Abstract
In a biological system, nanoparticles (NPs) may interact with biomolecules. Specifically, the adsorption of proteins on the nanoparticle surface may influence both the nanoparticles' and proteins' overall bio-reactivity. Nevertheless, our knowledge of the biocompatibility and risk of exposure to nanomaterials is limited. Here, in vitro and ex ovo biocompatibility of naturally based crosslinked freeze-dried 3D porous collagen/chitosan scaffolds, modified with thermostable fibroblast growth factor 2 (FGF2-STAB®), to enhance healing and selenium nanoparticles (SeNPs) to provide antibacterial activity, were evaluated. Biocompatibility and cytotoxicity were tested in vitro using normal human dermal fibroblasts (NHDF) with scaffolds and SeNPs and FGF2-STAB® solutions. Metabolic activity assays indicated an antagonistic effect of SeNPs and FGF2-STAB® at high concentrations of SeNPs. The half-maximal inhibitory concentration (IC50) of SeNPs for NHDF was 18.9 µg/ml and IC80 was 5.6 µg/ml. The angiogenic properties of the scaffolds were monitored ex ovo using a chick chorioallantoic membrane (CAM) assay and the cytotoxicity of SeNPs over IC80 value was confirmed. Furthermore, the positive effect of FGF2-STAB® at very low concentrations (0.01 µg/ml) on NHDF metabolic activity was observed. Based on detailed in vitro testing, the optimal concentrations of additives in the scaffolds were determined, specifically 1 µg/ml of FGF2-STAB® and 1 µg/ml of SeNPs. The scaffolds were further subjected to antimicrobial tests, where an increase in selenium concentration in the collagen/chitosan scaffolds increased the antibacterial activity. This work highlights the antimicrobial ability and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB® and SeNPs. Moreover, we suggest that these sponges could be used as scaffolds for growing cells in systems with low mechanical loading in tissue engineering, especially in dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration. Due to their antimicrobial properties, these scaffolds are also highly promising for tissue replacement requiring the prevention of infection.
Highlights
Hydrogels have become increasingly studied as matrices for tissue engineering [25]
Biocompatibility using human dermal fibroblasts cell line Biocompatibility of scaffolds According to our previous studies [6, 9, 19] the FGF2STAB® concentration of 2 μg/ml and the selenium nanoparticles (SeNPs) concentrations of 2, 10 and 20 μg/ml were selected for Collagen/ Chitosan scaffold preparation
In this study, Collagen/Chitosan scaffolds were enriched with selenium nanoparticles (SeNPs) and hyperstable fibroblast growth factor 2 (FGF2-STAB®) to develop safe antibacterial and vasculogenic scaffolds for possible aplication as tissue-engineered skin replacement
Summary
Hydrogels have become increasingly studied as matrices for tissue engineering [25]. Derived hydrogelforming polymers have frequently been used in tissue engineering applications because they are either components of, or have macromolecular properties, similar to the natural extracellular matrix (ECM) [10]. The. Muchová et al J Nanobiotechnol (2021) 19:103 disadvantage of natural material hydrogels is its poor antimicrobial activity, making it a good substrate for bacterial growth and leading to severe infections and inflammation [16]. Muchová et al J Nanobiotechnol (2021) 19:103 disadvantage of natural material hydrogels is its poor antimicrobial activity, making it a good substrate for bacterial growth and leading to severe infections and inflammation [16] This characteristic has led to the need to modify natural material hydrogels to incorporate antibacterial properties to prevent these undesirable side reactions. Metal and metal oxide nanoparticles, well known for their highly potent antibacterial effects [27] include silver (Ag), iron oxide (Fe3O4), titanium oxide (TiO2), copper oxide (CuO), and zinc oxide (ZnO)
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