Synthesis of functionalized-thermo responsive-water soluble co-polymer for conjugation to protein for biomedical applica
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Synthesis of functionalized-thermo responsive-water soluble co-polymer for conjugation to protein for biomedical applications Keywords: Protein, synthetic polymer, bioconjugation, hydrogel, tissue engineering, biofactor delivery, Ali Fathi1, Hua Wei1, Wojciech Chrzanowski2, Anthony S. Weiss3, Fariba Dehghani1
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School of Chemical and Biomolecular Engineering, the University of Sydney, NSW 2006, Australia Faculty of Pharmacy, the University of Sydney, NSW 2006, Australia, School of Molecular Bioscience, the University of Sydney, NSW 2006, Australia,
ABSTRACT The aim of this study was to develop a thermo-responsive and bioactive polymer with suitable mechanical properties for musculoskeletal tissue engineering applications. A copolymer was synthesized that comprised of hydrophilic polyethylene glycol, thermo responsive Nisopropylacrylamide (NIPAAm), 2-hydroxyethyl methacrylate-poly(lactide) (HEMA-PLA) to enhance mechanical strength and an active N-acryloxysuccinimide (NAS) group for conjugation to proteins to enhance biological properties. A model protein such as elastin was used to assess the feasibility of conjugating this polymer to protein. The results of 1HNMR analyses confirmed that random polymerization was viable technique for synthesis of this copolymer. The copolymers synthesized with PEG content of 3 mol% were water soluble. A hydrogel was created by dissolving the copolymer and elastin below room temperature in aqueous media, followed by rapid gelation at 37°C. The results of Fourier transform infrared analyses confirmed the conjugation of protein to copolymer due to significant reduction of ester group absorption (1735 cm-1). This data confirmed molecular interaction between protein and the temperature responsive co-polymer. Our preliminary results demonstrated that it is viable to tune different properties of this hydrogel by changing the composition of co-polymer. INTRODUCTION Injectable hydrogels are promising as a minimally invasive approach for tissue engineering [1]. The immense interest to injectable hydrogels is due to their tunable physical and mechanical properties [2–4], and their high water intake capacity [5]. In this approach, the polymer solution is loaded with targeted cells, and injected in vivo. It thereafter undergoes solution-to-gel transition (sol-gel) in situ due to different chemical or physical stimuli [6–8]. Thermogelation process is an ideal and benign scheme for in situ gelation of polymers. In this case a hydrogel is formed when the temperature is shifted to above the lower critical solution temperature (LCST) of thermo-responsive polymer. [9]. Copolymerization of different stimuli-responsive naturally derived polymers such as elastin like polypeptides (ELP) [10–12], gelatin, chitosan [13], hyaluronic acid derived copolymers [14], and synthetic polymers such as poly(Nisopropylacrylamide) (pNIPAAm) [15] has been deemed to be an effective approach to develop polymers with tunable physiochemical, gelation and biochemical properties. PNIPAAm based copolymers are widely used
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