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Development of Drug-Eluting Surgical Sutures for the Wound- Healing Process Using Melt Extrusion Technology

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Development of Drug-Eluting Surgical Sutures for the Wound- Healing Process Using Melt Extrusion Technology Development of Drug-Eluting Surgical Sutures for the Wound- Healing Process Using Melt Extrusion Technology Xiaoxuan Deng A thesis submitted for the degree of Master of Science at the University of Otago, Dunedin, New Zealand. February 2020 ABSTRACT Incorporation of an anti-inflammatory drug with polymer materials to produce suture products is important especially when the suture is used for internal organs and/or tissues. In this work, poly(ε-caprolactone) (PCL), polyethylene glycol 400 (PEG400), chitosan and keratin were used as biopolymer matrix to encapsulate 5, 15, 30 wt% diclofenac potassium (DP) to achieve controlled drug release. At first, polymer alone was extruded by hot- melt extrusion technique under various temperatures and at different PCL/PEG400/chitosan-keratin ratios. The products were analysed in terms of FTIR characterization, thermal properties and mechanical properties. According to the characterization analysis of polymer sutures, the optimal polymer formulation (PCL: PEG400: chitosan/keratin= 80:19:1 wt%) (Group 1) was chosen to manufacture sutures with drug DP content at 5, 15, 30 wt%. In addition, PCL/PEG400/chitosan (Group 2) and PCL/PEG400/keratin (Group 3) were also fabricated with drug DP to create sutures, the polymer and drug weight ratios were 95:5, 85:15 and 70:30 wt%. Drug release features were investigated via the drug dissolution test. MTT assay, live/dead assay and scratch assay were also applied to study the biocompatibility of drug embedded sutures. Results showed that sutures can be extruded with a smooth surface and uniform thickness under temperature of 63 ± 1 °C. It was observed from DSC and TGA studies that completely amorphous and miscible solid dispersions were prepared. FTIR analysis indicated that the presence of hydrogen bonds between the polymers improved material miscibility. Tensile properties of the sutures were clearly affected by PEG400, chitosan and keratin addition. The optimal formulation of tensile strength was obtained when PEG400: chitosan/keratin was 19:1 wt%. Rapid drug release rate was observed at different stages of group 1, 2, 3. For group 1, the release occurred rapidly occurred after 1 h 45 mins, whereas group 2 exhibited fast release between 1 h 45 mins and 3 h 45 mins. Group 3 displayed a sustained release where a fast release occurred from the onset to 3 h 45 mins. However, the final concentration of drug release in group 1 was lower than the other two groups, which can be predicted that a longer drug release time would be in an extended period compared to the other two groups. According to biocompatibility test, group 1 demonstrated very high cell viability at 72 h. Especially ii sample F3, which contained 30 wt% drug, and the ratio of PCL/PEG/chitosan-keratin was 80/19/1 w/w. It achieved the highest wound healing rate in the scratch assay. iii ACKNOWLEDGEMENTS I would first like to thank my supervisor Associate Prof. Azam Ali of the Centre for Bioengineering and Nanomedicine, Department of Food Science. He has been my supervisor since the first day I came to University of Otago to start my postgraduate study. His advice and expertise were priceless to me and he was willing to answer my questions and doubts at any of his free time. Owing to his help and suggestions, I could work on this interesting research topic, and will pursue knowledge on this path further. He encouraged me in my work and guided me on the right track when I was confused. We had fortnightly meeting on Thursday, which was really helpful. I would also like to thank Dr. Maree Gould of the Centre for Bioengineering and Nanomedicine, Department of Food Science, who was never tired of explaining virous cell culture concepts and analysis technique. Helping me familiar with operating procedure for MTT assay, live/dead assay and scratch assay, and recommending me with Prism software which made the graphic building and statistical analysing much easier. I could not complete my data analysis in a relatively short time without her great help. On the other hand, she is also a friendly and funny person, who always treats people with kindness and lots of harmless jokes. I never lose smile on my face whenever talking to her. I will forever be grateful to Ian Stewart of the Department of Chemistry whose immense help and guidance allowed me to carry out TGA and DSC experiments. Many thanks also to fellow PhD students Tajul Islam for his patient explaining whenever I felt confused about experimental steps or concepts, and Stephen Giteru for his training for mechanical testing. I would like to thank my friend Bhavini Patel, who invited me to go to gym together and shared life experience with me, I often felt inspired just by chatting with her. I must also express my infinite gratitude to Dino Milotic who sacrificed lots of time reading and editing my thesis, giving me detailed feedback. He also gave me suggestion and encouraged me when I needed it the most. Finally, I have to thank my parents who provided me this great opportunity to come to University of Otago studying Master of iv Science. This accomplishment would not have been happened without their encouragement and financial support. v TABLE OF CONTENTS Abstract .......................................................................................................................... ii Acknowledgements ....................................................................................................... iv Table of Contents .......................................................................................................... vi List of Tables ................................................................................................................ ix List of Figures ................................................................................................................ x List of Abbreviations ................................................................................................... xii 1 Introduction ................................................................................................................. 1 1.1 Antimicrobial Sutures ...................................................................................................... 1 1.2 Classification of Suture Material ..................................................................................... 4 1.2.1 Absorbable Sutures ................................................................................................................. 6 1.2.2 Non-Absorbable Sutures ......................................................................................................... 9 1.3 Sutures with Active Pharmaceutical Ingredients (APIs) ............................................... 11 1.4 Suture Structure ............................................................................................................. 14 1.4.1 Monofilament and Multifilament (Braided) Sutures ............................................................. 14 1.4.2 Barbed Sutures ...................................................................................................................... 15 1.4.3 Smart Sutures ........................................................................................................................ 16 1.5 Fabrication Process ........................................................................................................ 16 1.5.1 Electrospinning ..................................................................................................................... 24 1.5.2 Melt Extrusion ...................................................................................................................... 26 1.5.3 Coating .................................................................................................................................. 28 1.6 Conclusion ..................................................................................................................... 29 2 The Current Project ................................................................................................... 31 2.1 Fabrication of Drug-Eluting Sutures with Hot- melt Extrusion Technique................... 31 2.2 Materials Selected for This Study ................................................................................. 32 2.3 Aims and Objectives...................................................................................................... 33 3 Materials and Methods .............................................................................................. 35 3.1 Materials ........................................................................................................................ 35 3.2 Equipment ..................................................................................................................... 35 3.3 Suture Preparation for Control Group ........................................................................... 36 3.4 Fourier Transformation-Infrared Spectroscopy (FT-IR) ............................................... 39 3.5 Thermal Gravimetric Analysis (TGA) .......................................................................... 40 3.6 Differential Scanning Calorimetry Analysis (DSC) ...................................................... 41 vi 3.7 Mechanical Properties ................................................................................................... 42 3.8 Suture Preparation for Formulation Group .................................................................... 43 3.9 Drug
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