Neuronal Differentiation Modulated by Polymeric Membrane Properties

Abstract

In this study, different collagen-blend membranes were successfully constructed by blending collagen with chitosan (CHT) or poly(lactic-co-glycolic acid) (PLGA) to enhance their properties and thus create new biofunctional materials with great potential use for neuronal tissue engineering and regeneration. Collagen blending strongly affected membrane properties in the following ways: (i) it improved the surface hydrophilicity of both pure CHT and PLGA membranes, (ii) it reduced the stiffness of CHT membranes, but (iii) it did not modify the good mechanical properties of PLGA membranes. Then, we investigated the effect of the different collagen concentrations on the neuronal behavior of the membranes developed. Morphological observations, immunocytochemistry, and morphometric measures demonstrated that the membranes developed, especially CHT/Col30, PLGA, and PLGA/Col1, provided suitable microenvironments for neuronal growth owing to their enhanced properties. The most consistent neuronal differentiation was obtained in neurons cultured on PLGA-based membranes, where a well-developed neuronal network was achieved due to their improved mechanical properties. Our findings suggest that tensile strength and elongation at break are key material parameters that have potential influence on both axonal elongation and neuronal structure and organization, which are of fundamental importance for the maintenance of efficient neuronal growth. Hence, our study has provided new insights regarding the effects of membrane mechanical properties on neuronal behavior, and thus it may help to design and improve novel instructive biomaterials for neuronal tissue engineering.

Environmental Elasticity Regulates Cell-type Specific RHOA Signaling and Neuritogenesis of Human Neurons

“…To further investigate the effect of environmental elasticity on neurite development, we examined neurite lengths in 3D collagen. Numerous studies have used collagen I matrices to study the effects of variable elastic conditions on cell shape, polarity, and durotaxis (Sundararaghavan et al., 2009, Willits and Skornia, 2004), as well as differentiation, neuritogenesis, and nerve regeneration (Gil and del Rio, 2012, Morelli et al., 2017, Musah et al., 2014). To test whether altering collagen elasticity affected hMN and hFB neurite outgrowth in three dimensions, we cultured neurospheres on top of 1.5 mg/mL fibrillar collagen I.…”

PLGA Multiplex Membrane Platform for Disease Modelling and Testing of Therapeutic Compounds

“…Besides stiffness, ultimate tensile strength (UTS) and elongation at break [ε] also influence the overall biomechanical characteristics of the PLGA membrane, which have been defined and reported in Table 1. In a previous work, aimed at disclosing the effect of membrane mechanical properties for the reconstruction of a neuronal tissue analogue [15], we showed that UTS and elongation at break are two key parameters able to influence growth cone formation and axonal and dendritic complex arborization. The mechanical features of brain tissue are quite peculiar, being anisotropic and extremely soft (e.g., <1 kPa) [22].…”

“…Membranes of Poly(D,L-lactide-co-glycolide) (PLGA) were prepared in a flat configuration using a phase inversion technique with solvent evaporation, as described elsewhere [15]. In brief, PLGA (10% w/v) (MW 50,000-75,000 Da, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in 1.4-dioxan.…”

“…The establishment of proper cell interaction is whether it fundamentally results in proper cell adhesion, migration and movement. For the realization of the multiplex membrane tool, a Poly(D,Llactide-co-glycolide)(PLGA) membrane was selected, whose well-defined morphological, physico-chemical and mechanical properties are adequate for supporting and boosting neuronal survival and differentiation [15]. Indeed, a previous study already showed that this membrane represents a highly cell-compatible matrix where neurons can properly organize according to a complex network with functioning cell-cell contacts and interaction.…”

Hollow Fiber and Nanofiber Membranes in Bioartificial Liver and Neuronal Tissue Engineering

“…In this context, decades of advances in membrane technology have led to its application in neuroscience and neural engineering. Several papers indicated that membrane-based systems provide in vitro advanced devices that enhance neuronal growth and differentiation and enable the repair and regeneration of the nervous system [De Bartolo et al, 2008;Morelli et al, 2010Morelli et al, , 2015bMorelli et al, , 2017a. In addition, neuronal biohybrid membrane systems can mimic specific features of the in vivo neuronal environment and, therefore, are also widely used as in vitro brain tissue models for pharmacological screening and as investigational platform for neurodegenerative diseases [Giusi et al, 2009;Morelli et al, 2014Morelli et al, , 2016Morelli et al, , 2019Morelli et al, , 2021Piscioneri et al, 2015Piscioneri et al, , 2021Mele et al, 2017].…”