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

Md Abdul Kafi; Khudishta Aktar; Mitsugu Todo; Ravinder Dahiya

doi:10.1093/rb/rbz034

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

Regen Biomater. 2020 Mar;7(2):141-151. doi: 10.1093/rb/rbz034. Epub 2019 Sep 30.

Authors

Md Abdul Kafi^{1

2

3}, Khudishta Aktar¹, Mitsugu Todo³, Ravinder Dahiya¹

Affiliations

¹ BEST Group, School of Engineering, University of Glasgow, Glasgow G12 8QQ, UK.
² Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.
³ Research Institute for Applied Mechanics, Kyushu University, Kasuga, Fukuoka, Japan.

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.

Keywords: Cys-RGD; adhesion signaling; chitosan; hMSC; porous scaffold; proliferation; tissue engineering.