Mutual influence of selenium nanoparticles and FGF2-STAB® on biocompatible properties of collagen/chitosan 3D scaffolds: in vitro and ex ovo evaluation

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.

Bioactive Collagen Hydrolysate-Chitosan/Essential Oil Electrospun Nanofibers Designed for Medical Wound Dressings

“…Instead, the electrospun sample based on collagen hydrolysate extracted from rabbit skin mixed with chitosan (coded HCR) was very efficient against all the tested samples, excepting E. coli. A high inhibition effect towards S. aureus was also reported in the case of collagen/chitosan scaffolds [58].…”

Lumbar Interbody Fusion Conducted on a Porcine Model with a Bioresorbable Ceramic/Biopolymer Hybrid Implant Enriched with Hyperstable Fibroblast Growth Factor 2

“…Newly, we verified the safety, healing, and angiogenic potential of collagenbased scaffolds enriched with FGF2-STAB ® as full skin replacement in vivo on 3-month-old White New Zealand rabbits [29]. Biodegradable scaffolds revealed promising results in cell culture experiments and have displayed high suitability and biocompatibility to be used as a transferable scaffold for tissue reconstruction [30]. However, the aforementioned collagen-based scaffolds are very soft and elastic, being excellent for skin regeneration but lack structural stability upon hydration and mechanical strength, which limit their applications in particular bone tissues [31].…”

“…Therefore, in the present study, we used the same BCP 3D matrix as a control group for 21 days cytotoxicity evaluation of the BCP matrix impregnated with collagen, oxidized cellulose, and FGF2-STAB ® (BHI). The amount of FGF2-STAB ® protein has been widely evaluated in our previous studies [29,30].…”

Enterococcus spp. Cell-Free Extract: An Abiotic Route for Synthesis of Selenium Nanoparticles (SeNPs), Their Characterisation and Inhibition of Escherichia coli

“…The mechanism of the cytotoxicity of SeNPs against bacteria are still unclear. However, it is known that many bacteria have a negatively charged surface and the zeta potential of the nanoparticles plays an important role due to this [44]. In this study, the gram-negative bacteria E. coli was used.…”