Alternative to animal studies

alternative to animal studies

1. EVALUATION SEMINAR ON ALTERNATIVE TO ANIMAL STUDIES PRESENTED BY: MR. UTTAM PAUL 1ST M. PHARM DEPT. OF PHARMACOLOGY FACILIATED TO: DR. SHIVALINGE GOWDA K.P. H.O.D, DEPARTMENT OF PHARMACOLOGY

2. TABLE OF CONTENTS   OVERVIEW DEFINITION TO ALTERNATIVE TECHNIQUES DIFFERENT TECHNIQUES TO BE USED AS ALTERNATIVES TO ANIMAL TESTING Full thickness skin model (in vitro method) In sillico methods Cell line technique. Patch clamp method REFERNCE

3. OVERVIEW

4. OVERVIEW Animal usage in scientific research and testing has raised controversy and criticism for long . A number of legislative initiatives have been proposed from time to time to limit animal research, or ensure proper treatment of animals . Animal welfare groups, or reformers, opposed various forms of animal research due to the increased use of animals for developing drugs and safety tests for pesticides.

5. ALTERNATIVE TECHNIQUES: DEFINITION: The term ‘alternative’ is used to refer to those techniques or methods that replace the use of laboratory animals altogether, reduce the numbers of animals required, or refine an existing procedure or technique to minimize the level of stress endured by the animal. However, the field of alternatives study particularly in vitro toxicology has evolved into a respected discipline and is attracting competent and motivated scientists around the world.

6. It is not possible to replace whole animal models with in vitro systems to evaluate drug effects on major organ systems. However, techniques can greatly reduce the number of animals needed, and refined protocols can improve the design efficiency and quality of studies, and lessen stress and discomfort experienced by lab animals. Generally instrumented animal models can be reused in major organ systems toxicology (MOST) for studies to evaluate more than one drug.

7. DIFFERENT TECHNIQUES TO BE USED AS ALTERNATIVES TO ANIMAL TESTING 1) Full thickness skin model (in vitro method) 2) In sillico methods 3) Cell line technique. 4) Patch clamp method.

8. 1) In-vitro methods C ell and tissue cultures are used to test product ingredients. Cell cultures are now also used routinely to test substances for mutagenic properties. Tissue cultures are additionally used to test substances for compatibility with mucous membranes.

9. Full Thickness Skin Model:

10. A test substance is applied to the Full Thickness Skin Model so that its effect on the skin tissue can be systematically evaluated. The substance, e.g. a cream formulation, is applied topically using a brush. This can be done several times over a period of at least nine days. In this way, the effect of the substance on the cell layers in the skin can be studied. The standardized production of the model, in combination with its special properties, makes it suitable for use as an in-vitro alternative to animal testing

11. 2. In-sillico methods Substances with similar chemical structures often have similar properties. In these cases, therefore, knowledge of the properties of a few representative substances is sufficient to be able to deduce the properties of a series of similar substances. By analogy, certain properties of these representative substances can also be assumed to be properties of the other substances in the series. The required calculations are performed using specially developed computer programs.

12. Only these selected substances will then have to be tested according to the legally prescribed test methods . It is anticipated that combinations of such calculations will make it possible to narrow down the number of substances to be tested.

13. 3) Cell line technique The term cell line refers to the propagation of culture after the first subculture. Once the primary culture is sub cultured, it becomes a cell line. A cell line derived by selection or cloning is referred to as cell strain. Types of cell lines : Finite cell line. Continuous cell line.

14. SERIAL NO PROPERTIES FINITE CELL LINE (with limited culture life spans) CONTINUOUS CELL LINE (unlimited life ) 1 Growth rate Slow Fast 2 Mode of growth Monolayer Suspension 3 Yield Low High 4 Transformation Normal Immortal, Tumorigenic 5 Cloning efficiency Low High 6 Serum requirement High Low 7 Markers Tissue specific Chromosomal, antigenic.

15. APPLICATIONS OF CELL LINE Screening of anti-cancer drugs Cell based bioassays To determine the cytotoxicity In vitro screening of several drugs Production of anti-viral vaccines Cancer research, which require the study of uncontrolled cell division in culture Cell fusion technique, Genetic manipulation Gene therapy, Recombinant DNA therapy Biotechnology , Molecular biology……etc.

16. PATCH CLAMP TECHNIQUE Patch clamp technique is a technique in electrophysiology that allows the study of individual ion channels in cells. The technique is used to study excitable cells such as neurons, muscle fibers and the beta cells of the pancreas . Use a pipette to pinch off a small region of membrane. Provides access to: the inside of the cell a small region of membrane

17. TYPES Of PATCH CLAMP : A. On-cell B. Inside Out C. Whole Cell D. Outside-Out

18. Applications of Patch Clamp Technique: For the evaluation of antiarrhythmic agents. In kidney cells. Used for isolated ventricular myocytes from Guinea pigs to study a cardio selective inhibition of the ATP sensitive potassium channel. To identify multiple types of calcium channels. To measure the effect of potassium channel openers. Used in the molecular biology. Voltage clamp studies on sodium channels. Used to investigate a wide range of electrophysiological cell properties. Measurement of cell membrane conductance.