Incorporated Copolymers and Blends for Bone Regeneration
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University of Kentucky UKnowledge Theses and Dissertations--Biomedical Engineering Biomedical Engineering 2017 SYNTHESIS AND CHARACTERIZATION OF POLY(SIMVASTATIN) - INCORPORATED COPOLYMERS AND BLENDS FOR BONE REGENERATION Theodora Asafo-Adjei University of Kentucky, [email protected] Author ORCID Identifier: https://orcid.org/0000-0001-8124-9040 Digital Object Identifier: https://doi.org/10.13023/ETD.2017.362 Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Asafo-Adjei, Theodora, "SYNTHESIS AND CHARACTERIZATION OF POLY(SIMVASTATIN) - INCORPORATED COPOLYMERS AND BLENDS FOR BONE REGENERATION" (2017). Theses and Dissertations--Biomedical Engineering. 46. https://uknowledge.uky.edu/cbme_etds/46 This Doctoral Dissertation is brought to you for free and open access by the Biomedical Engineering at UKnowledge. 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Theodora Asafo-Adjei, Student Dr. David Puleo, Major Professor Dr. Abhijit Patwardhan, Director of Graduate Studies SYNTHESIS AND CHARACTERIZATION OF POLY(SIMVASTATIN) - INCORPORATED COPOLYMERS AND BLENDS FOR BONE REGENERATION _____________________________ DISSERTATION _____________________________ A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the College of Engineering at the University of Kentucky By Theodora Atta Asafo-Adjei Lexington, Kentucky Advisor: Dr. David Puleo, Professor of Biomedical Engineering Co-Advisor: Dr. Thomas Dziubla, Professor of Chemical and Materials Engineering Lexington, Kentucky Copyright © Theodora Atta Asafo-Adjei 2017 ABSTRACT OF DISSERTATION SYNTHESIS AND CHARACTERIZATION OF POLY(SIMVASTATIN) - INCORPORATED COPOLYMERS AND BLENDS FOR BONE REGENERATION Common biodegradable polyesters such as poly(lactic acid) (PLA), poly(lactic- co-glycolic acid) (PLGA) and poly(ε-caprolactone) (PCL) are used as drug delivery vehicles for tissue regenerative applications. However, they are typically bioinert, with drug loading limitations. Polymerizing the active agent or precursor into its respective biodegradable polymer would control drug loading via molar ratios of drug to initiator used for synthesis. Simvastatin was chosen due to its favorable anti-inflammatory, angiogenic, and osteogenic properties. In addition, its lactone ring lends itself to ring- opening polymerization and, consequently, the synthesis of poly(simvastatin) with controlled simvastatin release. Simvastatin was first polymerized with a 5kDa methyl-terminated poly(ethylene glycol) (mPEG) initiator and catalyzed via stannous octoate to form poly(simvastatin)- block-poly(ethylene glycol). Molecular weights ranged from 9.5kDa, with a polydispersity index (PDI) of 1.1 at 150 °C, to 75kDa with a PDI of 6.9 at 250 °C. First- order propagation rates were seen. Infrared spectroscopy showed carboxylic and methyl ether stretches unique to simvastatin and mPEG in the copolymer, respectively. Slow degradation was seen in neutral and alkaline conditions, with simvastatin, simvastatin- incorporated macromolecules, and mPEG identified as degradation products. Alternatively, triazabicyclodecene (TBD) was used to mediate simvastatin polymerization. A lower temperature of 150°C led to successful polymerization using 5kDa mPEG, compared to at least 200 °C via stannous octoate. TBD was also successful for reactions using 2 or 0.55kDa mPEG. The biodegradability of poly(simvastatin)- block-poly(ethylene glycol) via TBD improved, losing twice more mass in phosphate- buffered saline, pH 7.4, than the copolymer synthesized via stannous octoate. Release rates of three different copolymers synthesized demonstrated tunable simvastatin release. To further modulate degradation, poly(simvastatin)-block-poly(ethylene glycol) was blended with 5, 2, or 0.55kDa mPEG-initiated PLA copolymers. The blends showed a compressive elastic modulus ranging from 26 to 44MPa, within the