Department of Food Processing Technology and Management

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    THE INFLUENCE OF SWELLING ON LOCAL ELASTIC PROPERTIES OF POLYACRYLAMIDE HYDROGELS
    (Open Science Framework, 2017) Ramesh, Subramani; Alicia, Izquierdo-Alvarez; Pinaki, Bhattacharya; Mathieu, Meerts; Paula, Moldenaers; Herman, Ramon; Hans Van, Oosterwyck
    Polyacrylamide (PAM)hydrogelsarecommonly usedas soft cell culture substrates for cell mechanical and mechanobiological studiesbecause oftheir tunable stiffness,easeof handling, transparent natureand surface functionalization to promote cell adhesion. The dependence of bulk rheological and local elastic properties (for example, as assessed by means of Atomic Force Microscopyor AFM) of PAM hydrogels onmonomer and cross-linkerconcentrationsand on polymerization temperature havebeen extensively investigated.PAM hydrogels are known to swell, which may affect their elastic properties and therefore may complicate the interpretation ofcell culturingexperiments because of a lack of control of substrate stiffness. Direct measurements of the effect of swelling on PAM elastic properties are scarce. We report here, for the first time, the direct observation of swelling (by measuring the volumetric swelling ratio) and its influence onlocal elastic properties, as measuredby AFM. Bulk rheological measurements were performed as well to enable the comparison between local and global elastic properties during and after hydrogel polymerization. Four PAM hydrogel compositions were considered,with corresponding storage shear moduliof 4530 Pa(termed stiffest), 2900 Pa(stiff), 538 Pa(soft)and 260 Pa(softest) as measured immediately after polymerization. These values as well as values obtained during hydrogel polymerization were found to be in good agreement with the local elastic moduli derived from AFM and assuming hydrogel incompressibility. After polymerization, the hydrogels were subjected to swelling conditions over six days in phosphate buffered salineand swelling ratios and local elastic moduli were measured each day.Additional short term measurements (at 0, 3, 6, 9, 12 and 24 hours) were performed for the soft and softest gelsto study their swelling kinetics in more detail. Swelling ratio and local elastic modulus did not change with time for the stiffest and stiff gels, while for the soft and softest gels substantialchanges between Day 0 and Day 1were found for both swelling ratio (21.6%and 133% increase for soft and softest gel respectively) and local elastic modulus (33.7%and 33.3% decrease for soft and softest gel respectively). Experimental data werefurther analysed theoretically by combining models of ideal elastomeric gels with a poroelastic swelling kinetics model, which confirmed the validity of observed trends with respect to literature data. The results demonstrate that swelling can have an important effect on PAM elastic properties and must be taken into account when using PAM as a cell culture substrate, particularly for PAM gels with low monomer and cross-linker concentrations
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    THE INFLUENCE OF SWELLING ON ELASTIC PROPERTIES OF POLYACRYLAMIDE HYDROGELS
    (Frontiers in Materials, 2020-07-23) Ramesh, Subramani; Alicia, Izquierdo-Alvarez; Pinaki, Bhattacharya; Mathieu, Meerts; Paula, Moldenaers; Herman, Ramon; Hans, Van Oosterwyck
    Polyacrylamide (PAM) hydrogels are commonly used as substrates for cell mechanical and mechanobiological studies because of their tunable stiffness and ease of handling. The dependence of bulk rheological and local elastic properties (assessed by Atomic Force Microscopy, or AFM) of PAM hydrogels on its composition and polymerization temperature has been extensively studied. PAM hydrogels swell when immersed in media, but the influence of swelling on local elastic properties is poorly characterized. Direct measurements of the effect of swelling on PAM elastic properties are scarce. We report here, for the first time, the direct measurements of volumetric swelling and local elastic properties of PAM gels throughout the post-polymerization swelling process until equilibrium. First, local and global elastic properties (measured by rheology), were obtained during polymerization in the absence of swelling, and showed good agreement with each other. Four PAM hydrogel compositions were characterized thus, with corresponding storage shear moduli (as measured immediately after polymerization) of 4,530 Pa (termed stiffest), 2,900 Pa (stiff), 538 Pa (soft), and 260 Pa (softest). Next, all compositions were subjected to swelling in phosphate buffered saline. Swelling ratios and local elastic moduli were measured at 0, 3, 6, 9, 12, and 24 h post-polymerization for the soft and softest compositions, and once daily till 6 days post-polymerization for all four compositions. For the stiffest and stiff gels, swelling ratio, and local elastic modulus changed negligibly with time, while for the soft and softest gels, substantial changes between Day 0 and Day 1 were found for both swelling ratio (increased by 21.6 and 133%, respectively), and local elastic modulus decreased (by 33.7 and 33.3%, respectively), substantially. Experimental data were analyzed by a model that combined ideal elastomer mechanics and poroelastic swelling kinetics model. Model predictions confirmed the validity of present measurements with respect to past studies where swelling and elastic properties were not measured simultaneously. The present study underlines the important effect swelling can have on PAM elastic properties and provides detailed quantitative data to guide the duration taken to reach equilibrium—a useful information for cell mechanics experiments. In addition, the simultaneous measurements of swelling and local elastic moduli provide novel data for the validation of theoretical models.