Engineered chitosan for improved 3D tissue growth through Paxillin-FAK-ERK activation

Kafi, M. A., Aktar, M. K., Todo, M. and Dahiya, R. (2020) Engineered chitosan for improved 3D tissue growth through Paxillin-FAK-ERK activation. Regenerative Biomaterials, 7(2), pp. 141-151. (doi: 10.1093/rb/rbz034) (PMID:32296533) (PMCID:PMC7147363)

Abstract

Scaffold engineering has attracted significant attention for three-dimensional (3D) growth, proliferation and differentiation of stem cells in vitro. Currently available scaffolds suffer from issues such as poor ability for cell adhesion, migration and proliferation. This paper addresses these issues with 3D porous chitosan scaffold, fabricated and functionalized with cysteine-terminated Arg-Gly-Asp (Cys-RGD) tri-peptide on their walls. The study reveals that the compressive moduli of the scaffold is independent to RGD functionalization but shows dependence on the applied freezing temperature (TM) during the fabrication process. The low freezing TM (−80°C) produces scaffold with high compressive moduli (14.64 ± 1.38 kPa) and high TM (−30°C) produces scaffold with low compressive moduli (5.6 ± 0.38 kPa). The Cys-RGD functionalized scaffolds lead to significant improvements in adhesion (150%) and proliferation (300%) of human mesenchymal stem cell (hMSC). The RGD-integrin coupling activates the focal adhesion signaling (Paxillin-FAK-ERK) pathways, as confirmed by the expression of p-Paxillin, p-FAK and p-ERK protein, and results in the observed improvement of cell adhesion and proliferation. The proliferation of hMSC on RGD functionalized surface was evaluated with scanning electron microscopy imaging and distribution though pore was confirmed by histochemistry of transversely sectioned scaffold. The hMSC adhesion and proliferation in scaffold with high compressive moduli showed a constant enhancement (with a slope value 9.97) of compressive strength throughout the experimental period of 28 days. The improved cell adhesion and proliferation with RGD functionalized chitosan scaffold, together with their mechanical stability, will enable new interesting avenues for 3D cell growth and differentiation in numerous applications including regenerative tissue implants.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Kafi, Md Abdul and Dahiya, Professor Ravinder
• Authors: Kafi, M. A., Aktar, M. K., Todo, M., and Dahiya, R.
• College/School: College of Science and Engineering > School of Engineering > Electronics and Nanoscale Engineering
• Journal Name: Regenerative Biomaterials
• Publisher: Oxford University Press
• ISSN: 2056-3418
• ISSN (Online): 2056-3426
• Published Online: 30 September 2019
• Copyright Holders: Copyright © 2019 The Authors
• First Published: First published in Regenerative Biomaterials 7(2): 141-151
• Publisher Policy: Reproduced under a Creative Commons License

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
71876	1	BEND	Ravinder Dahiya	European Commission (EC)	704807	ENG - ENGINEERING ELECTRONICS & NANO ENG
66386	1	Engineering Fellowships for Growth: Printed Tactile SKIN	Ravinder Dahiya	Engineering and Physical Sciences Research Council (EPSRC)	EP/M002527/1	ENG - ENGINEERING ELECTRONICS & NANO ENG