Acoustic Wave Biosensors for Biomechanical and Biological Characterization of Cells

ACOUSTIC WAVE BIOSENSORS FOR BIOMECHANICAL AND BIOLOGICAL CHARACTERIZATION OF CELLS by Huiyan Wu B.E., Xi’an Jiaotong University, P. R. China, 2008 M.E., Xi’an Jiaotong University, P. R. China, 2011 Submitted to the Graduate Faculty of Swanson School of Engineering in partial fulfillment of the requirements for the degree of Doctor of Philosophy University of Pittsburgh 2017 UNIVERSITY OF PITTSBURGH SWANSON SCHOOL OF ENGINEERING This dissertation was presented by Huiyan Wu It was defended on June 22, 2017 and approved by Dissertation Co-Advisor: James H.-C. Wang, Ph.D., Professor, Departments of Orthopaedic Surgery & Mechanical Engineering and Materials Science William S. Slaughter, Ph.D., Associate Professor, Department of Mechanical Engineering and Materials Science Patrick Smolinski, Ph.D., Associate Professor, Department of Mechanical Engineering and Materials Science Dissertation Advisor: Qing-Ming Wang, Ph.D., Professor, Department of Mechanical Engineering and Materials Science ii Copyright © by Huiyan Wu 2017 iii ACOUSTIC WAVE BIOSENSORS FOR BIOMECHANICAL AND BIOLOGICAL CHARACTERIZATION OF CELLS Huiyan Wu, PhD University of Pittsburgh, 2017 During past decades, interest in development of cell-based biosensors has increased considerably. In this study, two kinds of acoustic wave sensors are adopted as the cell-based biosensors to investigate the biomechanical and biological behaviors of cells, the quartz thickness shear mode (TSM) resonator and Love wave sensor. For the first part, the quartz TSM resonator is applied to detect the structural and mechanical properties of tendon stem/progenitor cells (TSCs), which are one kind of newly discovered adult cells in tendons, and the platelets from blood. Through the TSM resonator, the related viscoelastic properties of cells are extracted, which could indicate the state of cells in different physiological conditions. The TSM resonator sensor is utilized to characterize the aging-related viscoelasticity differences between the aging and young TSCs, and also to monitor the dynamic activation process of platelets. For the second part, a 36˚ YX-LiTaO3 Love wave sensor with a parylene-C wave guiding layer is proposed as a cell-based biosensor. A theoretical model is derived, to describe the Love wave propagation in the wave guiding layer, the adherent cell layer, and penetration into the liquid medium. The Love wave sensor is used to monitor the adhesion process of cells. Compared with TSM resonator, the response of Love wave sensor to the cell adhesion is primarily induced by the formation of bonds between cells and the substrate. The numerical results indicate that the adherent cell layer of iv various storage or loss shear modulus in certain range can cause evident, characteristic variations in propagation velocity and propagation loss, revealing the potential of Love wave sensors in providing useful quantitative measures on cellular mechanical properties. In addition, a Love wave sensor with a phononic wave guiding layer is introduced for non-operation signal filtering and sensor sensitivity improvement. Both two kinds of acoustic wave sensors present their own advantages as the cell-based biosensors, indicating advisable techniques for investigating cell biology in general and certain physiological processes in particular. v TABLE OF CONTENTS PREFACE ................................................................................................................................. XIX 1.0 INTRODUCTION ........................................................................................................ 1 1.1 MOTIVATIONS AND OBJECTIVES .............................................................. 3 1.1.1 Cell-based Biosensors ...................................................................................... 3 1.1.2 Cell-based Acoustic Wave Biosensors ............................................................ 6 1.2 SPECIFIC TASKS............................................................................................. 10 1.2.1 Task I: Aging-related Viscoelasticity Characterization of Tendon Stem/Progenitor Cells by a Thickness Shear Mode Resonator ................. 10 1.2.2 Task II: Real-time Monitoring the Activation Process of Platelets Using a Thickness Shear Mode Resonator ................................................................ 11 1.2.3 Task III: A Cell-based Love-mode Acoustic Wave Biosensor with a Viscoelastic Wave Guiding Layer ................................................................. 12 1.2.4 Task IV: A Sensitivity-enhanced Love-mode Acoustic Wave Biosensor with a Phononic Wave Guiding Layer ......................................................... 13 2.0 BACKGROUND ........................................................................................................ 