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Biocompatibility evaluation of protein-incorporated electrospun polyurethane-based scaffolds with smooth muscle cells for vascular tissue engineering
Authors:Lin Jia  Molamma P. Prabhakaran  Xiaohong Qin  Dan Kai  Seeram Ramakrishna
Affiliation:1. Key Laboratory of Textile Science & Technology, Ministry of Education, College of Textiles, Donghua University, No. 2999 North Renmin Road, Songjiang, Shanghai, 201620, China
2. Faculty of Engineering, Center for Nanofibers and Nanotechnology, E3-05-14, Nanoscience and Nanotechnology Initiative, National University of Singapore, 2 Engineering Drive 3, Singapore, 117576, Singapore
3. NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore, Singapore
Abstract:Nanotechnology has enabled the engineering of a variety of materials to meet the current challenges and needs in vascular tissue regeneration. In this study, four different kinds of native proteins namely collagen, gelatin, fibrinogen, and bovine serum albumin were incorporated with polyurethane (PU) and electropsun to obtain composite PU/protein nanofibers. SEM studies showed that the fiber diameters of PU/protein scaffolds ranged from 245 to 273 nm, mimicking the nanoscale dimensions of native ECM. Human aortic smooth muscle cells (SMCs) were cultured on the electrospun nanofibers, and the ability of the cells to proliferate on different scaffolds was evaluated via a cell proliferation assay. Cell proliferation on PU/Coll nanofibers was found the highest compared to other electrospun scaffolds and it was 42 % higher than the proliferation on PU/Fib nanofibers after 12 days of cell culture. The cell–biomaterial interaction studies by SEM confirmed that SMCs adhered to PU/Coll and PU/Gel nanofibers, with high cell substrate coverage, and both the scaffolds promoted cell alignment. The functionality of the cells was further demonstrated by immunocytochemical analysis, where the SMCs on PU/Coll and PU/Gel nanofibers expressed higher density of SMC proteins such as alpha smooth muscle actin and smooth muscle myosin heavy chain. Cells expressed biological markers of SMCs including aligned spindle-like morphology on both PU/Coll and PU/Gel with actin filament organizations, better than PU/Fib and PU/BSA scaffolds. Our studies demonstrate the potential of randomly oriented elastomeric composite scaffolds for engineering of vascular tissues causing cell alignment.
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