model for the treatment of aneurysms

190 Pages·2017·4.18 MB·English

by  Lisa M. Reece

Purdue University
Purdue e-Pubs
Open Access Dissertations Theses and Dissertations
Fall 2014
Development of a static bioactive stent prototype
and dynamic aneurysm-on-a-chip(TM) model for
the treatment of aneurysms
Lisa M. Reece
Purdue University, Birck Nanotechnology Center
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Part of theBiomedical Engineering and Bioengineering Commons, and theNanoscience and
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Recommended Citation
Reece, Lisa M., "Development of a static bioactive stent prototype and dynamic aneurysm-on-a-chip(TM) model for the treatment of
aneurysms" (2014).Open Access Dissertations. 349.
https://docs.lib.purdue.edu/open_access_dissertations/349
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i
DEVELOPMENT OF A STATIC BIOACTIVE STENT PROTOTYPE AND
DYNAMIC ANEURYSM-ON-A-CHIP™ MODEL FOR THE TREATMENT OF
ANEURYSMS
A Dissertation
Submitted to the Faculty
of
Purdue University
Lisa M. Reece
In Partial Fulfillment of the
Requirements for the Degree
Doctor of Philosophy
December 2014
Purdue University
West Lafayette, Indiana
ii

To Ron, Autumn, and Ron Jr., for making my life so blessed with your forbearance, love,
understanding, and lots of humor and laughs! I could not have done this without your
devotion to me and our devotion to each other.
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ACKNOWLEDGEMENTS
I would like to acknowledge and thank my Lord and Savior, Jesus Christ, because
without Him I would not have been able to endure the many hours of hard work, study,
and fortitude required to get this far in my academic or personal life.

I wish to recognize my family members in Illinois, Oklahoma, Missouri, and Texas who
have prayed for me and have encouraged me. They never forgot to cheer me up and
make me feel loved and important. Thank you so much for all of the support through the
years.

To my dissertation committee: James F. Leary, Sophie Lelièvre, Sherry Voytik-Harbin,
and Steven Wereley, I offer my deepest thanks and gratitude for guiding me through a
very rewarding project and giving me helpful advice and showing faith in me. I want to
also thank Raji Sundararjan, who has helped me with prayers, love, enthusiasm, great
advice, and scientific counselling. I must express great appreciation to Jean Paul Allain
who was instrumental in obtaining funding for the initial project, and for allowing me to
be a Co-PI with him on the grant.

I want to thank the graduate students without whom I could not have performed a lot of
iv
the work in a timely manner: Sandra Arias, Moník Echeverry, Jian-Wei Khor, Fernando
Pastrana, Raviraj Thakur, and Emily Walker. Christy Cooper has always been a close
friend and colleague with whom it has been fun to hang out, share an office and enjoy
stimulating conversations, but I also want to say ‘thank you’ for all the chemistry help
she has provided me. I want to recognize Desirée White-Schenk for always helping me
out and for being a great sounding board when our mutual frustrations ran high.

Heartfelt sentiments go to Gloria Powell who helped me with the graduate school red
tape. In addition, I would like to express my gratitude to the BMS Department Chair:
Laurie Jaeger, who has gone to bat for me along with Dr. Leary on more than one
occasion. For this, I am extremely humbled and forever grateful.

I cannot continue without recognizing my Birck Nanotechnology Center family: all the
engineers, managers, and other staff (especially Nancy Black, Kenny Schwartz, Guy
Telesnicki, and Ira Young). You have made my academic and professional life all the
more enjoyable and bearable. I consider myself blessed to have worked beside such a
remarkable group of experienced, talented people. I also want to thank Mary Jo Totten
for the love, support and prayers she has given me especially through a particularly
difficult time in my life.

