galactosamine induced hepatotoxicity • Thioacetamide induced hepatotoxicity • Alcohol induced hepatotoxicity • Paracetamol induced hepatotoxicity • NSAIDs induced hepatotoxicity • Glucocorticoids 31 Models
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- 36. CONCLUSION 36
- 37. TOP 10 ANIMAL TESTING COUNTRIES 37 USA CHINA JAPAN AUSTRALIA CANADA UK FRANCE GERMANY TAIWAN BRAZIL
- 38. 11.5 MILLIONS ANIMALS WERE USED IN EXPERIMENTS ACROSS EUROPE 38
- 40. ALTERNATIVES 40 1 2 3 4 5 6
- 41. REFERENCES
- 42. • Blatt, N. L., Mingaleeva, R. N., Khaiboullina, S. F., Kotlyar, A., Lombardi, V. C., & Rizvanov, A. A. (2013). In vivo screening models of anticancer drugs. Life Sci. J, 10(4), 1892. • Karthikeyan M, Balasubramanian T, Kumar P (2016). In-vivo Animal Models and In-vitro Techniques for Screening Antidiabetic Activity. J Develop Drugs. 5(2):1-6. • Dewangan, H., Tiwari, R. K., Sharma, V., Shukla, S. S., Satapathy, T., & Pandey, R. (2017). Past and Future of in-vitro and in-vivo Animal Models for Diabetes: A Review. INDIAN JOURNAL OF PHARMACEUTICAL EDUCATION AND RESEARCH, 51(4), S522-S530. • Adinortey, M. B., Ansah, C., Galyuon, I., & Nyarko, A. (2013). In vivo models used for evaluation of potential antigastroduodenal ulcer agents. Ulcers, 2013. 42
- 43. • Kumar, A. E., Susmitha, K., Swathy, B., Ramu, E., & Venkatesh, B. (2014). A review on liver disorders and screening models of hepatoprotective agents. Int. J. Allied Med. Sci. Clin. Res, 2(2), 136-150. • Delgado-Montemayor, C., Cordero-Pérez, P., Salazar-Aranda, R., & Waksman-Minsky, N. (2015). Models of hepatoprotective activity assessment. Medicina universitaria, 17(69), 222-228. • Mushtaq, S., Daş, Y. K., & Aksoy, A. (2018). Alternative Methods to Animal Experiments. Türkiye Klinikleri. Tip Bilimleri Dergisi, 38(2), 161-170. • Modak, M., Dixit, P., Londhe, J., Ghaskadbi, S., & Devasagayam, T. P. A. (2007). Recent advances in Indian herbal drug research guest editor: Thomas Paul Asir Devasagayam Indian herbs and herbal drugs used for the treatment of diabetes. Journal of clinical biochemistry and nutrition, 40(3), 163-173. 43
- 44. 44 Thank YOU ! Any questions?