Wright State University CORE Scholar Browse all Theses and Dissertations Theses and Dissertations 2018 Coupling of Mechanical and Electromagnetic Fields Stimulation for Bone Tissue Engineering Alyaa I. Aldebs Wright State University Follow this and additional works at: https://corescholar.libraries.wright.edu/etd_all Part of the Biomedical Engineering and Bioengineering Commons Repository Citation Aldebs, Alyaa I., "Coupling of Mechanical and Electromagnetic Fields Stimulation for Bone Tissue Engineering" (2018). Browse all Theses and Dissertations. 1950. https://corescholar.libraries.wright.edu/etd_all/1950 This Thesis is brought to you for free and open access by the Theses and Dissertations at CORE Scholar. It has been accepted for inclusion in Browse all Theses and Dissertations by an authorized administrator of CORE Scholar. For more information, please contact [email protected]. COUPLING OF MECHANICAL AND ELECTROMAGNETIC FIELDS STIMULATION FOR BONE TISSUE ENGINEERING A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Biomedical Engineering By ALYAA I. ALDEBS B.Sc. Biomedical Engineering, University of Al-Nahrain, 2012 2018 Wright State University WRIGHT STATE UNIVERSITY GRADUATE SCHOOL April 27, 2018 I HEREBY RECOMMEND THAT THE THESIS PREPARED UNDER MY SUPERVISION BY Alyaa I. Aldebs Entitled Coupling of Mechanical and Electromagnetic Fields Stimulation for Bone Tissue Engineering BE ACCEPTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Master of Science in Biomedical Engineering. ________________________________ Jaime E. Ramirez-Vick, Ph.D. Thesis Director ________________________________ Jaime E. Ramirez-Vick, Ph.D. Department Chair Committee on Final Examination ______________________________ Jaime E. Ramirez-Vick, Ph.D. ______________________________ Ulas Sunar, Ph.D. ______________________________ Nasim Nosoudi, Ph.D. _______________________________ Barry Milligan, Ph.D. Interim Dean of the Graduate School ABSTRACT ALDEBS, ALYAA I. M.S.B.M.E. Department of Biomedical, Industrial and Human Factors Engineering, Wright State University, 2018. Coupling of Mechanical and Electromagnetic Fields Stimulation for Bone Tissue Engineering. Alternative bone regeneration strategies that do not rely on harvested tissue or exogenous growth factors and cells are badly needed. However, creating living tissue constructs that are structurally, functionally and mechanically comparable to the natural bone has been a challenge so far. A major hurdle has been recreating the bone tissue microenvironment using the appropriate combination of cells, scaffold and stimulation to direct differentiation. This project presents a bone regeneration formulation that involves the use of human adipose-derived mesenchymal stems cells (hASCs) and a 3D scaffold based on a self-assembled peptide hydrogel doped with superparamagnetic nanoparticles (NPs). Osteogenic differentiation of hASCs is achieved through the direct stimulation by extremely-low frequency pulsed electromagnetic fields (pEMFs) and the indirect mechanical stimulation, through NP vibration induced by the field. This 3D construct was cultured for up to 21 days and its osteogenic capacity was assessed. Cellular morphology, proliferation, viability, as well alkaline phosphatase activity, calcium deposition were monitored during this time. The results show that the pEMFs have no negative effect on cell viability and induce early differentiation of hASCs to an osteoblastic phenotype when compared to a iii cell without biophysical stimulation. This effect results from the synergy between the pEMF and NP that acts as remote stimulation of the mechanotransduction pathways which activate biochemical signals between cells to go under differentiation or proliferation. The use of this approach offers a safe and effective treatment option for the treatment of non-union bone fractures. In addition, this formulation can be directly injected into the wound site, making it minimally invasive as well. iv Table of Contents ABSTRACT ...................................................................................................................... iii Table of Contents .............................................................................................................. v List of Figures .................................................................................................................. vii Acknowledgment .............................................................................................................. ix CHAPTER I ...................................................................................................................... 1 Introduction ....................................................................................................................... 1 1.1 Project overview ........................................................................................................ 1 1.2 Objectives .................................................................................................................. 5 CHAPTER II ..................................................................................................................... 7 Literature Review ............................................................................................................. 7 2.1 Bone structure ........................................................................................................... 7 2.2 Bone anatomy ............................................................................................................ 7 2.3 Bone composition .................................................................................................... 11 2.3.1 Bone Extracellular Matrix ................................................................................ 11 2.3.2 bone cells .......................................................................................................... 14 2.3.3 growth factors ................................................................................................... 17 2.4 Bone diseases .......................................................................................................... 19 2.5 Bone replacement and tissue engineering ............................................................... 20 2.6 Cellular sources for bone tissue engineering........................................................... 22 2.7 Three-dimensional scaffold for bone tissue engineering..……..………………25 2.7.1 Self-assembled peptide scaffold ....................................................................... 26 2.8 Biophysical stimulation ........................................................................................... 30 2.8.1 Effect of mechanical stimulation ...................................................................... 30 2.8.2 Effect of electromagnetic fields ........................................................................ 34 v 2.9 Effect of magnetic scaffold under EMF stimulation for bone regeneration ........... 38 CHAPTER III ................................................................................................................. 46 Materials and Methods ................................................................................................... 46 3.1 Cell culture .............................................................................................................. 46 3.2 Three-dimensional cells encapsulation and gel formation .............................. 46 3.3 Cells viability assay ................................................................................................. 47 3.4 Cells differentiation assay ....................................................................................... 48 3.5 Measurement of mineralization ............................................................................... 49 3.6 Cells morphology .................................................................................................... 49 3.7 Alkaline phosphatase staining ................................................................................. 50 3.8 Fourier transform infrared spectroscopy (FTIR) analysis ....................................... 51 3.9 Electromagnetic fields exposure system ................................................................. 51 3.10 Statistical analysis ................................................................................................. 53 CHAPTER IV.................................................................................................................. 54 Results &Discussion ........................................................................................................ 54 4.1 Results & Discussion ..................................................................................... 54 4.1.1 Cell viability ............................................................................................ 54 4.1.2 Differentiation to osteoblasts ................................................................... 59 4.1.3 Mineralization .......................................................................................... 63 4.1.4 Cell morphology ...................................................................................... 65 4.1.5 Alkaline phosphatase staining .................................................................. 68 4.1.6 FTIR analysis ..........................................................................................