Abstract
Biomimetic materials are designed to stimulate specific cellular responses at the molecular level. To improve the soundness of in vitro testing of the biological impact of new materials, appropriate cell systems and technologies must be standardized also taking regulatory issues into consideration. In this study, the biological and molecular effects of different scaffolds on three neural systems, that is, the neural cell line SH-SY5Y, primary cortical neurons, and neural stem cells, were compared. The effect of poly(L-lactic acid) scaffolds having different surface geometry (conventional two-dimensional seeding flat surface, random or aligned fibers as semi3D structure) and chemical functionalization (laminin or ECM extract) were studied. The endpoints were defined for efficacy (i.e., neural differentiation and neurite elongation) and for safety (i.e., cell death/survival) using high-content analysis. It is demonstrated that (i) the definition of the biological properties of biomaterials is profoundly influenced by the test system used; (ii) the definition of the in vitro safety profile of biomaterials for neural repair is also influenced by the test system; (iii) cell-based high-content screening may well be successfully used to characterize both the efficacy and safety of novel biomaterials, thus speeding up and improving the soundness of this critical step in material science having medical applications.
Highlights
The potential applications of tissue engineering products for regenerative medicine in acute and chronic diseases of the nervous system are quite wide
In this work we compared the biological and molecular effects of different materials on three cell systems commonly used for these purposes, that is, the neural cell line SH-SY5Y, primary cortical neurons, and neural stem/precursor cells (NSC)
The following variables were included in the study: (A) materials and topography of cell substrates: conventional glass/plastic 2D, semi three-dimensional random electrospun poly(L-lactic acid) (PLLA) scaffolds and semi three- dimensional aligned electrospun PLLA scaffolds; (B) substrate chemical functionalization: no-coating, laminin coating of the above materials and whole ECM extract coating of the above materials; (C) neural cell type: SH-SY5Y neural differentiated cell line, primary cortical neurons, and neural stem cells; (D) assay and technologies for toxicology testing: conventional low-throughput and cell-based high throughput screening
Summary
The potential applications of tissue engineering products for regenerative medicine in acute and chronic diseases of the nervous system are quite wide. There is an extensive literature aimed to compare the biological impact of different scaffolds for neural repair, considering the chemical composition of the materials, architecture, functionalization, etc.[1] these studies often brought to contradictory results when translated in animals, and this could be at least partially due to the different cell systems and readout technologies used.[2] key biological differences in the cellular test systems (i.e., cellular composition, maturation time, culture media composition, growth factor production, etc.) make it very hard to compare results and judge the ‘‘biomaterial effect on neural cells’’.3,4. In this work we compared the biological and molecular effects of different materials on three cell systems commonly used for these purposes, that is, the neural cell line SH-SY5Y, primary cortical neurons, and neural stem/precursor cells (NSC). We investigated glass substrates and poly(L-lactic acid) (PLLA) scaffolds having different surface geometry (conventional twodimensional seeding [2D] flat surface, random or aligned fibers as semi3D structure) and chemical functionalization (laminin or ECM extract)
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