In live organisms, cells are embedded in tissue-specific extracellular matrix (ECM), which provides chemical and mechanical signals important for cell differentiation, migration, and overall functionality. Careful reproduction of ECM properties in artificial cell scaffolds is necessary to get physiologically relevant results of in vitro studies and produce robust materials for cell and tissue engineering. Nanoarchitectonics is a contemporary way to building complex materials from nano-scale objects of artificial and biological origin. Decellularized ECM (dECM), remaining after cell elimination from organs, tissues and cell cultures is arguably the closest equivalent of native ECM achievable today. dECM-based materials can be used as templates or components for producing cell scaffolds using nanoarchitectonic approach. Irrespective of the form, in which dECM is used (whole acellular organ/tissue, bioink or hydrogel), the local stiffness of the dECM scaffold must be evaluated, since the fate of seeded cells depends on the mechanical properties of their environment. Careful dECM characterization is also necessary to reproduce essential ECM traits in artificial cell scaffolds by nanoparticle assembly. Atomic force microscopy (AFM) is a valuable characterization tool, as it allows simultaneous assessment of mechanical and topographic features of the scaffold, and additionally evaluate the efficiency of decellularization process and preservation of the extracellular matrix. This review depicts the current application of AFM in the field of dECM-based materials, including the basics of AFM technique and the use of flicker-noise spectroscopy (FNS) method for the quantification of the dECM micro- and nanostructure.

The growing applications of tissue engineering technologies warrant the search and development of biocompatible materials with an appropriate strength and elastic moduli. Here, we have extensively studied a collagenous membrane (GSCM) separated from the mantle of the Giant squid Dosidicus Gigas in order to test its potential applicability in regenerative medicine. To establish the composition and structure of the studied material, we analyzed the GSCM by a variety of techniques, including amino acid analysis, SDS-PAGE, and FTIR. It has been shown that collagen is a main component of the GSCM. The morphology study by different microscopic techniques from nano- to microscale revealed a peculiar packing of collagen fibers forming laminae oriented at 60–90 degrees in respect to each other, which, in turn, formed layers with the thickness of several microns (a basketweave motif). The macro- and micromechanical studies showed high values of the Young’s modulus and tensile strength. No significant cytotoxicity of the studied material was found by the cytotoxicity assay. Thus, the GSCM consists of a reinforced collagen network, has high mechanical characteristics, and is non-toxic, which makes it a good candidate for the creation of a scaffold material for tissue engineering.

In the realm of scaffold-free cell therapies, there is a questto develop organotypic three-dimensional (3D) tissue surrogates in vitro, capitalizing on the inherent ability of cells to create tissues with an efficiency and sophistication that still remains unmatched by human-made devices. In this study, we explored the properties of scaffolds obtained by the electrospinning of a thermosensitive copolymer, poly(N-isopropylacrylamide-co-N-tert-butylacrylamide) (P(NIPAM-co-NtBA)), intended for use in such therapies. Two copolymers with molecular weights of 123 and 137 kDa and a content of N-tert-butylacrylamide of ca. 15 mol% were utilized to generate 3D scaffolds via electrospinning. We examined the morphology, solution viscosity, porosity, and thickness of the spun matrices as well as the mechanical properties and hydrophobic–hydrophilic characteristics of the scaffolds. Particular attention was paid to studying the influence of the thermosensitive polymer’s molecular weight and dispersity on the resultant scaffolds’ properties and the role of electroforming parameters on the morphology and mechanical characteristics of the scaffolds. The cytotoxicity of the copolymers and interaction of cells with the scaffolds were also studied. Our findings provide significant insight into approaches to optimizing scaffolds for specific cell cultures, thereby offering new opportunities for scaffold-free cell therapies.