magnitude of trabecular bone (approximately 50MPa). Tunability in mass loss and release was also seen due to varied ratios of incorporated PLA copolymers. Lastly, copolymer degradation byproducts inhibited HMG-CoA reductase and showed possible enhancement of osteoblastic activity in vitro. A pilot study using a rodent calvarial onlay model showed tolerability of the polymers and potential for long- term evaluations of bioactivity. Poly(simvastatin) may be useful in regenerative applications. Keywords: poly(simvastatin), ring-opening polymerization, drug delivery, simvastatin, regenerative applications Theodora Atta Asafo-Adjei Student Signature 7/6/2017 Date SYNTHESIS AND CHARACTERIZATION OF POLY(SIMVASTATIN) - INCORPORATED COPOLYMERS AND BLENDS FOR BONE REGENERATION By Theodora Atta Asafo-Adjei Dr. David Puleo Advisor of Dissertation Dr. Thomas Dziubla Co-Advisor of Dissertation Dr. Abhijit Patwardhan Director of Graduate Studies 7/6/2017 Date ACKNOWLEDGEMENTS The number of friends and colleagues who have helped me push through during my time at UK are endless. I would like to thank my former labmates Matt Brown, Sharath Sundararaj, Amanda Rudd, Paul Fisher, Bryan Orellana, Sandeep Ramineni, Cheryl Jennings, Rohith Jaraham, Yuan Zou, Amir Najarzadeh, Michelle Fuentes, Matt Rudd, Nicholas Anderson, Nicholas Kohrs, Alexander Chen, Andrew Phan, Nandakumar Venkatesan, Thilanga Liyanage, Paritosh Wattamwar, David Cochran, Sundar Authimoolam, Prachi Gupta, Vinod Patil, Andrew Lakes, and Carolyn Jordan. There are many others, but always having you guys around to bounce ideas off of and help out in and out of lab was such a lifesaver for me. I would like to thank my advisors Dr. Puleo, for your understanding in allowing me to find my voice while guiding me to ask the most relevant questions in designing experiments, and Dr. Dziubla, for pushing me to think past the question in front of me while providing insightful perspectives on ways to investigate them. You two provided the ideal balance in our meetings to realize what work it would take to answer the question at hand, and resolve experimental setbacks while providing the motivation needed to navigate through the project. To my close friends, thank you for always checking up on me, being great cooks, giving me sound advice about life, and just being the occasional escape I needed to get away from the many obstacles I had in lab. Last but not least, I’d like to thank my family. Thanks to my younger siblings, iii Olivia and Miles, for being my rocks of support through these years, my dad who taught me that goals are always attainable with hard work, and mom who has instilled in me strength and resilience taught only by the actions of her own. I am so grateful for how involved you all have been throughout my journey here. I’ve learned so much, as a result. Know that you will always be appreciated. It was a challenging, but fun experience! iv Table of Contents ACKNOWLEDGMENTS..................................................................................................iii List of Tables ...................................................................................................................viii List of Figures.....................................................................................................................ix Ch. 1 Introduction…………………………………………………...………………..….1 Ch. 2 Background and Significance………………………...…………………………..5 2.1 Bone Grafts and Substitutes…………………………………………………………...5 2.1.1 Autologous and Allogenic Bone Grafts……………………………………………..5 2.1.2 Calcium-Based Substitutes ……………………………………………………..…..6 2.2 Common Biodegradable Polymers……………………………………………..……..7 2.2.1 Poly(lactic acid) (PLA)……………………………………………………………...7 2.2.2 Poly(lactic-co-glycolic acid) (PLGA)…………………………………………...…..8 2.2.3 Poly(ε-caprolactone) (PCL)…………………………………………………………9 2.2.4 Poly(anhydrides)…………………………………………………………………...10 2.3 Polymer Synthesis……………………………………………………………………12