Finally, it is with increased satisfaction that I wish to express my thanks to my boss,
mentor, advisor, colleague and dear friend of many years, Dr. Leary. You have been a
great influence on my career and life and have helped to shape it with your skill,
v
intelligence, wit, verve, humor, and patience. You have been a huge inspiration to me
and it has been one of my greatest pleasures to have had the opportunity to work for you
and beside you these many years.
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TABLE OF CONTENTS
Page
LIST OF TABLES .............................................................................................................. x
ABSTRACT .................................................................................................................... xxv
CHAPTER 1. INTRODUCTION ................................................................................. 1
1.1 Rationale ................................................................................................................. 1
1.2 Specific Aims .......................................................................................................... 2
CHAPTER 2. BACKGROUND ................................................................................... 5
2.1 Definition of Aneurysms......................................................................................... 5
2.2 Common Treatments of Aneurysms ....................................................................... 7
2.2.1 Coiling Therapy to Occlude Aneurysms ..........................................................8
2.2.2 Clip Ligation as Alternative to Coiling ..........................................................12
2.2.3 Bioactive Stents for Aneurysm Treatment .....................................................14
2.3 Aneurysm-on-a-Chip™ Design ............................................................................ 17
2.4 Characteristics of Biomaterials ............................................................................. 19
2.5 PDMS – Choice Material for BioMEMS Device ................................................. 22
2.6 BNC – Choice Material for Bioactive Stent ......................................................... 25
2.7 HASMC – Ideal Cells for Bioactive Stent and AOC Prototypes.......................... 26
2.8 Various Assays to Identify Cellular Mechanisms ................................................. 27
2.9 Hemodynamics and Aneurysm Formation ........................................................... 30
CHAPTER 3. AIM 1: ENHANCE CELLS AND BACTERIAL
NANOCELLULOSE FOR ULTIMATE ATTRACTION TO A STATIC BIOACTIVE
STENT…………… .......................................................................................................... 32
3.1 Introduction ........................................................................................................... 32
3.2 Materials and Methods .......................................................................................... 33
3.2.1 Scheme 1: Magnetic Labeling of Cells and Apoptosis Testing ....................33
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Page
3.2.1.1 Magnetic Tagging of HASMC Via PU ................................................... 33
3.2.1.2 Electroporation of HASMC With SPION ............................................... 34
3.2.1.3 Determination of Iron Content in HASMC ............................................. 35
3.2.1.4 Measurement of Possible DNA Damage by CometAssay® ................... 37
3.2.2 Scheme 2: Synthesis of BNC/MBNC and Static Bioactive Stent Prototypes39
3.2.2.1 Preparation of Standard BNC .................................................................. 39
3.2.2.2 Preparation of Magnetic BNC ................................................................. 41
3.2.2.3 Preparation of Static Bioactive Stents ..................................................... 47
3.3 Results ................................................................................................................... 48
3.3.1 Scheme 1: Magnetic Labeling of Cells and Apoptosis Testing ....................48
3.3.1.1 Magnetic Tagging of HASMC Via PU ................................................... 48
3.3.1.2 Electroporation of HASMC with SPION ................................................ 49
3.3.1.3 Determination of Iron Content in HASMC ............................................. 52
3.3.1.4 Measurement of Possible DNA Damage by CometAssay® ................... 55
3.3.2 Scheme 2: Synthesis of BNC/MBNC and Static Bioactive Stent Prototypes58
3.3.2.1 Preparation of Standard BNC .................................................................. 58
3.3.2.2 Preparation of MBNC ............................................................................. 59
3.3.2.3 Preparation of Static Bioactive Stents ..................................................... 64
3.3.3 Brief Summary of Results ..............................................................................65
3.4 Discussion and Conclusions ................................................................................. 66
CHAPTER 4. AIM 2: IMPROVE AND PROMOTE TISSUE GROWTH AND
BIOCOMPATIBILITY UTILIZING BACTERIAL NANOCELLULOSE ..................... 69
4.1 Introduction ........................................................................................................... 69
4.2 Materials and Methods .......................................................................................... 70
4.2.1 Scheme 1: Cellular Growth and Migration Assays .......................................70
4.2.1.1 HASMC Proliferation Tests .................................................................... 70
4.2.1.2 Wound Healing Assays with IONP Uptake ............................................ 73
4.2.2 Apoptosis Assays ...........................................................................................77
4.2.2.1 Coating MBNC for Biocompatability ..................................................... 77