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Ammi Visnaga) PLANT EXTRACT on in VITRO CRYSTALLIZATION of CALCIUM OXALATE MONOHYDRATE

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Ammi Visnaga) PLANT EXTRACT on in VITRO CRYSTALLIZATION of CALCIUM OXALATE MONOHYDRATE EFFECT OF KHELLA (Ammi visnaga) PLANT EXTRACT ON IN VITRO CRYSTALLIZATION OF CALCIUM OXALATE MONOHYDRATE By SAIJIT DAOSUKHO A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2007 1 © 2007 Saijit Daosukho 2 To my Mom and Dad 3 ACKNOWLEDGMENTS I would like to thank everyone who supported me during my years in graduate studies. I am very grateful to my advisor, Dr. Hassan El-Shall, who provided the greatest help, support, and understanding through all of difficulties during my whole graduated study. Without his encouragement and guidance, I would have a totally different life. I thank my other advisor, Dr. Spyros Svoronos who guided me through my transition in chemical engineering PhD program. Moreover specials thanks to my external committee member, Dr. Saeed Khan, for his help explaining biological matters regarding kidney stone subjects. I would also like to thank other members of my committee, Drs. Richard Dickinson and Yider Tseng in the Department of Chemical Engineering. I am grateful for the help on plant extraction and HPLC experiments from Dr. Butterweck and her group. I would like to thank Dr. Yakov Rabinovich for his help with AFM measurement and interpretation, and a hearty thanks to Dr. Khan’s group, specifically, Karen Byer and Pat Glenton, and the PERC staff. I am appreciated the help and discussion from my fellow graduate students in Dr. El- Shall’s group, Dr. Stephen Tedeschi, Rhye Hamey, and Kerri-Ann Hue . I thank my good friends, Dr. Sukitti and Pawinee Punak, Theerapat and Pipawin Leesamphand, Dr. Kornvika Pimukmanaskit, Noppun Wongkittikraiwan and the Sartinoranont family for their help and mental support through out my years at the University of Florida. Hearty thanks to Shirley Kelly, Cynthia Sain, and Deborah Sandoval at the Chemical Engineering Department, who have provided superb help getting me through all the paper work. Special thanks to my families, who have always been the greatest supporter with love, care and encouraging words. Finally, I am grateful to the Particle Engineering Research Center and the Industrial Partners of the ERC and the Thai Government for financially supporting this research 4 TABLE OF CONTENTS Page ACKNOWLEDGMENTS ...............................................................................................................4 LIST OF TABLES...........................................................................................................................8 LIST OF FIGURES .........................................................................................................................9 ABSTRACT...................................................................................................................................14 CHAPTER 1 INTRODUCTION ..................................................................................................................17 2 LITERATURE SURVEY.......................................................................................................19 Kidney Stone Disease.............................................................................................................19 Urinary Tract and Kidney Structure .......................................................................................19 Composition and Structure of Kidney Stones ........................................................................23 Urine Composition and Its Role in Kidney Stone Formation.................................................25 Crystallization of Calcium Oxalate in Biological Systems ....................................................26 Nucleation Theory...........................................................................................................26 Crystal Growth ................................................................................................................30 Crystal Aggregation and Dispersion ...............................................................................31 Crystal-crystal interaction ........................................................................................31 Crystal-cell interaction .............................................................................................34 Treatment of Kidney Stone.....................................................................................................38 Extracorporeal Shockwave Lithotripsy ...........................................................................38 Oral Supplements ............................................................................................................39 Significance of Herbal Medicinal Treatment of Kidney Stone ..............................................40 Herbs Related to Kidney Stone Treatment......................................................................40 Ammi visnaga (Khella) Extract .......................................................................................41 Characterization Techniques ..................................................................................................43 High Performance Liquid Chromatography (HPLC)......................................................43 Inductively Coupled Plasma (ICP)..................................................................................44 UV-Vis Spectrometer ......................................................................................................44 Atomic Force Microscopy (AFM)...................................................................................45 3 MATERIALS AND METHODS ...........................................................................................47 Extract Preparation .................................................................................................................47 Extract Characterization .........................................................................................................47 Thin Liquid Chromatography (TLC)...............................................................................47 High Performance Liquid Chromatography (HPLC)......................................................48 Inductively Coupled Plasma (ICP)..................................................................................49 5 Oxalate identification ......................................................................................................50 Nucleation Study ....................................................................................................................50 Induction Time Measurement..........................................................................................50 Membrane Isolation.........................................................................................................51 Solution Preparation ........................................................................................................53 Calcium Titration Measurement......................................................................................54 Crystal Morphology and Crystallinity....................................................................................54 Crystal-Cell Interaction Study ................................................................................................55 Preparation of COM Crystals for AFM Study ................................................................55 Artificial Urine Solution Preparation ..............................................................................56 Cell Culture .....................................................................................................................57 Direct Force Measurement between COM Crystal and Kidney Renal Cells Using AFM.............................................................................................................................58 4 RESULTS AND DISCUSSION.............................................................................................60 Khella Extract Active Component Characterization ..............................................................60 Inorganic Component Identification................................................................................60 Organic Component Identification..................................................................................60 Thin liquid chromatiography (TLC) ........................................................................61 High performance liquid chromatography (HPLC) .................................................61 Effect of Khella Extract on Calcium Oxalate Crystallization ................................................64 Effect of Khella Extract on Nucleation of Calcium Oxalate Crystals.............................64 Light absorption measurement .................................................................................64 Calcium titration measurement ................................................................................67 Crystal morphology studies......................................................................................68 Surface energy and Gibbs free energy calculation...................................................73 Cell-retention of calcium oxalate crystal .................................................................81 Effect of Khella Extract Components on Calcium Oxalate Crystallization ...........................83 Effect of Khellin and Visnagin on Calcium Oxalate Crystallization ..............................84 Nucleation studies ....................................................................................................84
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