Dynamics of smooth muscle cell deadhesion from thermosensitive hydroxybutyl chitosan

Abstract

Thermoresponsive polymer (TRP) enables the enzyme-free harvesting of cells through an acute increase in surface hydrophilicity of TRP across its lower critical solution temperature (LCST), rendering feasible the generation of polymer-free cell sheets for regenerative medicine applications. To date, the intricate mechanisms of cell deadhesion/detachment on TRP surface remain obscure. Elucidation of such biophysical responses would be valuable for the cell sheet technology. In this study, integrative biophysical techniques are applied to probe the thermal-induced deadhesion kinetics of smooth muscle cell (SMC) on thermoresponsive hydroxybutyl chitosan (HBC29) against different periods of pre-culture time at 37 °C. Atomic force microscopy demonstrates that both the surface topography and mechanical property of HBC29 film in water are acutely modulated across its LCST. Firstly, cells show negligible changes in adhesion contact area during low-temperature incubation on unmodified tissue culture polystyrene (TCPS). Secondly, the recession of adhesion contact and retraction of cell body for cells with different pre-culture times are triggered by HBC29 coating on TCPS. Interestingly, the initial rate of reduction in the normalized adhesion contact area of SMC is negatively correlated with the pre-culture time. Thirdly, the degree of cell deformation and average adhesion energy are reducing functions of time only for SMCs with the lowest preculture time. In contrast, adhesion energy per cell is a reducing function of time irrespective of the change of pre-culture time. Lastly, the temporal dynamics of cytoskeleton organization and β-actin/ smoothelin-B mRNA expression for SMCs is strongly dependent on the pre-culture time. Overall, this study demonstrates that the thermal-induced deadhesion of SMC on TRP is characterized by the evolution of its contractile phenotypes.

The use of biodegradable polymeric nanoparticles in combination with a low-pressure gene gun for transdermal DNA delivery

“…CS-based nanoparticles, by their reduced cytotoxicity and ability to transport and release genes intracellularly, where expression of the encoded protein occurs, are considered as a potential non-viral delivery system for plasmid DNA [17]. In this study, the feasibility of using CS-based nanoparticles with or without the incorporation of g-PGA as a carrier for transdermal DNA delivery using a low-pressure gene gun was evaluated.…”

Microarrays of over 2000 hydrogels – Identification of substrates for cellular trapping and thermally triggered release

“…Thermosensitive gels/polymers have, for example, been applied in tissue engineering [3][4][5], gene delivery [6,7], reversible molecule adsorption [8], modulation of protein-ligand binding [9], 'smart' actuators [10,11] and liquid micro-lenses [12], although perhaps the most wide spread interest is as a surface coating for cellular binding, which allows cell harvesting without recourse to traditional methods (scratching or proteases) which can damage the cells or cleave surface proteins [13][14][15].…”

Preparation and adsorption kinetics of porous γ-glycidoxypropyltrimethoxysilane crosslinked chitosan–β-cyclodextrin membranes

“…exploited in different areas in food science, biochemistry, pharmaceuticals, medicine, agriculture [9][10][11][12][13][14].…”