15 2.1 PIEZOELECTRICITY ..................................................................................... 15 2.1.1 Piezoelectric Effect ......................................................................................... 16 2.1.2 Piezoelectric Materials ................................................................................... 18 2.1.3 Equivalent Circuit Method for Piezoelectric Transducers ........................ 22 2.1.4 Piezoelectric Devices ...................................................................................... 27 2.2 ACOUSTIC WAVE SENSORS ........................................................................ 29 vi 2.2.1 Vibration in Piezoelectric Substrate ............................................................. 30 2.2.2 Classification and Comparison of Acoustic Wave Sensors ........................ 38 2.2.3 Applications of Acoustic Wave Sensors ....................................................... 41 2.2.4 Research Progress in Acoustic Wave Biosensors ........................................ 43 2.3 THICKNESS SHEAR MODE RESONATORS ............................................. 45 2.3.1 Transmission Line Model for Thickness Shear Mode Resonators ............ 47 2.3.2 Extraction of Complex Shear Modulus of Adherent Cells ......................... 51 2.4 LOVE-MODE ACOUSTIC WAVE SENSORS ............................................. 54 2.4.1 Love Wave Propagation in a Layered Love Wave Sensor ......................... 55 2.4.2 Determination of Propagation Velocity and Loss in a Layered Love Wave Sensor .............................................................................................................. 61 3.0 AGING-RELATED VISCOELASTICITY CHARACTERIZATION OF TENDON STEM/PROGENITOR CELLS BY A THICKNESS SHEAR MODE RESONATOR ............................................................................................................ 65 3.1 INTRODUCTION ............................................................................................. 65 3.1.1 Tendon Stem/Progenitor Cells ...................................................................... 65 3.1.2 Viscoelasticity Characterization of Cells ..................................................... 68 3.2 MATERIAL PREPARATION AND EXPERIMENTAL DETAILS ........... 69 3.2.1 Isolation and Culture of Tendon Stem/Progenitor Cells ............................ 69 3.2.2 Measurement and Characterization of Tendon Stem/Progenitor Cells.... 70 3.3 THEORETICAL MODEL AND ANALYTICAL METHODS .................... 72 3.4 RESULTS AND DISCSSIONS ......................................................................... 73 3.4.1 Tendon Stem/Progenitor Cells Adhesion on the Substrate ........................ 73 3.4.2 Viscoelasticity of Tendon Stem/Progenitor Cells ........................................ 77 3.4.3 Cytoskeleton of Tendon Stem/Progenitor Cells .......................................... 80 3.4.4 Comparison of TSM Resonator with Other Measurement Methods ........ 81 vii 3.5 CONCLUSIONS ................................................................................................ 85 4.0 REAL-TIME MONITORING THE ACTIVATION PROCESS OF PLATELETS USING A THICKNESS SHEAR MODE RESONATOR ...................................... 86 4.1 INTRODUCTION ............................................................................................. 86 4.1.1 Platelets and Platelet-rich Plasma ................................................................ 86 4.1.2 Detection of Platelet Activation..................................................................... 89 4.2 MATERIAL PREPARATION AND EXPERIMENTAL DETAILS ........... 91 4.2.1 Isolation and Washing of Platelets ............................................................... 91 4.2.2 Measurement and Characterization of Platelets ......................................... 91 4.3 THEORETICAL MODEL AND ANALYTICAL METHODS .................... 93 4.4 RESULTS AND DISCSSIONS ......................................................................... 94 4.4.1 Platelet Adhesion and Activation .................................................................. 94 4.4.2 Viscoelasticity and Cytoskeleton of Platelets ............................................... 97 4.4.3 Monitoring Activation Process of Platelets in Different Concentrations 104 4.4.4 Comparison of TSM Resonator with Other Measurement Methods ...... 109 4.5 CONCLUSIONS .............................................................................................. 110 5.0 A CELL-BASED LOVE-MODE ACOUSTIC WAVE BIOSENSOR WITH A BIOCOMPATIBLE WAVE GUIDING LAYER .................................................. 112 5.1 INTRODUCTION ..........................................................................................

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