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A novel ultrasonic stimulation system to enhance cell viability using piezoelectric micromachined ultrasonic transducers (pMUTs) with transwells

A novel ultrasonic stimulation system to enhance cell viability using piezoelectric micromachined ultrasonic transducers (pMUTs) with transwells
Translated Title
pMUT 과 transwell 을 이용한 세포의 활동성 증진을 위한 초음파 세포 자극 시스템의 개발
Lee, Won Jun
DGIST Authors
Lee, Won Jun; Choi, Hong Soo; Choi, Ji Woong
Choi, Hong Soo
Choi, Ji Woong
Issue Date
Available Date
Degree Date
2015. 2
Piezoelectric micromachined ultrasonic transducer (pMUT)Cell stimulation system12-well transwellPC 12 cellProliferation세포 자극 시스템12 웰 트랜스웰PC12 세포세포분열
A new ultrasonic cell stimulation system was designed and fabricated using 10 by 10 and 10 by 29 piezoelectric micromachined ultrasonic transducer (pMUT) arrays. The sizes of pMUT arrays are 2.5 by 2.5 mm and 2.27 by 6.84 mm. The diameter and pitch of pMUT arrays are 120 µm and the measured resonance frequency of each single pMUT element was 1.494 MHz in water. The acoustic peak pressures are 0.22 MPa and 0.18 MPa for 10 by 10 pMUT and 10 by 29 pMUT, respectively. The spatial average temporal average (SATA) intensity are 612.8 ± 183 mW/cm2 and 200 ± 22 mW/cm2 which are converted by the acoustic pressure of 10 by 10 and 10 by 29 pMUT array. The fabricated pMUT arrays were attached on print circuit boards (PCBs) and wire bonding was done for electrical connection. Each PCB and pMUT array was coated with parylene C for biocompatibility and waterproofing. For cell culturing, a twelve-well transwell was assembled one a PCB with ten pMUT arrays by epoxy. After curing the epoxy, polydimethylsiloxane (PDMS) was coated on the epoxy to enhance biocompatibility of the ultrasonic cell stimulation system. The size of the cell stimulation system is 13.4 ☓ 17.3 ☓ 2.5 cm and it is small enough to place the whole system in an incubator. The fabricated ultrasonic cell stimulation system was used for cell stimulation to characterize the effect of ultrasonic cell stimulation for proliferation of PC12 cells. Ultrasonic cell stimulations were performed for 16 groups in combination of stimulation time for 5 min, 10 min, 20 min, and 30 min and duty cycle for 15 %, 30 %, 50 % and 70 %. There was also a control group without ultrasonic stimulation. All of the stimulated groups showed positive effect on cell proliferation. For the most significant effect, the 5 min stimulation group showed more increased number of the cells as 238 %. Furthermore, the proliferation pattern was different along the stimulation time, not duty cycle. As a result, short stimulation time and long stimulation time occurred different tendency. Short stimulation groups showed 118 %, 122 %, and 134 % increase rate of cell number during stimulation, however long stimulation groups showed 117 %, 105 %, and 152 % increase rate of cell number during stimulation. Long time stimulation group has different increase pattern after last stimulation. In this thesis, new ultrasound cell stimulation system using the pMUT was developed for overcoming the limitation of conventional cell stimulation system using bulk ceramic transducer. After cell stimulation for PC12 cells, all stimulation group has averagely 150 % increase rate of cell number than control group. ⓒ 2015 DGIST
Table Of Contents
1. INTRODUCTION 8-- 1.1 Background 8-- 1.2 Necessity of cell stimulation using ultrasound 9-- 1.3 Research trend or Related research works 12-- 1.3.1 Cell stimulation sytem with bulk transducer 12-- 1.3.2 Capacitive micromachined ultrasonic transducers (cMUT) 13-- 1.4 Objective of research 15-- 2. DESIGN AND FABRICATIONS 17-- 2.1 Cell stimulation system design 17-- 2.2 Fabrication Process 20-- 2.2.1 Fabrication process for 2-D pMUT array 20-- 2.2.2 Assemble the 2-D pMUT array and 12-well transwell 24-- 3. CHARACTERIZATION AND EXPERIMENT SETUP 28-- 3.1 Impedance analyze of the 2-D pMUT array 28-- 3.2 Intensity of the 2-D pMUT array 30-- 3.3 Cell stimulation setup 33-- 3.3.1 Cell stimulation plan 33-- 3.3.2 Cell culture 34-- 3.3.3 Cell statistical analysis 34-- 3.3.4 Immunocytochemistry 34-- 4. RESULTS AND DISCUSSIONS 36-- 4.1 Electrical characterization of the pMUT array 36-- 4.2 Acoustic characterization 37-- 4.3 Results of the cell stimulation 43-- 4.4 Discussion 54-- 5. CONCLUSIONS 56-- REFERENCES 58-- APPENDIX 65
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