A Study of Drug Interaction on Muscle Cell Viability and Response Jovan Deas

1. Fold or cut poster here Fold or cut poster here Fold or cut poster here Fold or cut poster here A Study of Drug Interaction on Muscle Cell Viability and Response Jovan Deas Introduction Materials and Methods Experimental Design Diagram Conclusions The aim of the research was to determine what effect mixing different classes of blood pressure medications with potassium ions has on muscle cells as well as muscle strands. The data was analyzed for statistical significance using a one way ANOVA analysis performed on each set of data. For the cell study, the ANOVA test consisting of individual treatments resulted in a p value= 0.148. The test consisting of mixed treatments resulted in a p value= 0.109 For the muscle tissue study, the ANOVA test consisting of individual treatments resulted in a p value less than .001. The test consisting of mixed treatments resulted in a p value= 0.990. For the cell study conducted the research hypothesis was supported because cell viability decreased significantly For the muscle tissue study the research hypothesis was rejected however the percent muscle contractions of mixed treatments was significantly lower than treatments that were independently applied. The implications on the data that was received shows that mixing several common classes of blood pressure medications with potassium ions which are commonly called simple salts, result in a significant decrease in muscle cells. This is important because if these medications are prescribed together, patients muscle activity can be affected. Sources of error include limitations of samples/ groups, possible pipetting errors in small volumes, and possible air contamination when conduction the muscle study. These possible sources of error may have caused a wide variation in repeated trials. As American diets increase in salt consumption and fat, a diagnosis of high blood pressure or hypertension becomes more common. High blood pressure itself is a risk factor for more serious cardio vascular diseases (Zieve, 2010). Recommendations of lifestyle changes, decrease in salt consumption and fat, along with an increase in activity and a prescription for high blood pressure medication are usually the norm. What if mixing two regularly used medications could greatly affect someone's health? The research problem is that other blood pressure medications classes along with ACE inhibitors may cause muscle cells to die as well as decrease muscle contraction when they are combined with potassium supplements. To test to see if these medications decrease cell viability and muscle contration when used with potassium supplements, the drugs were combined with potassium supplements to determine if they decrease or maintain the viability of the muscle cells and decrease the percent muscle contraction. This research has real world application because when these medications are mixed they may be harmful to people. • Materials • Growth media • Trypsin • 70 % ethanol • Gloves, goggles, lab coats • Autoclavable tubes 15 ml • Cell counter • Beaker for waste • Potassium supplements • Benazepril • Chlorthalidone • Nadalol • Muscle cells • Muscle tissue • Several steps were needed to complete the cell viability section of the experiment. One of the first steps was to acquire all materials, such as muscle cells and the various medications. The next step was to set up the well plates with the cells. The next step was to apply all of the treatments to the cell well plates. Before applying treatments, each treatment was first sterilized through a sterile .2 micron filter into sterile containers while working under the flow hood. After 24 hours of being incubated the cells should be attached to the bottom of the well plates. The well plates were then taken from the incubator and placed under the sterile hood. The sterilized treatments were then applied to the cells by using fresh pipette tips with every concentration and medication. The next step was to trypsinize the cells. The next step of culturing cells was to count the cells. Now the hemacytometer was placed under a microscope and all the living cells in the outer four quadrants were counted. That number was multiplied by 5 to get the total number of cells per ml. The final step of cell culturing was to treat and discard all materials. Data Analysis Hypothesis As the concentration of blood pressure medication and potassium ions increases, the muscle cell viability will decrease when compared to an untreated control group. If combined treatments of BP medications and potassium ions are added to the striated muscle samples, the muscle will lose its ability to respond to an electrical impulse and will not contract, or contract to a lesser degree when compared to the control. Figure 1: The graph above shows the average cell count of muscle cells given individual and mixed treatments. The bars represent each treatment and the controls mean cell count. The line shows an error line that is I standard deviation above and below the mean cell count. Future Research The delimitations of the experiment included testing a limited number of treatments, at one base concentration. To further this study, different concentrations of the medications can be applied to cells and muscle tissue to better observe trends. More trials can also be used to increase the validity of the statistical analysis. Background Information References The independent variables for this experiment were the type of the various blood pressure medications tested with and without adding potassium ions. The classes of blood pressure medications used in experimentation were ACE inhibitors, diuretics, and Beta blockers. The ACE inhibitor that was used was benazepril. The diuretic that was used was Chlorthalidone. The beta blocker used was Nadolol..There was also a control group that will receive no treatment. These medications are very important in helping maintain a stable blood pressure, but caution should be used when taking potassium. Potassium supplements can also assist the body in keeping healthy. The results in this project can further reveal any negative side effects to muscle tissue when taken in combination The dependent variable for experimentation is the cell viability of the muscle cells measured in number of viable cells per µl. In this experiment a combination of drugs are being tested to determine what effect the combinations have on the life of these cells. This was measured with a hemacytometer. The dependent variable for the second muscle tissue phase is percent muscle contraction. This was measured in cm. • Alper, A. (2005, June/ July). How do blood pressure medicines differ? . Kidney Beginnings: The Magazine, 4(2), • American Heart Association , . (2011, January 21).Types of blood pressure medications . Retrieved from http://www.heart.org/HEARTORG/Conditions/HighBloodPressure/Types-of-Blood-Pressure-Medications_UCM_303247_Article.j • Hodgkin, A.L. (1960). Potassium contractures in single muscle fibres. The Journal of Physiology, 153. Retrieved from http://jp.physoc.org/content/153/2/386.full.pdf • Jamerson, K., and Weber, M., (2008, December 4). Benazepril plus amlodipine or hydrochlorothiazide for hypertension in high-risk patients. Retrieved from http://content.nejm.org/cgi/content/abstract/359/23/2417 • Zieve, D. (2010, July 29). Hypertension . Retrieved from http://www.ncbi.nlm.nih.gov/pubmedhealth/PMH0001502/ Figure 2: The graph above shows the average percent contraction of muscle strands given individual and mixed treatments. The bars represent each treatment and the controls mean percent contraction. The line shows an error line that is I standard deviation above and below the mean percent contraction. Trends Cell study: • BPMs benazepril and nadalol, caused a slightly significant increase • Potassium ions caused a slightly significant increase • BPM chlorthalidone showed a significant decrease • All BPMs mixed with the potassium ions resulted in a significant decrease Contraction Study: • All individual treatments of BPMs showed a significant increase • Treatments with BPMs mixed with potassium ions showed a significant increase • Treatment of potassium ions caused a slight decrease Sterile tubes containing filtered treatments Well plates prepared under laminar flow hood Counting viable cells using a microscope and a hemacytometer