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Click chemistry approach for fabricating PVA/gelatin nanofibers for the differentiation of ADSCs to keratinocytes
Authors:Rajeswari Ravichandran  Jayarama Reddy Venugopal  Subramanian Sundarrajan  Shayanti Mukherjee  John Forsythe  Seeram Ramakrishna
Affiliation:1. Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, Block E3, #05-12, 2 Engineering Drive 3, Singapore, 117576, Singapore
2. Department of Mechanical Engineering, National University of Singapore, Singapore, 117576, Singapore
3. Department of Materials Engineering, Monash University, Melbourne, VIC, Australia
Abstract:Every year, millions of people suffer from dermal wounds caused by heat, fire, chemicals, electricity, ultraviolet radiation or disease. Tissue engineering and nanotechnology have enabled the engineering of nanostructured materials to meet the current challenges in skin treatments owing to such rising occurrences of accidental damages, skin diseases and defects. The abundance and accessibility of adipose derived stem cells (ADSCs) may prove to be novel cell therapeutics for skin regeneration. The nanofibrous PVA/gelatin/azide scaffolds were then fabricated by electrospinning using water as solvent and allowed to undergo click reaction. The scaffolds were characterized by SEM, contact angle and FTIR. The cell–scaffold interactions were analyzed by cell proliferation and the results observed that the rate of cell proliferation was significantly increased (P ≤ 0.05) on PVA/gelatin/azide scaffolds compared to PVA/gelatin nanofibers. In the present study, manipulating the biochemical cues by the addition of an induction medium, in combination with environmental and physical factors of the culture substrate by functionalizing with click moieties, we were able to drive ADSCs into epidermal lineage with the development of epidermis-like structures, was further confirmed by the expression of early and intermediate epidermal differentiation markers like keratin and filaggrin. This study not only provides an insight into the design of a site-specific niche-like microenvironment for stem cell lineage commitment, but also sheds light on the therapeutic application of an alternative cell source—ADSCs, for wound healing and skin tissue reconstitution.
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