Abstract
Several types of promising cell-based therapies for tissue regeneration have been developing worldwide. However, for successful therapeutical application of cells in this field, appropriate scaffolds are also required. Recently, the research for suitable scaffolds has been focusing on polymer hydrogels due to their similarity to the extracellular matrix. The main limitation regarding amino acid-based hydrogels is their difficult and expensive preparation, which can be avoided by using poly(aspartamide) (PASP)-based hydrogels. PASP-based materials can be chemically modified with various bioactive molecules for the final application purpose. In this study, dopamine containing PASP-based scaffolds is investigated, since dopamine influences several cell biological processes, such as adhesion, migration, proliferation, and differentiation, according to the literature. Periodontal ligament cells (PDLCs) of neuroectodermal origin and SH-SY5Y neuroblastoma cell line were used for the in vitro experiments. The chemical structure of the polymers and hydrogels was proved by 1H-NMR and FTIR spectroscopy. Scanning electron microscopical (SEM) images confirmed the suitable pore size range of the hydrogels for cell migration. Cell viability assay was carried out according to a standardized protocol using the WST-1 reagent. To visualize three-dimensional cell distribution in the hydrogel matrix, two-photon microscopy was used. According to our results, dopamine containing PASP gels can facilitate vertical cell penetration from the top of the hydrogel in the depth of around 4 cell layers (~150 μm). To quantify these observations, a detailed image analysis process was developed and firstly introduced in this paper.
Highlights
IntroductionTissue engineering is a rapidly developing and promising field that applies living cells growing on complex biocompatible matrices called scaffolds, to create tissue-like structures [1,2,3]
We have demonstrated that thiol groups in the PASP hydrogels facilitate cell adhesion and proliferation [15]
Since the preparation of the hydrogels consists of several steps
Summary
Tissue engineering is a rapidly developing and promising field that applies living cells growing on complex biocompatible matrices called scaffolds, to create tissue-like structures [1,2,3]. These scaffolds can be used for medical therapies repairing the injured or diseased area of tissues [1,2,3], but they are suitable for testing the efficacy and toxicity of various drugs [2,4]. A high number of research projects related to tissue engineering are focusing on hydrogels, due to their structural similarity to the natural environment of the living cells, i.e., the extracellular matrix (ECM). The adequate behavior of the cells can be achieved by incorporating growth or differentiation factors [6], as well as other bioactive agents like proteins, peptides, or small molecules into the hydrogel matrix [1,3,7,8]
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