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Using Voltage to Optimize Protein Growth. Collagen Fibrillogenesis. Mary Beth Bird Mentor: Ramana Pidaparti, PhD. Mechanical Engineering Department July 23, 2007. Goals of Project. To trigger self-assembly mechanisms in collagen
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Using Voltage to Optimize Protein Growth Collagen Fibrillogenesis Mary Beth Bird Mentor: Ramana Pidaparti, PhD. Mechanical Engineering Department July 23, 2007
Goals of Project • To trigger self-assembly mechanisms in collagen • To establish an experimental setup that will support fibrillogenesis • To monitor and alter variables in experiment to determine the best environmental conditions for fibrillogenesis • To replicate experimental data in computational modeling derived from research data
Background on Collagen • Most common form of protein in the human body • Triple helical molecule consisting of 3 alpha chains of amino acids • There are 13 different kinds of collagen, Type I – XIII • Cross-linking during fibrillogenesis causes unique properties amongst the different types • Collagen can be found in all connective tissues; ie. bones, tissues, cartilage, ….
The Original Idea • Use metal substrate as a voltage conductor coated with SDS (Sodium Dodecylsulfate) • Layer with a 1-2cm sq of collagen in an acidic solution • Apply voltage across metal conductor • Monitor growth formation using electron microscope • Variables: pH, time, voltage, concentration of collagen solution
Problems Encountered • Type of substrate • Determining collagen Type and form to be used • Use of SDS (Sodium Dodecylsulfate, ionic surfactant) • Use of Hydroxylapatite (mineral) • Application of voltage using Corrosion Data Program • Physical experimental setup • Ability to monitor pH and temperature • Method of examining collagen formation • Method of quantifying and understanding data • Ability to confirm presence of fibrils • Prevention of corrosion
Concerns • Corrosion • How can we prevent corrosion? • Alter voltage? Alter pH using NaOH? Alter substrate? • Substrate Surface • How can we improve the surface of the substrate to prevent false images? • Analyzing Images • How do we confirm presence of fibrils? • What is the best way to take images? • SEM, AFM, Optical Microscope
Future of the Project • Continue to refine methods to decrease the number of experimental errors • Control and alter experimental variables ie. pH, temperature, voltage • Form a better method to quantify and examine data • Expand project to include DNA
Acknowledgements Project Team Members: • Sumeet Lall, HSURP • Dr. Ramana Pidaparti Mechanical Engineering Dept., VCU • Dr. Gary Bowlin Biomedical Engineering Dept., VCU • Dr. Peter C. Moon School of Dentistry, VCU • Eddie McCumiskey, Grad Student NanoMan Lab Mechanical Engineering Dept., VCU • Dr. Stephen Fong’s Lab Chemical and Life Sciences Engineering Dept., VCU
References 1. Encyclopedia of Human Biology. 2nd ed. San Diego, CA: Academic Press; 1997. 2. Du C, Cui F, Zhang W, Feng Q, Zhu X, de Groot K. Formation of calcium phosphate/collagen composites through mineralization of collagen matrix. Journal of Biomedical Materials Research [serial online]. 2000;50:May 31, 2007. 3. Sini P, Denti A, Tira M, Balduini C. Role of decorin on in vitro fibrillogenesis of type I collagen. Glycoconjugate Journal. 1997;14:871-874.